CN101396361B - Medicine composition containing L-carnosine for suspending the development of the cataract - Google Patents

Medicine composition containing L-carnosine for suspending the development of the cataract Download PDF

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CN101396361B
CN101396361B CN 200710161601 CN200710161601A CN101396361B CN 101396361 B CN101396361 B CN 101396361B CN 200710161601 CN200710161601 CN 200710161601 CN 200710161601 A CN200710161601 A CN 200710161601A CN 101396361 B CN101396361 B CN 101396361B
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carnosine
catalin
compositions
crystalline lens
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CN101396361A (en
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刘继东
杨宇春
孙洋
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Shenyang Sinqi Pharmaceutical Co Ltd
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Shenyang Sinqi Pharmaceutical Co Ltd
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Abstract

The invention relates to a medical combination containing L-carnosine, in particular to an ophthalmic gel preparation containing the L-carnosine and the application of the medical combination and the ophthalmic gel preparation in the treatment of the cataract and preventing or retarding the development of the cataract.

Description

Be used for delaying the pharmaceutical composition that contains L-Carnosine of development of cataracts
Technical field
The present invention relates to a kind of Pharmaceutical composition that contains L-Carnosine, particularly a kind of eye-gel preparation that contains L-Carnosine, and described pharmaceutical composition and eye-gel preparation are used for cataractous treatment, prevent or delay the purposes of cataractous progress.
Background technology
The treatment of cataract disease is the social problem of being badly in need of solution, and cataract is the first diseases causing blindness in the world, although surgical operation can improve patient's vision, operation risk, serious post-operative complication and the economic problems that cause therefrom still can not be ignored.So the various countries scholar is devoted to seek effective anti-cataract medicine always.
At present; for screening has prevention, treatment or suppresses the candidate compound that cataract develops; general isolated rat cataract model by hydrogen peroxide-induced; investigate testing compound to the protective effect of Oxidative Damage of Lens, thereby indirectly judge described testing compound for prevention, treatment or suppress the effect that cataract develops [1]
Studies have shown that peptides can increase the depleted and aging ability of cell resistance [2]Peptides has the effect of defying age and prevention age related disease.Oxidative stress, radical damage are the reasons that causes aging and age related disease.The unsaturated fatty acid effect of free radical and cell membrane makes lipid peroxidation produce conjugation alkene class and end-product malonaldehyde (MDA).Affect the permeability of film, infringement Na pump, Ca pump, Ion imbalance: MDA also generates the conjugate with fluorescence Schif residue with the protein active group, causes protein to build up lenticular opacity.
L-Carnosine can protect the d-crystallin not oxidated stress damage, have and remove free radical and lipid peroxy chemical compound, the cataract development can be treated or suppress to the effect of protection Na-K pump ATP enzyme [3]
L-Carnosine is the dipeptides that is made of Beta-alanine and L-Histidine, extensively is present in each histoorgan of body, and particularly in muscular tissue, brain and crystalline lens, concentration can reach about 20mmol/L.L-Carnosine is the nontoxic medicine of endogenous, but L-Carnosine Scavenger of ROS free radical not only, and can repair the enzyme system of being responsible for the Scavenger of ROS free radical, increase the activity of surface of cell membrane enzyme.
L-Carnosine is at present multiplex in laboratory research, there is no the launch that clearly is used for delaying the development of cataracts aspect as opthalmological.
A kind of flushing liquor of protecting cornea is disclosed in Russian Federation's patent application (RU2114587), for refraction correction surgery and ophthalmology internal cavity operation.Contain sodium chloride, sodium dihydrogen phosphate, sodium hydrogen phosphate, Portugal's ammonia polysaccharide, cellulose derivative (alkyl, carboxyalkyl or hydroxyalkyl cellulose derivative) and pure water in the described flushing liquor.Especially, the optional L-Carnosine that contains 0.5-20.0g/l also in the washing liquid of described protection cornea.In described medical composite for eye, L-Carnosine is used for reducing the rear microcosmic edema of operation as biomembrane water layer and lipid antioxidant and protective agent mutually.
Russian Federation's patent application (RU2115413) also discloses a kind of Ophthalmologic irrigation solutions, can be used for substituting when ophthalmologic operation is got involved camera oculi anterior body fluid, flushing camera oculi anterior and its hetero-organization.Contain sodium chloride, sodium dihydrogen phosphate, sodium hydrogen phosphate, carnosine and pure water in this flushing liquor, and optional glucose and cellulose derivative.Concrete, also contain the L-Carnosine of 0.5-20.0g/l in the described flushing liquor, be used for corneal edema and reduce.
Russian Federation's patent application (RU2121324) also discloses a kind of Ophthalmologic irrigation solutions for ophthalmology inner chamber and the intervention of non-internal cavity operation, and is better than the counterbalance effect of existing flushing liquor.Contain sodium chloride, sodium dihydrogen phosphate, sodium hydrogen phosphate, water and Portugal's amine polysaccharide, L-Carnosine, cytochrome C in this flushing liquor.Concrete, the content of L-Carnosine is 0.002-0.2% in the described flushing liquor, is used for stablizing taking a tonic or nourishing food to build up one's health of cornea endothelial cell membrane and appropriateness.
Also disclose a kind of carnosine eye drops in Russian Federation's patent application (RU2201213), it contains the carnosine of physiological salt solution, borate buffer solution and 0.01-2.0 % by weight.Choose wantonly, described eye drop also can contain thickening agent and/or antiseptic.Wherein carnosine is used for stabilizing cell membrane as antioxidant, promotes damaged cell to recover, and makes damaged tissue metabolism process normal.
Also disclose a kind of in Russian Federation's patent application (RU2288702) and used washing liquid, be used for cataract extraction ophthalmologic operation, the filling of various ophthalmologic operation, wherein contained sodium chloride, sodium dihydrogen phosphate, sodium hydrogen phosphate, carnosine, Portugal's amine polysaccharide and water.Wherein said carnosine act as antioxidant, and certain antiinflammatory action is arranged.
Utilize the isolated rat cataract model of hydrogen peroxide-induced, the inventor finds, traditionally always eye with in the flushing liquor as the L-Carnosine of antioxidant and antiinflammatory, it has obvious anti-Oxidative Damage of Lens effect in the finite concentration scope, can obviously improve the activity of crystalline lens antioxidase, delay cataractous progress.
Summary of the invention
One aspect of the present invention relates to a kind of pharmaceutical composition be used to delaying development of cataracts, and it contains L-Carnosine, buffer salt, osmotic pressure regulator.According to the dosage form of described pharmaceutical composition, optional antiseptic and/or the thickening agent of containing also in the described pharmaceutical composition.
Its pH scope of pharmaceutical composition of the present invention preferably remains on 6.8-8.0.Remain on 6.8-8.0 in order to keep pharmaceutical composition pH of the present invention, adoptable buffer salt is selected from boric acid-Borax, sodium hydrogen phosphate-sodium dihydrogen phosphate, boric acid-sodium carbonate and boric acid-sodium acetate.
In the pharmaceutical composition of the present invention, preferably its osmotic pressure remains on 260-340mosm/L.In order to keep the osmotic pressure of pharmaceutical composition of the present invention, adoptable osmotic pressure regulator is selected from sodium chloride, potassium chloride, glycerol, glucose, propylene glycol and mannitol.
The antiseptic that is applicable in the pharmaceutical composition of the present invention is selected from: ethyl hydroxybenzoate, chlorhexidine gluconate, Chlorhexidine hydrochloride, chlorhexidine acetate, cetrimonium bromide, benzalkonium chloride, benzalkonium bromide, phenoxyethanol and chlorobutanol.
The thickening agent that is applicable in the pharmaceutical composition of the present invention is selected from: hyaluronic acid sodium, polyvinyl alcohol, polyvidone and water-soluble chitosan.
In order to realize that L-Carnosine of the present invention delays the function of development of cataracts, (by weight percentage) L-Carnosine is 0.5-10.0% in the described pharmaceutical composition.When containing antiseptic or thickening agent in the pharmaceutical composition, antiseptic is 0.001-5% (by weight percentage), and thickening agent is 0.01-3% (by weight percentage).
Further preferred, (by weight percentage) L-Carnosine is 1.0-6.0% in the pharmaceutical composition of the present invention.
When antiseptic was selected ethyl hydroxybenzoate in the pharmaceutical composition of the present invention, further preferred its consumption (by weight percentage) was 0.01%-0.09%; When antiseptic in the pharmaceutical composition of the present invention was selected a kind of in chlorhexidine gluconate, Chlorhexidine hydrochloride, the chlorhexidine acetate, further preferred its consumption (by weight percentage) was 0.001%-0.01%; When antiseptic was selected cetrimonium bromide in the pharmaceutical composition of the present invention, further preferred its consumption (by weight percentage) was 0.01%-0.09%; When antiseptic was selected benzalkonium chloride or benzalkonium bromide in the pharmaceutical composition of the present invention, further preferred its consumption (by weight percentage) was 0.002%-0.03%; When antiseptic was selected phenoxyethanol in the pharmaceutical composition of the present invention, further preferred its consumption (by weight percentage) was 1.0%-5.0%; When antiseptic was selected chlorobutanol in the pharmaceutical composition of the present invention, further preferred its consumption (by weight percentage) was 0.1%-0.9%.
When thickening agent was selected hyaluronic acid sodium in the pharmaceutical composition of the present invention, further preferred its consumption (by weight percentage) was 0.01%-0.1%; When thickening agent was selected polyvinyl alcohol in the pharmaceutical composition of the present invention, further preferred its consumption (by weight percentage) was 0.1%-2.8%; When thickening agent was selected polyvidone in the pharmaceutical composition of the present invention, further preferred its consumption (by weight percentage) was 0.5%-2%; When thickening agent was selected water-soluble chitosan in the pharmaceutical composition of the present invention, further preferred its consumption (by weight percentage) was 0.1%-2.5%.
Isolated rat cataract model by hydrogen peroxide-induced confirms that the left-handed polypeptide in the finite concentration scope has obvious anti-Oxidative Damage of Lens effect, can obviously improve the activity of crystalline lens antioxidase, delays cataractous progress.
Concrete, in embodiments of the invention, the preferred concentration range of described L-Carnosine is counted 0.5-10.0% (weight) by the weight of total pharmaceutical composition.
Concrete, the acting body that delays cataractous progress includes but not limited to following aspect now: reduce or postpone lenticular muddy degree, keep or keep the content of water-solubility protein in the crystalline lens, and the activity that keeps or keep SOD in the crystalline lens, thereby can obviously improve the aspects such as activity of crystalline lens antioxidase.
In the present invention, described pharmaceutical composition be used to delaying development of cataracts can be arbitrary dosing eyes dosage form that is suitable for cataract therapy, includes but not limited to: eye drop, Eye ointments, ocular inserts, gel, situ-gel drug-supplying system, colloidal drug delivery system (comprising microemulsion, liposome, nanoparticle and nanocapsule), microsphere, implant.
Concrete, eye drop (eye drop) refers to contain the solution of L-Carnosine.Those of ordinary skills know, and in order to improve the bioavailability of eye drop medicine, can add suitable pharmaceutic adjuvant according to concrete demand in prescription forms.
For example, be stranded in for a long time ophthalmic in order to make medicine, reduce the medicinal liquid loss, can in eye drop, add thickening agent, include but not limited to hyaluronic acid sodium, polyvinyl alcohol, polyvidone and water-soluble chitosan etc.
Eye ointments (eye ointments) is the sterilized ointment for eye, and such as can selecting vaseline and lanoline etc. to mix oleaginous base, or the collagen with suitable stickiness and denseness of 1%-5% is as ointment base.
Ocular inserts (eye pellicles) is with medicine dissolution or is dispersed in the thin film formulations that is processed in the suitable filmogen.The filmogen of ocular inserts comprises for example polyvinyl alcohol (PVA), or the mixture of PVA and natural macromolecular material Bletilla glucomannan.
The situ-gel drug-supplying system contains the polymer that can occur to change mutually in forming, and with the form eye dripping of liquid phase, pleasing to the eye rear horse back changes gel phase into.Polymer during situ-gel drug-supplying system commonly used forms comprises for example poloxamer407, tetronics, CAP (cap), lactex, gelritetm and polysaccharide.
Be suitable for the form that medicine that L-Carnosine is used for delaying development of cataracts also can be colloidal drug delivery system that contains of the present invention, comprise the forms such as microemulsion, liposome, nanoparticle and nanocapsule.
In addition, be suitable for of the present inventionly containing medicine that L-Carnosine is used for delaying development of cataracts and for example also can adopting particle diameter the tiny of 1-10 μ m and microsphere or the implant form of uniform matrix type particle form.
Concrete, the prescription that is suitable for delaying the pharmaceutical composition of the present invention of development of cataracts includes but not limited to following example:
Composite formula 1
Contain L-Carnosine 5g in each kilogram of composition, ethyl hydroxybenzoate 0.3g, hyaluronic acid sodium 0.5g, control pH value with boric acid-Borax between 6.8 to 8.0 as buffer salt, regulate osmotic pressure to 260-340mosm/L with an amount of sodium chloride, surplus is pure water, is prepared into according to a conventional method eye drop.
Composite formula 2
Contain L-Carnosine 10g in each kilogram of composition, chlorhexidine gluconate 0.05g, polyvinyl alcohol 10g, control pH value with sodium hydrogen phosphate-sodium dihydrogen phosphate between 6.8 to 8.0 as buffer salt, regulate osmotic pressure to 260-340mosm/L with an amount of glycerol, surplus is pure water, is prepared into according to a conventional method eye drop.
Composite formula 3
Contain L-Carnosine 50g in each kilogram of composition, phenoxyethanol 50g, polyvidone 10g, control pH value with boric acid-sodium carbonate between 6.8 to 8.0 as buffer salt, regulate osmotic pressure to 260-340mosm/L with an amount of potassium chloride, surplus is pure water, is prepared into according to a conventional method eye drop.
Composite formula 4
Contain L-Carnosine 80g in each kilogram of composition, cetrimonium bromide 0.1g, hyaluronic acid sodium 5g, control pH value with boric acid-sodium acetate between 6.8 to 8.0 as buffer salt, regulate osmotic pressure to 260-340mosm/L with an amount of potassium chloride, surplus is pure water, is prepared into according to a conventional method eye drop.
Composite formula 5
Contain L-Carnosine 100g in each kilogram of composition, chlorobutanol 3g, water-soluble chitosan 6g, control pH value with boric acid-Borax between 6.8 to 8.0 as buffer salt, regulate osmotic pressure to 260-340mosm/L with an amount of glucose, surplus is pure water, is prepared into according to a conventional method eye drop.
Composite formula 6
Contain L-Carnosine 60g in each kilogram of composition, ethyl hydroxybenzoate 0.3g, hyaluronic acid sodium 1g, control pH value with boric acid-sodium acetate between 6.8 to 8.0 as buffer salt, regulate osmotic pressure to 260-340mosm/L with an amount of propylene glycol, surplus is pure water, is prepared into according to a conventional method eye drop.
Composite formula 7
Contain L-Carnosine 50g in each kilogram of composition, control pH value with sodium hydrogen phosphate-sodium dihydrogen phosphate between 6.8 to 8.0 as buffer salt, regulate osmotic pressure to 260-340mosm/L with an amount of glycerol, surplus is pure water, is prepared into the single dose eye drop with auto-filling equipment.
Composite formula 8
Contain L-Carnosine 30g in each kilogram of composition, control pH value with boric acid-Borax between 6.8 to 8.0 as buffer salt, regulate osmotic pressure to 260-340mosm/L with an amount of sodium chloride, surplus is pure water, is prepared into the single dose eye drop with auto-filling equipment.
The invention still further relates to a kind of eye-gel preparation be used to delaying development of cataracts, it contains L-Carnosine, buffer salt, osmotic pressure regulator, gel-type vehicle and antiseptic.According to the concrete dosage form of described eye-gel preparation, the optional thickening agent that contains also in the described preparation.
Its pH scope of eye-gel preparation that contains L-Carnosine of the present invention preferably remains on 6.8-8.0.The eye-gel preparation pH that contains L-Carnosine in order to keep the present invention remains on 6.8-8.0, and adoptable buffer salt is selected from boric acid-Borax, sodium hydrogen phosphate-sodium dihydrogen phosphate, boric acid-sodium carbonate and boric acid-sodium acetate.
In the eye-gel preparation that contains L-Carnosine of the present invention, preferably its osmotic pressure remains on 260-340mosm/L.The osmotic pressure that contains the eye-gel preparation of L-Carnosine in order to keep the present invention, adoptable osmotic pressure regulator is selected from sodium chloride, potassium chloride, glycerol, glucose, propylene glycol and mannitol.
Be applicable to the gel-type vehicle that contains the eye-gel preparation of L-Carnosine of the present invention and be selected from water-soluble cellulose, carbomer, poloxamer and water-soluble chitosan.Water-soluble cellulose comprises: sodium carboxymethyl cellulose, hydroxyethyl-cellulose, hydroxyl second methylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, methylcellulose etc.The model of carbomer mainly contains 910,934,934P, 940,941,971P, 974,974P, 980,981,1342.The model of poloxamer mainly comprises: 124,188,237,338,407.Water-soluble chitosan mainly refers to have the water-soluble chitosan derivants such as the water-soluble chitosan of certain deacetylation and carboxymethyl chitosan.
The antiseptic that is applicable in the eye-gel preparation that contains L-Carnosine of the present invention is selected from: ethyl hydroxybenzoate, chlorhexidine gluconate, Chlorhexidine hydrochloride, chlorhexidine acetate, cetrimonium bromide, benzalkonium chloride, benzalkonium bromide, phenoxyethanol and chlorobutanol.
The thickening agent that is applicable in the gel preparation that contains L-Carnosine of the present invention is selected from: hyaluronic acid sodium, polyvinyl alcohol, polyvidone and water-soluble chitosan.
In order to realize that L-Carnosine of the present invention delays the function of development of cataracts, (by weight percentage) L-Carnosine is 0.5-10.0% in the described eye-gel preparation; Gel-type vehicle is 0.05-30%, and antiseptic is 0.001-5%, and thickening agent is 0.01-3%.
Further preferred, (by weight percentage) L-Carnosine is 1.0-6.0% in the eye-gel preparation of the present invention.
When gel-type vehicle was selected water-soluble cellulose in the eye-gel preparation of the present invention, further preferred its consumption (by weight percentage) was 0.5%-6%; When gel-type vehicle was selected carbomer in the eye-gel preparation of the present invention, further preferred its consumption (by weight percentage) was 0.1%-0.9%; When gel-type vehicle was selected poloxamer in the eye-gel preparation of the present invention, further preferred its consumption (by weight percentage) was 12%-27%; When gel-type vehicle was selected water-soluble chitosan in the eye-gel preparation of the present invention, further preferred its consumption (by weight percentage) was 0.5%-5%.
When antiseptic was selected ethyl hydroxybenzoate in the eye-gel preparation of the present invention, further preferred its consumption (by weight percentage) was 0.01%-0.09%; When antiseptic in the eye-gel preparation of the present invention was selected a kind of in chlorhexidine gluconate, Chlorhexidine hydrochloride, the chlorhexidine acetate, further preferred its consumption (by weight percentage) was 0.001%-0.01%; When antiseptic was selected cetrimonium bromide in the eye-gel preparation of the present invention, further preferred its consumption (by weight percentage) was 0.01%-0.09%; When antiseptic was selected benzalkonium chloride or benzalkonium bromide in the pharmaceutical composition of the present invention, further preferred its consumption (by weight percentage) was 0.002%-0.03%; When antiseptic was selected phenoxyethanol in the eye-gel preparation of the present invention, further preferred its consumption (by weight percentage) was 1.0%-5.0%; When antiseptic was selected chlorobutanol in the eye-gel preparation of the present invention, further preferred its consumption (by weight percentage) was 0.1%-0.9%.
When thickening agent was selected hyaluronic acid sodium in the eye-gel preparation of the present invention, further preferred its consumption (by weight percentage) was 0.01%-0.1%; When thickening agent was selected polyvinyl alcohol in the eye-gel preparation of the present invention, further preferred its consumption (by weight percentage) was 0.1%-2.8%; When thickening agent was selected polyvidone in the eye-gel preparation of the present invention, further preferred its consumption (by weight percentage) was 0.5%-2%; When thickening agent was selected water-soluble chitosan in the eye-gel preparation of the present invention, further preferred its consumption (by weight percentage) was 0.1%-2.5%.
Concrete, the prescription that contains the eye-gel preparation of L-Carnosine of the present invention that is suitable for delaying development of cataracts includes but not limited to following example:
Eye-gel preparation prescription 1
Contain L-Carnosine 5g in each kilogram eye-gel preparation, sodium carboxymethyl cellulose 40g, ethyl hydroxybenzoate 0.3g, hyaluronic acid sodium 0.1g, control pH value with boric acid-Borax between 6.8 to 8.0 as buffer salt, regulate osmotic pressure to 260-340mosm/L with an amount of propylene glycol, surplus is pure water, is prepared into according to a conventional method gel for eye use.
Eye-gel preparation prescription 2
Contain L-Carnosine 10g in each kilogram eye-gel preparation, hydroxypropyl methylcellulose 50g, chlorhexidine gluconate 0.05g, polyvinyl alcohol 5g, control pH value with boric acid-sodium acetate between 6.8 to 8.0 as buffer salt, regulate osmotic pressure to 260-340mosm/L with an amount of glycerol, surplus is pure water, is prepared into according to a conventional method gel for eye use.
Eye-gel preparation prescription 3
Contain L-Carnosine 50g in each kilogram eye-gel preparation, carbomer 934 is 24g, phenoxyethanol 50g, polyvidone 2g, control pH value with sodium hydrogen phosphate-sodium dihydrogen phosphate between 6.8 to 8.0 as buffer salt, regulate osmotic pressure to 260-340mosm/L with an amount of mannitol, surplus is pure water, is prepared into according to a conventional method gel for eye use.
Eye-gel preparation prescription 4
Contain L-Carnosine 80g in each kilogram eye-gel preparation, Carbopol is 18g, chlorhexidine acetate 0.05g, water-soluble chitosan 0.5g, control pH value with boric acid-sodium acetate between 6.8 to 8.0 as buffer salt, regulate osmotic pressure to 260-340mosm/L with an amount of propylene glycol, surplus is pure water, is prepared into according to a conventional method gel for eye use.
Eye-gel preparation prescription 5
Contain L-Carnosine 100g in each kilogram eye-gel preparation, water-soluble chitosan 35g, chlorobutanol 7g, hyaluronic acid sodium 0.2g, control pH value with boric acid-Borax between 6.8 to 8.0 as buffer salt, regulate osmotic pressure to 260-340mosm/L with an amount of glucose, surplus is pure water, is prepared into according to a conventional method gel for eye use.
Eye-gel preparation prescription 6
Contain L-Carnosine 60g in each kilogram eye-gel preparation, poloxamer188 is 180g, cetrimonium bromide 0.2g, hyaluronic acid sodium 1g, control pH value with boric acid-sodium acetate between 6.8 to 8.0 as buffer salt, regulate osmotic pressure to 260-340mosm/L with an amount of propylene glycol, surplus is pure water, is prepared into according to a conventional method gel for eye use.
Eye-gel preparation prescription 7
Contain L-Carnosine 10g in each kilogram eye-gel preparation, hydroxypropyl methylcellulose 50g, chlorobutanol 7g, control pH value with boric acid-sodium carbonate between 6.8 to 8.0 as buffer salt, regulate osmotic pressure to 260-340mosm/L with an amount of glycerol, surplus is pure water, is prepared into according to a conventional method gel for eye use.
Below, in conjunction with specific embodiments the present invention's pharmaceutical composition required for protection and eye-gel preparation further are illustrated.
Embodiment
The external test of pesticide effectiveness of embodiment 1 L-Carnosine
Experiment material
Instrument and equipment: superclean bench (AVC-5A1, Singapore ESCO company product), CO2 gas incubator (Innova CO-48 CO2 Incubator, U.S. New BrunswickScientific CO., INC.), ultra-fine homogenizer (German Fluko company product), high speed low temperature centrifugal machine (Centrifuge5810R, Germany eppendorf company product),-80 ℃ of cryogenic refrigerator (Bio-Freezer, U.S. Forma Scientific company product), electronic balance (AE260-S, Switzerland Mettier company product), 24 well culture plates (Sigma, USA).
Chemical reagent: 199 (Sigma, USA), L-glutaminate (Fluka import packing), 30% hydrogen peroxide (H2O2) Luoyang City's chemical reagent factory product (040920).Penicillin injection (lot number: 0602010,060207 Huabei Pharmaceutic Co., Ltd's product), superoxide dismutase (SOD) testing cassete (lot number: 20070315), Coomassie brilliant blue protein determination kit (lot number: 20070626), (lot number: 20070626), the said determination test kit all builds up bio-engineering research institute available from Nanjing to the biuret method protein determination kit Huabei Pharmaceutic Co., Ltd's product), streptomycin injection (lot number:.
Experimental agents: L-Carnosine is provided by Shenyang Xing Qi Pharmaceutical Co; Than Nuo Kexin sodium eye drop (catalin, Co., Ltd of the long-range pharmacy in Wuhan group product, lot number: 061117).
Animal: 108 of healthy cleaning level SD rats, Zhengzhou University's Experimental Animal Center provides, and male and female are not limit, weight 160-200g.
Experimental technique
1. lens culture exsomatizes
Put to death the SD rat with head-breaking, get its eyeball, reject the tissues such as muscle, fascia, behind 500U/ml penicillin normal saline flushing, eyeball is dipped in the 500U/ml penicillin normal saline, after superclean bench changes 500U/ml penicillin normal saline and soaks, cuts off sclera by rear utmost point section, take out crystalline lens, crystalline lens is put into contained 5 * 10 4U/L penicillin and 5 * 10 4The u/L streptomycin without (1.5ml is inserted without phenol red 199 culture medium in the every hole of 24 well culture plates, the crystalline lens in every hole, totally 12 plates) in phenol red 199 culture medium (PH7.4), put in 37 ℃, 95% humidity, 5%CO2 incubator and cultivate.Behind the 17h, take out and observe under black background, 216 crystalline lens all transparents can be used for experiment.
2.H 2O 2The foundation of cataract model and grouping drug test
To be divided at random following 6 groups such as 216 transparent crystalline lenses of above-mentioned cultivation, 36 every group, each is organized crystalline lens and puts into respectively the sterile petri dish that contains following medicine and cultivate:
(1) matched group: serum-free is without 199 phenol red culture medium;
(2) model group: contain the serum-free of hydrogen peroxidase 10 .06mg/ml without 199 phenol red culture medium;
(3) 0.2% L-Carnosine groups: contain the serum-free of hydrogen peroxidase 10 .06mg/ml and 0.2% L-Carnosine (clinical medicine dose 1/5) without 199 phenol red culture medium;
(4) 1% L-Carnosine groups: contain the serum-free of hydrogen peroxidase 10 .06mg/ml and 1% L-Carnosine (clinical medicine dose) without 199 phenol red culture medium;
(5) 5% L-Carnosine groups: the nothing that contains hydrogen peroxidase 10 .06mg/ml and 5% L-Carnosine (clinical medicine dose 5 times) is clear without 199 phenol red culture medium;
(6) catalin group: contain the serum-free of hydrogen peroxidase 10 .06mg/ml and 1.06mg/100ml catalin (clinical medicine dose 1/5) without 199 phenol red culture medium.
Each is organized culture medium and all adds 5 * 104u/L penicillin and 5 * 104u/L streptomycin and 1%L-glutamine.Put and continue in 37 ℃, 95% humidity, 5%CO2 incubator to cultivate.Each is organized every 48h and changes once corresponding culture fluid; Except matched group, respectively organize every 24h and replenish 3% hydrogen peroxide, 3 μ, 1/ hole.
Respectively at 2,4,6 days observation lenticular opacity situations after the medication, according to following standard scoring.
3. lenticular opacity situation standards of grading
Design one wide 0.2mm under white background, the black lines of the square crossing of spacing 9mm places tested crystalline lens on the black lines of " ten " word intersection, gives classification with the definition that can see through crystalline lens." I " svelteness as seen, crystalline lens is fully transparent." II " lines are fuzzy, but profile still can distinguish, are slight muddiness." III " outline is unclear, and only central part mays be seen indistinctly, and is the moderate muddiness." IV " lines lose fully, for crystalline lens fully muddy.At last with each group len's opacity classification row statistical analysis.
4. water-solubility protein mensuration, insoluble protein assay and SOD determination of activity
Respectively at after the medication 2,4,6 days, from each group, get 12 crystalline lenses, to weigh ,-60 ℃ of refrigerator freezings are preserved, and are used for water-solubility protein and insoluble protein content and SOD determination of activity.
Amount by 1: 19 (W/V) during mensuration adds cold saline, with homogenate under the ultra-fine homogenizer ice bath, makes 5% crystalline lens homogenate, 2500r/min, and centrifugal 10min gets supernatant and dilutes on request rear mensuration water-soluble protein content and SOD activity.
With the amount adding cold saline of every pipe crystalline lens homogenate precipitation by 1: 9 (W/V), insoluble protein is dissolved with 56.56% carbamide, measure insoluble protein content, SOD is active with the insoluble protein cubage.Strictly press the experimental procedure operation of testing cassete explanation in the experiment.
5. statistical procedures: use the SPSS10.0 statistical software, the In Grade data adopts the Wilcoxon rank test, uses the Excel statistical software that quantitative data is adopted the t check, take P＜0.05 as difference statistical significance is arranged.
Test 1 L-Carnosine for the impact of lenticular opacity
1, lenticular opacity situation during medication was respectively organized in 2 days afterwards
According to the previous experiments method, experimental rat lenticular opacity situation during the observation medication was respectively organized in 2 days afterwards specifically sees Table 1.
Table 1 medication was respectively organized experimental rat lenticular opacity situation in 2 days relatively
Figure 2007101616010A008001010
Each test group result's statistical significance compares:
Matched group and model group P＜0.0005
Matched group and 0.2% L-Carnosine group P＜0.0005
Matched group and 1% L-Carnosine group P＜0.0005
Matched group and 5% L-Carnosine group P＜0.0005
Matched group and catalin group P＜0.0005
Model group and 0.2% L-Carnosine group P＞0.05
Model group and 1% L-Carnosine group P＜0.0005
Model group and 5% L-Carnosine group P＜0.0005
Model group and catalin group P＞0.05
0.2% L-Carnosine group and 1% L-Carnosine group P＜0.0005
0.2% L-Carnosine group and 5% L-Carnosine group P＜0.0005
0.2% L-Carnosine group and catalin group P＞0.05
1% L-Carnosine group and 5% L-Carnosine group P＞0.05
1% L-Carnosine group and catalin group P＜0.0005
5% L-Carnosine group and catalin group P＜0.0005
Respectively organize experimental rat lenticular opacity degree in 2 days by above-mentioned medication and learn by statistics processing as can be known:
Model group, 0.2% L-Carnosine group, 1% L-Carnosine group, 5% L-Carnosine group and catalin group lenticular opacity degree are apparently higher than matched group (P＜0.0005);
1% L-Carnosine group, 5% L-Carnosine group lenticular opacity degree are lower than respectively model group (P＜0.0005), 0.2% L-Carnosine group (P＜0.0005) and catalin group (P＜0.0005);
5% L-Carnosine group lenticular opacity degree and 1% L-Carnosine group there was no significant difference (P＞0.05)); 0.2% L-Carnosine group and catalin group lenticular opacity degree and model group there was no significant difference (P＞0.05); 0.2% L-Carnosine group lenticular opacity degree and catalin group there was no significant difference (P＞0.05).
2, lenticular opacity situation during medication was respectively organized in 4 days afterwards
According to the previous experiments method, experimental rat lenticular opacity situation during the observation medication was respectively organized in 4 days afterwards specifically sees Table 2.
Table 2 medication was respectively organized experimental rat lenticular opacity situation in 4 days relatively
Figure 2007101616010A00800011
Each test group result's statistical significance compares:
Matched group and model group P＜0.0005
Matched group and 0.2% L-Carnosine group P＜0.0005
Matched group and 1% L-Carnosine group P＜0.0005
Matched group and 5% L-Carnosine group P＜0.0005
Matched group and catalin group P＜0.0005
Model group and 0.2% L-Carnosine group P＞0.05
Model group and 1% L-Carnosine group P＜0.0005
Model group and 5% L-Carnosine group P＜0.0005
Model group and catalin group P＞0.05
0.2% L-Carnosine group and 1% L-Carnosine group P＜0.0005
0.2% L-Carnosine group and 5% L-Carnosine group P＜0.0005
0.2% L-Carnosine group and catalin group P＞0.05
1% L-Carnosine group and 5% L-Carnosine group P＜0.05
1% L-Carnosine group and catalin group P＜0.0005
5% L-Carnosine group and catalin group P＜0.0005
Respectively organized afterwards experimental rat lenticular opacity degree statistical result in 4 days as can be known by above-mentioned medication:
Model group, 0.2% L-Carnosine group, 1% L-Carnosine group, 5% L-Carnosine group and catalin group lenticular opacity degree are apparently higher than matched group (P＜0.0005);
1% L-Carnosine group, 5% L-Carnosine group lenticular opacity degree are lower than respectively model group (P＜0.0005), 0.2% L-Carnosine group, (P＜0.0005) and catalin group (P＜0.0005);
5% L-Carnosine group lenticular opacity degree is lower than and 1% L-Carnosine group (P＜0.05);
0.2% L-Carnosine group and catalin group lenticular opacity degree and model group there was no significant difference (P＞0.05); 0.2% L-Carnosine group lenticular opacity degree and catalin group there was no significant difference (P＞0.05).
3, lenticular opacity situation during medication was respectively organized in 6 days afterwards
According to the previous experiments method, experimental rat lenticular opacity situation during the observation medication was respectively organized in 6 days afterwards specifically sees Table 3.
Table 3 medication was respectively organized experimental rat lenticular opacity situation in 6 days relatively
Figure 2007101616010A00800021
Each test group result's statistical significance relatively
Matched group and model group P＜0.0005
Matched group and 0.2% L-Carnosine group P＜0.0005
Matched group and 1% L-Carnosine group P＜0.0005
Matched group and 5% L-Carnosine group P＜0.0005
Matched group and catalin group P＜0.0005
Model group and 0.2% L-Carnosine group P＞0.05
Model group and 1% L-Carnosine group P＜0.05
Model group and 5% L-Carnosine group P＜0.05
Model group and catalin group P＞0.05
0.2% L-Carnosine group and 1% L-Carnosine group P＜0.05
0.2% L-Carnosine group and 5% L-Carnosine group P＜0.0005
0.2% L-Carnosine group and catalin group P＞0.05
1% L-Carnosine group and 5% L-Carnosine group P＞0.05
1% L-Carnosine group and catalin group P＜0.05
5% L-Carnosine group and catalin group P＜0.0005
The statistical result of respectively organizing experimental rat lenticular opacity degree in 6 days by above-mentioned medication is as can be known:
Model group, 0.2% L-Carnosine group, 1% L-Carnosine group, 5% L-Carnosine group and catalin group lenticular opacity degree are apparently higher than matched group (P＜0.0005);
1% L-Carnosine group, 5% L-Carnosine group lenticular opacity degree are lower than respectively model group (P＜0.05), 0.2% L-Carnosine group (P＜0.05, P＜0.0005) and catalin group (P＜0.05, P＜0.0005);
5% L-Carnosine group lenticular opacity degree and 1% L-Carnosine group there was no significant difference (P＞0.05);
0.2% L-Carnosine group and catalin group lenticular opacity degree and model group there was no significant difference (P＞0.05), 0.2% L-Carnosine group lenticular opacity degree and catalin group there was no significant difference (P＞0.05).
The above results shows: after the medication 2-6 days, each group of except matched group other is with the oxidative injury time lengthening, len's opacity increases the weight of gradually, and 1% L-Carnosine group and 5% L-Carnosine group lenticular opacity degree are starkly lower than other each experimental grouies, show that 1% L-Carnosine and 5% L-Carnosine all have the effect that delays development of cataracts.
Test 2 L-Carnosines for the impact of crystalline lens water-soluble protein content
1, the variation of crystalline lens water-soluble protein content during medication was respectively organized in 2 days afterwards
According to the previous experiments method, experimental rat crystalline lens water-soluble protein content situation of change during the observation medication was respectively organized in 2 days afterwards specifically sees Table 4.
The comparison (x+s) of crystalline lens water-soluble protein content was respectively organized in table 4 medication in 2 days
Figure 2007101616010A00800031
Each test group result's statistical significance compares:
Matched group and model group P＜0.005
Matched group and 0.2% L-Carnosine group P＜0.005
Matched group and 1% L-Carnosine group P＜0.005
Matched group and 5% L-Carnosine group P＜0.005
Matched group and catalin group P＜0.005
Model group and 0.2% L-Carnosine group P＞0.05
Model group and 1% L-Carnosine group P＜0.0005
Model group and 5% L-Carnosine group P＜0.0005
Model group and catalin group P＞0.05
0.2% L-Carnosine group and 1% L-Carnosine group P＜0.05
0.2% L-Carnosine group and 5% L-Carnosine group P＜0.01
0.2% L-Carnosine group and catalin group P＞0.05
1% L-Carnosine group and 5% L-Carnosine group P＞0.05
1% L-Carnosine group and catalin group P＜0.001
5% L-Carnosine group and catalin group P＜0.0005
Medication was respectively organized experimental rat crystalline lens water-soluble protein content in 2 days and is learned by statistics processes and displays, model group, 0.2% L-Carnosine group, 1% L-Carnosine group, 5% L-Carnosine group and catalin group crystalline lens water-soluble protein content are starkly lower than matched group (P＜0.0005), 1% L-Carnosine group and 5% L-Carnosine group crystalline lens water-soluble protein content are higher than respectively model group (P＜0.0005), 0.2% L-Carnosine group (P＜0.05, P＜0.01) and catalin group (P＜0.001, P＜0.0005), 5% L-Carnosine group crystalline lens water-soluble protein content and 1% L-Carnosine group there was no significant difference (P＞0.05), 0.2% L-Carnosine group and catalin group crystalline lens water-soluble protein content and model group there was no significant difference (P＞0.05), 0.2% L-Carnosine group crystalline lens water-soluble protein content and catalin group there was no significant difference (P＞0.05).
2, the variation of crystalline lens water-soluble protein content during medication was respectively organized in 4 days afterwards
According to the previous experiments method, experimental rat crystalline lens water-soluble protein content situation of change during the observation medication was respectively organized in 4 days afterwards specifically sees Table 5.
The comparison (x+s) of crystalline lens water-soluble protein content was respectively organized in table 5 medication in 4 days
Figure 2007101616010A00800041
Each test group result's statistical significance compares:
Matched group and model group P＜0.005
Matched group and 0.2% L-Carnosine group P＜0.005
Matched group and 1% L-Carnosine group P＜0.005
Matched group and 5% L-Carnosine group P＜0.005
Matched group and catalin group P＜0.005
Model group and 0.2% L-Carnosine group P＞0.05
Model group and 1% L-Carnosine group P＜0.05
Model group and 5% L-Carnosine group P＜0.005
Model group and catalin group P＞0.05
0.2% L-Carnosine group and 1% L-Carnosine group P＜0.05
0.2% L-Carnosine group and 5% L-Carnosine group P＜0.005
0.2% L-Carnosine group and catalin group P＞0.05
1% L-Carnosine group and 5% L-Carnosine group P＞0.05
1% L-Carnosine group and catalin group P＜0.05
5% L-Carnosine group and catalin group P＜0.005
Medication was respectively organized experimental rat crystalline lens water-soluble protein content in 4 days and is learned by statistics processes and displays, model group, 0.2% L-Carnosine group, 1% L-Carnosine group, 5% L-Carnosine group and catalin group crystalline lens water-soluble protein content are starkly lower than matched group (P＜0.0005), 1% L-Carnosine group and 5% L-Carnosine group crystalline lens water-soluble protein content are higher than respectively model group (P＜0.05, P＜0.005), 0.2% L-Carnosine group (P＜0.05, P＜0.005) and catalin group (P＜0.05, P＜0.005), 5% L-Carnosine group crystalline lens water-soluble protein content and 1% L-Carnosine group there was no significant difference (P＞0.05), 0.2% L-Carnosine group and catalin group crystalline lens water-soluble protein content and model group there was no significant difference (P＞0.05), 0.2% L-Carnosine group crystalline lens water-soluble protein content and catalin group there was no significant difference (P＞0.05).
3, the variation of crystalline lens water-soluble protein content during medication was respectively organized in 6 days afterwards
According to the previous experiments method, experimental rat crystalline lens water-soluble protein content situation of change during the observation medication was respectively organized in 6 days afterwards specifically sees Table 6.
The comparison (x+s) of crystalline lens water-soluble protein content was respectively organized in table 6 medication in 6 days
Figure 2007101616010A00800051
Each test group result's statistical significance compares:
Matched group and model group P＜0.0005
Matched group and 0.2% L-Carnosine group P＜0.0005
Matched group and 1% L-Carnosine group P＜0.0005
Matched group and 5% L-Carnosine group P＜0.0005
Matched group and catalin group P＜0.0005
Model group and 0.2% L-Carnosine group P＞0.05
Model group and 1% L-Carnosine group P＜0.05
Model group and 5% L-Carnosine group P＜0.0005
Model group and catalin group P＞0.05
0.2% L-Carnosine group and 1% L-Carnosine group P＜0.01
0.2% L-Carnosine group and 5% L-Carnosine group P＜0.0005
0.2% L-Carnosine group and catalin group P＞0.05
1% L-Carnosine group and 5% L-Carnosine group P＜0.0005
1% L-Carnosine group and catalin group P＜0.005
5% L-Carnosine group and catalin group P＜0.0005
Medication was respectively organized experimental rat crystalline lens water-soluble protein content in 6 days and is learned by statistics processes and displays, and model group, 0.2% L-Carnosine group, 1% L-Carnosine group, 5% L-Carnosine group and catalin group crystalline lens water-soluble protein content are starkly lower than matched group (P＜0.0005);
1% L-Carnosine group and 5% L-Carnosine group crystalline lens water-soluble protein content are higher than respectively model group (P＜0.05, P＜0.0005), 0.2% L-Carnosine group (P＜0.01, P＜0.0005) and catalin group (P＜0.005, P＜0.0005);
5% L-Carnosine group crystalline lens water-soluble protein content is higher than 1% L-Carnosine group (P＜0.0005);
0.2% L-Carnosine group and catalin group crystalline lens water-soluble protein content and model group there was no significant difference (P＞0.05), 0.2% L-Carnosine group crystalline lens water-soluble protein content and catalin group there was no significant difference (P＞0.05).
The above results shows: after the medication 2-6 days, each organized other except matched group with the oxidative injury time lengthening, and the crystalline lens water-soluble protein content reduces gradually.
Lenticular water-solubility protein is lenticular structural protein, and is close with the maintain the relationship of the lenticular transparency.In the Cataractogenesis process, water-soluble protein content reduces.And 1% L-Carnosine group and 5% L-Carnosine group crystalline lens water-soluble protein content show that apparently higher than other each experimental grouies 1% L-Carnosine and 5% L-Carnosine all have the effect that delays development of cataracts.
Test 3 L-Carnosines to the impact of SOD activity in the crystalline lens
1, the variation of SOD activity in the crystalline lens was respectively organized in medication in 2 days afterwards
According to the previous experiments method, the situation of change of SOD activity in the experimental rat crystalline lens specifically saw Table 7 during the observation medication was respectively organized in 2 days afterwards.
The comparison (x+s) of crystalline lens SOD activity was respectively organized in table 7 medication in 2 days
Figure 2007101616010A00800061
Each test group result's statistical significance compares:
Matched group and model group P＜0.0005
Matched group and 0.2% L-Carnosine group P＜0.0005
Matched group and 1% L-Carnosine group P＜0.0005
Matched group and 5% L-Carnosine group P＜0.0005
Matched group and catalin group P＜0.0005
Model group and 0.2% L-Carnosine group P＞0.05
Model group and 1% L-Carnosine group P＜0.001
Model group and 5% L-Carnosine group P＜0.005
Model group and catalin group P＞0.05
0.2% L-Carnosine group and 1% L-Carnosine group P＜0.005
0.2% L-Carnosine group and 5% L-Carnosine group P＜0.05
0.2% L-Carnosine group and catalin group P＞0.05
1% L-Carnosine group and 5% L-Carnosine group P＞0.05
1% L-Carnosine group and catalin group P＜0.005
5% L-Carnosine group and catalin group P＜0.001
Medication was respectively organized experimental rat crystalline lens SOD activity in 2 days and is learned by statistics processes and displays:
Model group, 0.2% L-Carnosine group, 1% L-Carnosine group, 5% L-Carnosine group and catalin group crystal SOD activity are starkly lower than matched group (P＜0.0005);
1% L-Carnosine group and 5% L-Carnosine group crystal SOD activity be respectively apparently higher than model group (P＜0.001, P＜0.005), 0.2% L-Carnosine group (P＜0.005, P＜0.05) and catalin group (P＜0.005, P＜0.001);
0.2% L-Carnosine group, catalin crystal SOD activity and model group there was no significant difference (P＞0.05); Active and the catalin group there was no significant difference (P＞0.05) of 0.2% L-Carnosine group crystal SOD; Active and the 5% L-Carnosine group there was no significant difference (P＞0.05) of 1% L-Carnosine group crystal SOD.
2, the variation of SOD activity in the crystalline lens was respectively organized in medication in 4 days afterwards
According to the previous experiments method, the situation of change of SOD activity in the experimental rat crystalline lens specifically saw Table 8 during the observation medication was respectively organized in 4 days afterwards.
The comparison (x+s) of crystalline lens SOD activity was respectively organized in table 8 medication in 4 days
Figure 2007101616010A00800071
Each test group result's statistical significance compares:
Matched group and model group P＜0.0005
Matched group and 0.2% L-Carnosine group P＜0.0005
Matched group and 1% L-Carnosine group P＜0.0005
Matched group and 5% L-Carnosine group P＜0.0005
Matched group and catalin group P＜0.0005
Model group and 0.2% L-Carnosine group P＜0.0005
Model group and 1% L-Carnosine group P＜0.0005
Model group and 5% L-Carnosine group P＜0.0005
Model group and catalin group P＜0.0005
0.2% L-Carnosine group and 1% L-Carnosine group P＜0.05
0.2% L-Carnosine group and 5% L-Carnosine group P＜0.01
0.2% L-Carnosine group and catalin group P＞0.05
1% L-Carnosine group and 5% L-Carnosine group P＞0.05
1% L-Carnosine group and catalin group P＜0.01
5% L-Carnosine group and catalin group P＜0.05
Medication was respectively organized experimental rat crystalline lens SOD activity in 4 days and is learned by statistics processes and displays:
Model group, 0.2% L-Carnosine group, 1% L-Carnosine group, 5% L-Carnosine group and catalin group crystal SOD activity are starkly lower than matched group (P＜0.0005);
0.2% L-Carnosine group, 1% L-Carnosine group, 5% L-Carnosine group and catalin group crystal SOD activity are apparently higher than model group (P＜0.0005);
1% L-Carnosine group and 5% L-Carnosine group crystal SOD activity are higher than respectively catalin (P＜0.01, P＜0.05) and 0.2% L-Carnosine group (P＜0.05, P＜0.01);
Active and the 5% L-Carnosine group there was no significant difference (P＞0.05) of 1% L-Carnosine group crystal SOD; Active and the catalin group there was no significant difference (P＞0.05) of 0.2% L-Carnosine group crystal SOD.
3, the variation of SOD activity in the crystalline lens was respectively organized in medication in 6 days afterwards
According to the previous experiments method, the situation of change of SOD activity in the experimental rat crystalline lens specifically saw Table 9 during the observation medication was respectively organized in 6 days afterwards.
The comparison (x+s) of crystalline lens SOD activity was respectively organized in table 9 medication in 6 days
Figure 2007101616010A00800081
Each test group result's statistical significance compares:
Matched group and model group P＜0.0005
Matched group and 0.2% L-Carnosine group P＜0.0005
Matched group and 1% L-Carnosine group P＜0.0005
Matched group and 5% L-Carnosine group P＜0.0005
Matched group and catalin group P＜0.0005
Model group and 0.2% L-Carnosine group P＜0.0005
Model group and 1% L-Carnosine group P＜0.0005
Model group and 5% L-Carnosine group P＜0.0005
Model group and catalin group P＜0.005
0.2% L-Carnosine group and 1% L-Carnosine group P＜0.05
0.2% L-Carnosine group and 5% L-Carnosine group P＜0.005
0.2% L-Carnosine group and catalin group P＞0.05
1% L-Carnosine group and 5% L-Carnosine group P＞0.05
1% L-Carnosine group and catalin group P＜0.05
5% L-Carnosine group and catalin group P＜0.05
Medication was respectively organized experimental rat crystalline lens SOD activity in 6 days and is learned by statistics processes and displays:
Model group, 0.2% L-Carnosine group, 1% L-Carnosine group, 5% L-Carnosine group and catalin group crystal SOD activity are starkly lower than matched group (P＜0.0005);
0.2% L-Carnosine group, 1% L-Carnosine group, 5% L-Carnosine group and catalin group crystal SOD activity are apparently higher than model group (P＜0.0005);
1% L-Carnosine group and 5% L-Carnosine group group crystal SOD activity are higher than respectively catalin (P＜0.05) and 0.2% L-Carnosine group (P＜0.05, P＜0.005);
Active and the 5% L-Carnosine group there was no significant difference (P＞0.05) of 1% L-Carnosine group crystal SOD; Active and the catalin group there was no significant difference (P＞0.05) of 0.2% L-Carnosine group crystal SOD.
The above results shows: after the medication 2-6 days, each group of other except matched group was with the oxidative injury time lengthening, and crystalline lens SOD activity reduces gradually, points out lipid peroxide and radical pair crystalline lens activities of antioxidant enzymes inhibited.And 1% L-Carnosine group and 5% L-Carnosine group crystalline lens SOD activity show that apparently higher than other each experimental grouies external additional 1% L-Carnosine or 5% L-Carnosine can obviously improve the activity of crystalline lens antioxidase, delay the progress of rat cataract.
By above-mentioned test 1-3 as can be known:
1) after the medication 2-6 day, 1% L-Carnosine group and 5% L-Carnosine group lenticular opacity degree are starkly lower than other each experimental grouies (P＜0.05-0.0005), (P＜0.05-0.0005), crystalline lens SOD activity is apparently higher than other each experimental grouies (P＜0.05-0.0005) apparently higher than other each experimental grouies for the crystalline lens water-soluble protein content.Point out 1% L-Carnosine and 5% L-Carnosine can improve the activity of crystalline lens antioxidase, alleviate lenticular oxidative damage, the effect that delays development of cataracts is arranged.
2) after the medication 2-6 days, 1% L-Carnosine group lenticular opacity degree, crystalline lens water-soluble protein content and crystalline lens SOD active with 5% L-Carnosine group there was no significant difference (P＞0.05).
3) after the medication 2-6 days, 0.2% L-Carnosine (clinical medicine dose 1/5) group and catalin (clinical medicine dose 1/5) group lenticular opacity degree, crystalline lens water-soluble protein content and crystalline lens SOD is active and model group there was no significant difference (P＞0.05).
4) after the medication 2-6 days, 0.2% L-Carnosine (clinical medicine dose 1/5) group lenticular opacity degree, crystalline lens water-soluble protein content and crystalline lens SOD active with catalin (clinical medicine dose 1/5) group there was no significant difference (P＞0.05).
5) activity of L-Carnosine increases with the raising of concentration.
The pharmaceutical composition that embodiment 2 contains L-Carnosine delays the activity of development of cataracts
Experiment material, experimental model animal and stripped lens culture and H 2O 2The method of the foundation of cataract model is with embodiment 1.Wherein concrete Experimental agents comprises:
1) 5% L-Carnosine
Carnosine 50g, surplus is pure water, is mixed with the 1000g eye drop.
2) compositions A
L-Carnosine 50g, ethyl hydroxybenzoate 0.3g, hyaluronic acid sodium 0.5g controls pH value with boric acid-Borax between 6.8 to 8.0 as buffer salt, regulates osmotic pressure to 260-340mosm/L with an amount of sodium chloride, and surplus is pure water, is mixed with the 1000g eye drop.
To be divided at random following 5 groups such as 180 transparent crystalline lenses of aforementioned cultivation, 36 every group, each is organized crystalline lens and puts into respectively the sterile petri dish that contains following medicine and cultivate:
(1) matched group: serum-free is without 199 phenol red culture medium;
(2) model group: contain the serum-free of hydrogen peroxidase 10 .06mg/ml without 199 phenol red culture medium;
(3) 5% L-Carnosine groups: contain the serum-free of hydrogen peroxidase 10 .06mg/ml and 1% L-Carnosine (clinical medicine dose) without 199 phenol red culture medium;
(4) compositions A group: the nothing that contains hydrogen peroxidase 10 .06mg/ml and compositions A is clear without 199 phenol red culture medium;
(5) catalin group: contain the serum-free of hydrogen peroxidase 10 .06mg/ml and 1.06mg/100ml catalin (clinical medicine dose 1/5) without 199 phenol red culture medium.
Each is organized culture medium and all adds 5 * 104u/L penicillin and 5 * 104u/L streptomycin and 1%L-glutamine.Put and continue in 37 ℃, 95% humidity, 5%CO2 incubator to cultivate.Each is organized every 48h and changes once corresponding culture fluid; Except matched group, respectively organize every 24h and replenish 3% hydrogen peroxide, 3 μ l/ holes.
Respectively at 2,4,6 days observation lenticular opacity situations after the medication, according to following standard scoring.
3. lenticular opacity situation standards of grading
Referring to embodiment 1.
4. water-solubility protein mensuration, insoluble protein assay and SOD determination of activity
Referring to embodiment 1.
5. statistical procedures: use the SPSS10.0 statistical software, the In Grade data adopts the Wilcoxon rank test, uses the Excel statistical software that quantitative data is adopted the t check, take P＜0.05 as difference statistical significance is arranged.
Test 1. compositions A for the impact of lenticular opacity
1, lenticular opacity situation during medication was respectively organized in 2 days afterwards
According to the previous experiments method, experimental rat lenticular opacity situation during the observation medication was respectively organized in 2 days afterwards specifically sees Table 10.
Table 10 medication was respectively organized experimental rat lenticular opacity situation in 2 days relatively
Figure 2007101616010A00800091
Each test group result's statistical significance compares:
Matched group and model group P＜0.0005
Matched group and 5% L-Carnosine group P＜0.0005
Matched group and compositions A group P＜0.0005
Matched group and catalin group P＜0.0005
Model group and 5% L-Carnosine group P＜0.0005
Model group and compositions A group P＜0.0005
Model group and catalin group P＞0.05
5% L-Carnosine group and compositions A group P＞0.05
5% L-Carnosine group and catalin group P＜0.0005
Compositions A and catalin group P＜0.0005
Respectively organize experimental rat lenticular opacity degree in 2 days by above-mentioned medication and learn by statistics processing as can be known:
Model group, 5% L-Carnosine group, compositions A group and catalin group lenticular opacity degree are apparently higher than matched group (P＜0.0005);
5% L-Carnosine group, compositions A group lenticular opacity degree are lower than respectively model group (P＜0.0005); Compositions A group lenticular opacity degree and 5% L-Carnosine group there was no significant difference (P＞0.05).
2, lenticular opacity situation during medication was respectively organized in 4 days afterwards
According to the previous experiments method, experimental rat lenticular opacity situation during the observation medication was respectively organized in 4 days afterwards specifically sees Table 11.
Table 11 medication was respectively organized experimental rat lenticular opacity situation in 4 days relatively
Figure 2007101616010A00800101
Each test group result's statistical significance compares:
Matched group and model group P＜0.0005
Matched group and 5% L-Carnosine group P＜0.0005
Matched group and compositions A group P＜0.0005
Matched group and catalin group P＜0.0005
Model group and 5% L-Carnosine group P＜0.0005
Model group and compositions A group P＜0.0005
Model group and catalin group P＞0.05
5% L-Carnosine group and compositions A group P＜0.05
5% L-Carnosine group and catalin group P＜0.0005
Compositions A group and catalin group P＜0.0005
The statistical result of respectively organizing afterwards experimental rat lenticular opacity degree in 4 days by above-mentioned medication is as can be known:
Model group, 5% L-Carnosine group, compositions A group and catalin group lenticular opacity degree are apparently higher than matched group (P＜0.0005);
5% L-Carnosine group, compositions A group lenticular opacity degree are lower than respectively model group (P＜0.0005); Compositions A group lenticular opacity degree is lower than and 5% L-Carnosine group (P＜0.05).
3, lenticular opacity situation during medication was respectively organized in 6 days afterwards
According to the previous experiments method, experimental rat lenticular opacity situation during the observation medication was respectively organized in 6 days afterwards specifically sees Table 12.
Table 12 medication was respectively organized experimental rat lenticular opacity situation in 6 days relatively
Figure 2007101616010A00800111
Each test group result's statistical significance relatively
Matched group and model group P＜0.0005
Matched group and 5% L-Carnosine group P＜0.0005
Matched group and compositions A group P＜0.0005
Matched group and catalin group P＜0.0005
Model group and 5% L-Carnosine group P＜0.05
Model group and compositions A group P＜0.05
Model group and catalin group P＞0.05
5% L-Carnosine group and compositions A group P＞0.05
5% L-Carnosine group and catalin group P＜0.05
Compositions A group and catalin group P＜0.0005
The statistical result of respectively organizing experimental rat lenticular opacity degree in 6 days by above-mentioned medication is as can be known:
Model group, 5% L-Carnosine group, compositions A group and catalin group lenticular opacity degree are apparently higher than matched group (P＜0.0005);
5% L-Carnosine group, compositions A group lenticular opacity degree are lower than respectively model group (P＜0.05); Compositions A group lenticular opacity degree and 5% L-Carnosine group there was no significant difference (P＞0.05).
The above results shows: after the medication 2-6 days, each group of except matched group other is with the oxidative injury time lengthening, len's opacity increases the weight of gradually, and 5% L-Carnosine group and compositions A group lenticular opacity degree are starkly lower than other each experimental grouies, show that 5% L-Carnosine and compositions A have the effect that delays development of cataracts.
Test 2 compositions A for the impact of crystalline lens water-soluble protein content
1, the variation of crystalline lens water-soluble protein content during medication was respectively organized in 2 days afterwards
According to the previous experiments method, experimental rat crystalline lens water-soluble protein content situation of change during the observation medication was respectively organized in 2 days afterwards specifically sees Table 13.
The comparison (x+s) of crystalline lens water-soluble protein content was respectively organized in table 13 medication in 2 days
Each test group result's statistical significance compares:
Matched group and model group P＜0.005
Matched group and 5% L-Carnosine group P＜0.005
Matched group and compositions A group P＜0.005
Matched group and catalin group P＜0.005
Model group and 5% L-Carnosine group P＜0.0005
Model group and compositions A group P＜0.0005
Model group and catalin group P＞0.05
5% L-Carnosine group and compositions A group P＞0.05
5% L-Carnosine group and catalin group P＜0.001
Compositions A group and catalin group P＜0.0005
Medication was respectively organized experimental rat crystalline lens water-soluble protein content in 2 days and is learned by statistics processes and displays, model group, 5% L-Carnosine group, compositions A group and catalin group crystalline lens water-soluble protein content are starkly lower than matched group (P＜0.0005), 5% L-Carnosine group and compositions A group crystalline lens water-soluble protein content are higher than respectively model group (P＜0.0005), compositions A group crystalline lens water-soluble protein content and 5% L-Carnosine group there was no significant difference (P＞0.05).
2, the variation of crystalline lens water-soluble protein content during medication was respectively organized in 4 days afterwards
According to the previous experiments method, experimental rat crystalline lens water-soluble protein content situation of change during the observation medication was respectively organized in 4 days afterwards specifically sees Table 14.
The comparison (x+s) of crystalline lens water-soluble protein content was respectively organized in table 14 medication in 4 days
Figure 2007101616010A00800131
Each test group result's statistical significance compares:
Matched group and model group P＜0.005
Matched group and 5% L-Carnosine group P＜0.005
Matched group and compositions A group P＜0.005
Matched group and catalin group P＜0.005
Model group and 5% L-Carnosine group P＜0.05
Model group and compositions A group P＜0.005
Model group and catalin group P＞0.05
5% L-Carnosine group and compositions A group P＞0.05
5% L-Carnosine group and catalin group P＜0.05
Compositions A group and catalin group P＜0.005
Medication was respectively organized experimental rat crystalline lens water-soluble protein content in 4 days and is learned by statistics processes and displays, model group, 5% L-Carnosine group, compositions A group and catalin group crystalline lens water-soluble protein content are starkly lower than matched group (P＜0.0005), 5% L-Carnosine group and compositions A group crystalline lens water-soluble protein content are higher than respectively model group (P＜0.05, P＜0.005) and catalin group (P＜0.05, P＜0.005), compositions A group crystalline lens water-soluble protein content and 5% L-Carnosine group there was no significant difference (P＞0.05).
3, the variation of crystalline lens water-soluble protein content during medication was respectively organized in 6 days afterwards
According to the previous experiments method, experimental rat crystalline lens water-soluble protein content situation of change during the observation medication was respectively organized in 6 days afterwards specifically sees Table 15.
The comparison (x+s) of crystalline lens water-soluble protein content was respectively organized in table 15 medication in 6 days
Figure 2007101616010A00800141
Each test group result's statistical significance compares:
Matched group and model group P＜0.0005
Matched group and 5% L-Carnosine group P＜0.0005
Matched group and compositions A group P＜0.0005
Matched group and catalin group P＜0.0005
Model group and 5% L-Carnosine group P＜0.05
Model group and compositions A group P＜0.0005
Model group and catalin group P＞0.05
5% L-Carnosine group and compositions A group P＜0.0005
5% L-Carnosine group and catalin group P＜0.005
Compositions A group and catalin group P＜0.0005
Medication was respectively organized experimental rat crystalline lens water-soluble protein content in 6 days and is learned by statistics processes and displays, and model group, 5% L-Carnosine group, compositions A group and catalin group crystalline lens water-soluble protein content are starkly lower than matched group (P＜0.0005);
5% L-Carnosine group and compositions A group crystalline lens water-soluble protein content are higher than respectively model group (P＜0.05, P＜0.0005) and catalin group (P＜0.005, P＜0.0005);
Compositions A group crystalline lens water-soluble protein content is higher than 5% L-Carnosine (P＜0.0005).
The above results shows: after the medication 2-6 days, each organized other except matched group with the oxidative injury time lengthening, and the crystalline lens water-soluble protein content reduces gradually.
Lenticular water-solubility protein is lenticular structural protein, and is close with the maintain the relationship of the lenticular transparency.In the Cataractogenesis process, water-soluble protein content reduces.And 5% L-Carnosine group and compositions A group crystalline lens water-soluble protein content show that apparently higher than other each experimental grouies 5% L-Carnosine and compositions A have the effect that delays development of cataracts.
Test 3 compositions A to the impact of SOD activity in the crystalline lens
1, the variation of SOD activity in the crystalline lens was respectively organized in medication in 2 days afterwards
According to the previous experiments method, the situation of change of SOD activity in the experimental rat crystalline lens specifically saw Table 16 during the observation medication was respectively organized in 2 days afterwards.
The comparison (x+s) of crystalline lens SOD activity was respectively organized in table 16 medication in 2 days
Figure 2007101616010A00800151
Each test group result's statistical significance compares:
Matched group and model group P＜0.0005
Matched group and 5% L-Carnosine group P＜0.0005
Matched group and compositions A group P＜0.0005
Matched group and catalin group P＜0.0005
Model group and 5% L-Carnosine group P＜0.001
Model group and compositions A group P＜0.005
Model group and catalin group P＞0.05
5% L-Carnosine group and compositions A group P＞0.05
5% L-Carnosine group and catalin group P＜0.005
Compositions A group and catalin group P＜0.001
Medication was respectively organized experimental rat crystalline lens SOD activity in 2 days and is learned by statistics processes and displays:
Model group, 5% L-Carnosine group, compositions A group and catalin group crystal SOD activity are starkly lower than matched group (P＜0.0005);
5% L-Carnosine group and compositions A group crystal SOD activity are respectively apparently higher than model group (P＜0.001, P＜0.005) and catalin group (P＜0.005, P＜0.001); 5% L-Carnosine group crystal SOD is active to organize there was no significant difference (P＞0.05) with compositions A.
2, the variation of SOD activity in the crystalline lens was respectively organized in medication in 4 days afterwards
According to the previous experiments method, the situation of change of SOD activity in the experimental rat crystalline lens specifically saw Table 17 during the observation medication was respectively organized in 4 days afterwards.
The comparison (x+s) of crystalline lens SOD activity was respectively organized in table 17 medication in 4 days
Figure 2007101616010A00800161
Each test group result's statistical significance compares:
Matched group and model group P＜0.0005
Matched group and 5% L-Carnosine group P＜0.0005
Matched group and compositions A group P＜0.0005
Matched group and catalin group P＜0.0005
Model group and 5% L-Carnosine group P＜0.0005
Model group and compositions A group P＜0.0005
Model group and catalin group P＜0.0005
5% L-Carnosine group and compositions A group P＞0.05
5% L-Carnosine group and catalin group P＜0.01
Compositions A group and catalin group P＜0.05
Medication was respectively organized experimental rat crystalline lens SOD activity in 4 days and is learned by statistics processes and displays:
Model group, 5% L-Carnosine group, compositions A group and catalin group crystal SOD activity are starkly lower than matched group (P＜0.0005); 5% L-Carnosine group, compositions A group and catalin group crystal SOD activity are apparently higher than model group (P＜0.0005);
5% L-Carnosine group and compositions A group crystal SOD activity are higher than respectively catalin (P＜0.01, P＜0.05); 5% L-Carnosine group crystal SOD is active to organize there was no significant difference (P＞0.05) with compositions A.
3, the variation of SOD activity in the crystalline lens was respectively organized in medication in 6 days afterwards
According to the previous experiments method, the situation of change of SOD activity in the experimental rat crystalline lens specifically saw Table 18 during the observation medication was respectively organized in 6 days afterwards.
The comparison (x+s) of crystalline lens SOD activity was respectively organized in table 18 medication in 6 days
Figure 2007101616010A00800171
Each test group result's statistical significance compares:
Matched group and model group P＜0.0005
Matched group and 5% L-Carnosine group P＜0.0005
Matched group and compositions A group P＜0.0005
Matched group and catalin group P＜0.0005
Model group and 5% L-Carnosine group P＜0.0005
Model group and compositions A group P＜0.0005
Model group and catalin group P＜0.005
5% L-Carnosine group and compositions A group P＞0.05
5% L-Carnosine group and catalin group P＜0.05
Compositions A group and catalin group P＜0.05
Medication was respectively organized experimental rat crystalline lens SOD activity in 6 days and is learned by statistics processes and displays:
Model group, 5% L-Carnosine group, compositions A group and catalin group crystal SOD activity are starkly lower than matched group (P＜0.0005);
5% L-Carnosine group, compositions A group and catalin group crystal SOD activity are apparently higher than model group (P＜0.0005); 5% L-Carnosine group and compositions A group crystal SOD activity are higher than respectively catalin (P＜0.05); 5% L-Carnosine group crystal SOD is active to organize there was no significant difference (P＞0.05) with compositions A.
The above results shows: after the medication 2-6 days, each group of other except matched group was with the oxidative injury time lengthening, and crystalline lens SOD activity reduces gradually, points out lipid peroxide and radical pair crystalline lens activities of antioxidant enzymes inhibited.And 5% L-Carnosine group and compositions A group crystalline lens SOD activity show that apparently higher than other each experimental grouies external additional 5% L-Carnosine or compositions A can obviously improve the activity of crystalline lens antioxidase, delay the progress of rat cataract.
By above-mentioned test 1-3 as can be known:
1) after the medication 2-6 day, 5% L-Carnosine group and compositions A group lenticular opacity degree is starkly lower than other each experimental grouies (P＜0.05-0.0005), (P＜0.05-0.0005), crystalline lens SOD activity is apparently higher than other each experimental grouies (P＜0.05-0.0005) apparently higher than other each experimental grouies for the crystalline lens water-soluble protein content.Point out 5% L-Carnosine and compositions A can improve the activity of crystalline lens antioxidase, alleviate lenticular oxidative damage, the effect that delays development of cataracts is arranged.
2) after the medication 2-6 days, 5% L-Carnosine group lenticular opacity degree, crystalline lens water-soluble protein content and crystalline lens SOD active with compositions A group there was no significant difference (P＞0.05).
The eye-gel preparation that embodiment 3 contains L-Carnosine delays the activity of development of cataracts
Experiment material, experimental model animal and stripped lens culture and H 2O 2The method of the foundation of cataract model is with embodiment 1.Wherein concrete Experimental agents comprises:
5% L-Carnosine
L-Carnosine 50g, surplus is pure water, is mixed with the 1000g eye drop.
Eye-gel preparation is hereinafter referred to as compositions B
L-Carnosine 50g, sodium carboxymethyl cellulose 40g, ethyl hydroxybenzoate 0.3g, hyaluronic acid sodium 0.1g controls pH value with boric acid-Borax between 6.8 to 8.0 as buffer salt, regulates osmotic pressure to 260-340mosm/L with an amount of propylene glycol, surplus is pure water, is prepared into the 1000g gel for eye use.
To be divided at random following 5 groups such as 180 transparent crystalline lenses of aforementioned cultivation, 36 every group, each is organized crystalline lens and puts into respectively the sterile petri dish that contains following medicine and cultivate:
(1) matched group: serum-free is without 199 phenol red culture medium;
(2) model group: contain the serum-free of hydrogen peroxidase 10 .06mg/ml without 199 phenol red culture medium;
(3) 5% L-Carnosine groups: contain the serum-free of hydrogen peroxidase 10 .06mg/ml and 1% L-Carnosine (clinical medicine dose) without 199 phenol red culture medium;
(4) compositions B group: the nothing that contains hydrogen peroxidase 10 .06mg/ml and compositions A is clear without 199 phenol red culture medium;
(5) catalin group: contain the serum-free of hydrogen peroxidase 10 .06mg/ml and 1.06mg/100ml catalin (clinical medicine dose 1/5) without 199 phenol red culture medium.
Each is organized culture medium and all adds 5 * 104u/L penicillin and 5 * 104u/L streptomycin and 1%L-glutamine.Put and continue in 37 ℃, 95% humidity, 5%CO2 incubator to cultivate.Each is organized every 48h and changes once corresponding culture fluid; Except matched group, respectively organize every 24h and replenish 3% hydrogen peroxide, 3 μ l/ holes.
Respectively at 2,4,6 days observation lenticular opacity situations after the medication, according to following standard scoring.
3. lenticular opacity situation standards of grading
Referring to embodiment 1.
4. water-solubility protein mensuration, insoluble protein assay and SOD determination of activity
Referring to embodiment 1.
5. statistical procedures: use the SPSS10.0 statistical software, the In Grade data adopts the Wilcoxon rank test, uses the Excel statistical software that quantitative data is adopted the t check, take P＜0.05 as difference statistical significance is arranged.
Test 1. compositions B for the impact of lenticular opacity
1, lenticular opacity situation during medication was respectively organized in 2 days afterwards
According to the previous experiments method, experimental rat lenticular opacity situation during the observation medication was respectively organized in 2 days afterwards specifically sees Table 19.
Table 19 medication was respectively organized experimental rat lenticular opacity situation in 2 days relatively
Figure 2007101616010A00800181
Each test group result's statistical significance compares:
Matched group and model group P＜0.0005
Matched group and 5% L-Carnosine group P＜0.0005
Matched group and compositions B group P＜0.0005
Matched group and catalin group P＜0.0005
Model group and 5% L-Carnosine group P＜0.0005
Model group and compositions B group P＜0.0005
Model group and catalin group P＞0.05
5% L-Carnosine group and compositions B group P＞0.05
5% L-Carnosine group and catalin group P＜0.0005
Compositions B and catalin group P＜0.0005
Respectively organize experimental rat lenticular opacity degree in 2 days by above-mentioned medication and learn by statistics processing as can be known:
Model group, 5% L-Carnosine group, compositions B group and catalin group lenticular opacity degree are apparently higher than matched group (P＜0.0005);
5% L-Carnosine group, compositions B group lenticular opacity degree are lower than respectively model group (P＜0.0005); Compositions B group lenticular opacity degree and 5% L-Carnosine group there was no significant difference (P＞0.05).
2, lenticular opacity situation during medication was respectively organized in 4 days afterwards
According to the previous experiments method, experimental rat lenticular opacity situation during the observation medication was respectively organized in 4 days afterwards specifically sees Table 20.
Table 20 medication was respectively organized experimental rat lenticular opacity situation in 4 days relatively
Figure 2007101616010A00800191
Each test group result's statistical significance compares:
Matched group and model group P＜0.0005
Matched group and 5% L-Carnosine group P＜0.0005
Matched group and compositions B group P＜0.0005
Matched group and catalin group P＜0.0005
Model group and 5% L-Carnosine group P＜0.0005
Model group and compositions B group P＜0.0005
Model group and catalin group P＞0.05
5% L-Carnosine group and compositions B group P＜0.05
5% L-Carnosine group and catalin group P＜0.0005
Compositions B group and catalin group P＜0.0005
The statistical result of respectively organizing afterwards experimental rat lenticular opacity degree in 4 days by above-mentioned medication is as can be known:
Model group, 5% L-Carnosine group, compositions B group and catalin group lenticular opacity degree are apparently higher than matched group (P＜0.0005);
5% L-Carnosine group, compositions B group lenticular opacity degree are lower than respectively model group (P＜0.0005); Compositions B group lenticular opacity degree is lower than and 5% L-Carnosine group (P＜0.05).
3, lenticular opacity situation during medication was respectively organized in 6 days afterwards
According to the previous experiments method, experimental rat lenticular opacity situation during the observation medication was respectively organized in 6 days afterwards specifically sees Table 21.
Table 21 medication was respectively organized experimental rat lenticular opacity situation in 6 days relatively
Each test group result's statistical significance relatively
Matched group and model group P＜0.0005
Matched group and 5% L-Carnosine group P＜0.0005
Matched group and compositions B group P＜0.0005
Matched group and catalin group P＜0.0005
Model group and 5% L-Carnosine group P＜0.05
Model group and compositions B group P＜0.05
Model group and catalin group P＞0.05
5% L-Carnosine group and compositions B group P＞0.05
5% L-Carnosine group and catalin group P＜0.05
Compositions B group and catalin group P＜0.0005
The statistical result of respectively organizing experimental rat lenticular opacity degree in 6 days by above-mentioned medication is as can be known:
Model group, 5% L-Carnosine group, compositions B group and catalin group lenticular opacity degree are apparently higher than matched group (P＜0.0005);
5% L-Carnosine group, compositions B group lenticular opacity degree are lower than respectively model group (P＜0.05); Compositions B group lenticular opacity degree and 5% L-Carnosine group there was no significant difference (P＞0.05).
The above results shows: after the medication 2-6 days, each group of except matched group other is with the oxidative injury time lengthening, len's opacity increases the weight of gradually, and 5% L-Carnosine group and compositions B group lenticular opacity degree are starkly lower than other each experimental grouies, show that 5% L-Carnosine and compositions B have the effect that delays development of cataracts.
Test 2 compositions B for the impact of crystalline lens water-soluble protein content
1, the variation of crystalline lens water-soluble protein content during medication was respectively organized in 2 days afterwards
According to the previous experiments method, experimental rat crystalline lens water-soluble protein content situation of change during the observation medication was respectively organized in 2 days afterwards specifically sees Table 22.
The comparison (x+s) of crystalline lens water-soluble protein content was respectively organized in table 22 medication in 2 days
Figure 2007101616010A00800211
Each test group result's statistical significance compares:
Matched group and model group P＜0.005
Matched group and 5% L-Carnosine group P＜0.005
Matched group and compositions B group P＜0.005
Matched group and catalin group P＜0.005
Model group and 5% L-Carnosine group P＜0.0005
Model group and compositions B group P＜0.0005
Model group and catalin group P＞0.05
5% L-Carnosine group and compositions B group P＞0.05
5% L-Carnosine group and catalin group P＜0.001
Compositions B group and catalin group P＜0.0005
Medication was respectively organized experimental rat crystalline lens water-soluble protein content in 2 days and is learned by statistics processes and displays, model group, 5% L-Carnosine group, compositions B group and catalin group crystalline lens water-soluble protein content are starkly lower than matched group (P＜0.0005), 5% L-Carnosine group and compositions B group crystalline lens water-soluble protein content are higher than respectively model group (P＜0.0005), compositions B group crystalline lens water-soluble protein content and 5% L-Carnosine group there was no significant difference (P＞0.05).
2, the variation of crystalline lens water-soluble protein content during medication was respectively organized in 4 days afterwards
According to the previous experiments method, experimental rat crystalline lens water-soluble protein content situation of change during the observation medication was respectively organized in 4 days afterwards specifically sees Table 23.
The comparison (x+s) of crystalline lens water-soluble protein content was respectively organized in table 23 medication in 4 days
Figure 2007101616010A00800212
Each test group result's statistical significance compares:
Matched group and model group P＜0.005
Matched group and 5% L-Carnosine group P＜0.005
Matched group and compositions B group P＜0.005
Matched group and catalin group P＜0.005
Model group and 5% L-Carnosine group P＜0.05
Model group and compositions B group P＜0.005
Model group and catalin group P＞0.05
5% L-Carnosine group and compositions B group P＞0.05
5% L-Carnosine group and catalin group P＜0.05
Compositions B group and catalin group P＜0.005
Medication was respectively organized experimental rat crystalline lens water-soluble protein content in 4 days and is learned by statistics processes and displays, model group, 5% L-Carnosine group, compositions B group and catalin group crystalline lens water-soluble protein content are starkly lower than matched group (P＜0.0005), 5% L-Carnosine group and compositions B group crystalline lens water-soluble protein content are higher than respectively model group (P＜0.05, P＜0.005) and catalin group (P＜0.05, P＜0.005), compositions B group crystalline lens water-soluble protein content and 5% L-Carnosine group there was no significant difference (P＞0.05).
3, the variation of crystalline lens water-soluble protein content during medication was respectively organized in 6 days afterwards
According to the previous experiments method, experimental rat crystalline lens water-soluble protein content situation of change during the observation medication was respectively organized in 6 days afterwards specifically sees Table 24.
The comparison (x+s) of crystalline lens water-soluble protein content was respectively organized in table 24 medication in 6 days
Each test group result's statistical significance compares:
Matched group and model group P＜0.0005
Matched group and 5% L-Carnosine group P＜0.0005
Matched group and compositions B group P＜0.0005
Matched group and catalin group P＜0.0005
Model group and 5% L-Carnosine group P＜0.05
Model group and compositions B group P＜0.0005
Model group and catalin group P＞0.05
5% L-Carnosine group and compositions B group P＜0.0005
5% L-Carnosine group and catalin group P＜0.005
Compositions B group and catalin group P＜0.0005
Medication was respectively organized experimental rat crystalline lens water-soluble protein content in 6 days and is learned by statistics processes and displays, and model group, 5% L-Carnosine group, compositions B group and catalin group crystalline lens water-soluble protein content are starkly lower than matched group (P＜0.0005);
5% L-Carnosine group and compositions B group crystalline lens water-soluble protein content are higher than respectively model group (P＜0.05, P＜0.0005) and catalin group (P＜0.005, P＜0.0005);
Compositions B group crystalline lens water-soluble protein content is higher than 5% L-Carnosine (P＜0.0005).
The above results shows: after the medication 2-6 days, each organized other except matched group with the oxidative injury time lengthening, and the crystalline lens water-soluble protein content reduces gradually.
Lenticular water-solubility protein is lenticular structural protein, and is close with the maintain the relationship of the lenticular transparency.In the Cataractogenesis process, water-soluble protein content reduces.And 5% L-Carnosine group and compositions B group crystalline lens water-soluble protein content show that apparently higher than other each experimental grouies 5% L-Carnosine and compositions B have the effect that delays development of cataracts.
Test 3 compositions B to the impact of SOD activity in the crystalline lens
1, the variation of SOD activity in the crystalline lens was respectively organized in medication in 2 days afterwards
According to the previous experiments method, the situation of change of SOD activity in the experimental rat crystalline lens specifically saw Table 25 during the observation medication was respectively organized in 2 days afterwards.
The comparison (x+s) of crystalline lens SOD activity was respectively organized in table 25 medication in 2 days
Figure 2007101616010A00800231
Each test group result's statistical significance compares:
Matched group and model group P＜0.0005
Matched group and 5% L-Carnosine group P＜0.0005
Matched group and compositions B group P＜0.0005
Matched group and catalin group P＜0.0005
Model group and 5% L-Carnosine group P＜0.001
Model group and compositions B group P＜0.005
Model group and catalin group P＞0.05
5% L-Carnosine group and compositions B group P＞0.05
5% L-Carnosine group and catalin group P＜0.005
Compositions B group and catalin group P＜0.001
Medication was respectively organized experimental rat crystalline lens SOD activity in 2 days and is learned by statistics processes and displays:
Model group, 5% L-Carnosine group, compositions B group and catalin group crystal SOD activity are starkly lower than matched group (P＜0.0005);
5% L-Carnosine group and compositions B group crystal SOD activity are respectively apparently higher than model group (P＜0.001, P＜0.005) and catalin group (P＜0.005, P＜0.001); 5% L-Carnosine group crystal SOD is active to organize there was no significant difference (P＞0.05) with compositions B.
2, the variation of SOD activity in the crystalline lens was respectively organized in medication in 4 days afterwards
According to the previous experiments method, the situation of change of SOD activity in the experimental rat crystalline lens specifically saw Table 26 during the observation medication was respectively organized in 4 days afterwards.
The comparison (x+s) of crystalline lens SOD activity was respectively organized in table 26 medication in 4 days
Figure 2007101616010A00800241
Each test group result's statistical significance compares:
Matched group and model group P＜0.0005
Matched group and 5% L-Carnosine group P＜0.0005
Matched group and compositions B group P＜0.0005
Matched group and catalin group P＜0.0005
Model group and 5% L-Carnosine group P＜0.0005
Model group and compositions B group P＜0.0005
Model group and catalin group P＜0.0005
5% L-Carnosine group and compositions B group P＞0.05
5% L-Carnosine group and catalin group P＜0.01
Compositions B group and catalin group P＜0.05
Medication was respectively organized experimental rat crystalline lens SOD activity in 4 days and is learned by statistics processes and displays:
Model group, 5% L-Carnosine group, compositions B group and catalin group crystal SOD activity are starkly lower than matched group (P＜0.0005); 5% L-Carnosine group, compositions B group and catalin group crystal SOD activity are apparently higher than model group (P＜0.0005);
5% L-Carnosine group and compositions B group crystal SOD activity are higher than respectively catalin (P＜0.01, P＜0.05); 5% L-Carnosine group crystal SOD is active to organize there was no significant difference (P＞0.05) with compositions B.
3, the variation of SOD activity in the crystalline lens was respectively organized in medication in 6 days afterwards
According to the previous experiments method, the situation of change of SOD activity in the experimental rat crystalline lens specifically saw Table 27 during the observation medication was respectively organized in 6 days afterwards.
The comparison (x+s) of crystalline lens SOD activity was respectively organized in table 27 medication in 6 days
Figure 2007101616010A00800251
Each test group result's statistical significance compares:
Matched group and model group P＜0.0005
Matched group and 5% L-Carnosine group P＜0.0005
Matched group and compositions B group P＜0.0005
Matched group and catalin group P＜0.0005
Model group and 5% L-Carnosine group P＜0.0005
Model group and compositions B group P＜0.0005
Model group and catalin group P＜0.005
5% L-Carnosine group and compositions B group P＞0.05
5% L-Carnosine group and catalin group P＜0.05
Compositions B group and catalin group P＜0.05
Medication was respectively organized experimental rat crystalline lens SOD activity in 6 days and is learned by statistics processes and displays:
Model group, 5% L-Carnosine group, compositions B group and catalin group crystal SOD activity are starkly lower than matched group (P＜0.0005);
5% L-Carnosine group, compositions B group and catalin group crystal SOD activity are apparently higher than model group (P＜0.0005); 5% L-Carnosine group and compositions B group crystal SOD activity are higher than respectively catalin (P＜0.05); 5% L-Carnosine group crystal SOD is active to organize there was no significant difference (P＞0.05) with compositions B.
The above results shows: after the medication 2-6 days, each group of other except matched group was with the oxidative injury time lengthening, and crystalline lens SOD activity reduces gradually, points out lipid peroxide and radical pair crystalline lens activities of antioxidant enzymes inhibited.And 5% L-Carnosine group and compositions B group crystalline lens SOD activity show that apparently higher than other each experimental grouies external additional 5% L-Carnosine or compositions B can obviously improve the activity of crystalline lens antioxidase, delay the progress of rat cataract.
By above-mentioned test 1-3 as can be known:
1) after the medication 2-6 day, 5% L-Carnosine group and compositions B group lenticular opacity degree is starkly lower than other each experimental grouies (P＜0.05-0.0005), (P＜0.05-0.0005), crystalline lens SOD activity is apparently higher than other each experimental grouies (P＜0.05-0.0005) apparently higher than other each experimental grouies for the crystalline lens water-soluble protein content.Point out 5% L-Carnosine and compositions B can improve the activity of crystalline lens antioxidase, alleviate lenticular oxidative damage, the effect that delays development of cataracts is arranged.
2) after the medication 2-6 days, 5% L-Carnosine group lenticular opacity degree, crystalline lens water-soluble protein content and crystalline lens SOD active with compositions B group there was no significant difference (P＞0.05).
List of references
1. Sun Li, Jinsong ZHANG .DL-alpha-lipoic acid to rat experiment sugar from the inhibiting in vitro study of cataract. Recent Advances in Ophthalmology, 2004,24 (3): 197-200.
2. Peng De Xiang, Ding Zhuoping. the progress of Anti-Ageing Activity Peptides. modern food science and technology, 2006,22 (2): 267-270.
3. Guo Yong, Yan Hong. the effect that the carnosine prevention forms. eye Science, 2006,22 (2): 85-88.

Claims (2)

1. L-Carnosine pharmaceutical composition that is used for delaying development of cataracts, contain L-Carnosine 50g in its each kilogram of composition, phenoxyethanol 50g, polyvidone 10g, control pH value with boric acid-sodium carbonate between 6.8 to 8.0 as buffer salt, regulate osmotic pressure to 260-340mosm/L with an amount of potassium chloride, surplus is pure water, is prepared into according to a conventional method eye drop.
2. eye-gel preparation that is used for delaying development of cataracts, contain L-Carnosine 50g in its each kilogram eye-gel preparation, carbomer 934 is 24g, phenoxyethanol 50g, polyvidone 2g controls pH value with sodium hydrogen phosphate-sodium dihydrogen phosphate between 6.8 to 8.0 as buffer salt, regulates osmotic pressure to 260-340mosm/L with an amount of mannitol, surplus is pure water, is prepared into according to a conventional method gel for eye use.
CN 200710161601 2007-09-27 2007-09-27 Medicine composition containing L-carnosine for suspending the development of the cataract Active CN101396361B (en)

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