CN106619975A - Medicine injected at Zusanli acupoint for treating diabetic foot and preparation method of medicine - Google Patents
Medicine injected at Zusanli acupoint for treating diabetic foot and preparation method of medicine Download PDFInfo
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- CN106619975A CN106619975A CN201610904330.2A CN201610904330A CN106619975A CN 106619975 A CN106619975 A CN 106619975A CN 201610904330 A CN201610904330 A CN 201610904330A CN 106619975 A CN106619975 A CN 106619975A
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- extract
- reining
- lemahui
- zhusheye
- horse back
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- 210000001364 upper extremity Anatomy 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/80—Scrophulariaceae (Figwort family)
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Abstract
The invention belongs to the field of traditional Chinese medicines, and particularly relates to dicranostigma leptopodum injection and preparation method thereof. The dicranostigma leptopodum injection is prepared from a dicranostigma leptopodum extracting solution which is an active drug, and the dicranostigma leptopodum extracting solution is obtained by extracting through alcohol extraction and water precipitation, eluting and purifying through a resin column, and finally adding auxiliary materials. By the dicranostigma leptopodum injection, the blood glucose level of mice can be reduced remarkably, and further experiments show that by acupoint injection of the dicranostigma leptopodum injection, diabetic foot can be treated remarkably.
Description
Technical field
The present invention relates to LEMAHUI ZHUSHEYE and its preparation, and in particular to a kind of LEMAHUI ZHUSHEYE for treating diabetes
And preparation method thereof.
Background technology
Diabetes are a kind of serious complication of diabetes, are that diabetic is disabled, or even lethal major reason
One of, not only cause suffering to patient, and make it add huge financial burden.According to incompletely statistics, diabetic foot
Ulcer accounts for 20% or so.
Diabetic foot makes lower limb defencive function go down due to DPN, and big blood vessel and microangiopathies make artery
Hypoperfusion causes microcirculation disorder to be susceptible to diabetes.Wherein sensory nerve pathology merges too high mechanical stress, is to draw
Play the main initiating agent of foot ulcers and infection.Inflammation and histologic lesion are that a certain degree of alternate stress acts on a spy
The result in the fixed region for losing sensibility.From ground, shoes or other cause ulcer shape adjacent to the pressure or shearing force of toes
Into due to lacking normal Neuroprotective Mechanisms, ulcer often increases because of the presence of apophysis.Autonomic pathology is caused
Skin normally perspires regulatory function, skin temperature regulatory function and blood fortune regulating power and loses, and causes local organization pliability to drop
It is low, form the callosity of thickness and more broken and cracking.Additionally, the forfeiture for normally perspiring ability has blocked the water again of local organization
Change, cause to organize further destruction so that deep tissue is easier to bacteria planting.Morbidity of the kinesitherapy nerve pathology in diabetes
In also play certain effect, the contracture of foot inherence flesh causes typical claw-like microdactyly.The hyperextension of articulationes metatarsophalangeae is also proved to
Pressure under caput metatarsale can directly be increased so that the position is more likely formed ulcer.Nearly interphalangeal joint flexing is caused between the toe of projection
Joint dorsal part increases with the ulcerated risk in toe point plantar side, and vascular lesion causes the tissue of destruction to be difficult to heal.
The predominantly outer wiping ointment of medicament of diabetes is treated in prior art, local treatment is reached by promoting blood circulation, sterilization
The effect of diabetes, but the drug action of topical agent is slow, and cure the symptoms, not the disease, it is impossible to fundamentally cure diabetes
Foot.
The inventor of this patent has been surprisingly found that in research work, will rein in the horse back to be prepared into parenteral solution and carry out acupoint injection therapy and controls
Treat diabetes and taken unexpected effect, on this basis, inventor has been carried out after repeated multiple times research, is obtained
Technical scheme.
It is of the present invention to rein in the horse back as Scrophulariaceae bastard speedwell Pseudolysimachion linariifolium subsp.
The ground herb of dilatatum (Nakai & Kitagawa) D.Y.Hong.
The content of the invention
For the deficiencies in the prior art, the invention provides a kind of LEMAHUI ZHUSHEYE for treating diabetes and its preparation
Method, rapid reliable its liquid of its effect is directly injected into acupuncture point, and absorption process is very short, and without alimentary canal, not by pH,
Enzyme, food etc. affect, and without first pass effect, medicament contg is difficult loss, curative effect reliability.
To realize object above, the present invention is achieved by the following technical programs:
A kind of LEMAHUI ZHUSHEYE for treating diabetes, is made up of active medicine and excipient substance, and the active medicine is Le
Horse returns extract.
Preferably, the extract of reining in the horse back accounts for the 5% of parenteral solution cumulative volume.
Preferably, the excipient substance be sodium chloride, vitamin C, Sulfobutyl ether β _ cyclodextrin, glycerine, ethyl lactate,
One or more in citric acid, mannitol, glycerine, glycine, sorbierite.
A kind of preparation method of the LEMAHUI ZHUSHEYE for treating diabetes, step is as follows:
(1)It is in 65~85% ethanol water, to be warming up to 80~100 to take dry 3~8 times of amount concentration of addition of reining in the horse back
DEG C, cold filtration after the h of heating and refluxing extraction 0.5~2 obtains extract and the dregs of a decoction, and to the dregs of a decoction aforesaid operations 2 times are repeated, and merges
3 gained liquids, then vacuum distillation obtains alcohol extract;
(2)Distilled water is added in alcohol extract, after being well mixed, after standing 3~5 h, is filtered, take supernatant, then use milipore filter
Filter, filtrate concentration, it is extract at the beginning of 1.05~1.15 rein in the horse back to obtain relative density;
(3)Extract Jing D101B types macroporous absorbent resin at the beginning of reining in the horse back is processed, first with the pH 1~5 of 2~10 times of amounts of column volume
The aqueous solution eluted, collect eluent, then eluted with the 50% of 2~10 times of column volume amount ethanol, collect wash-out
Liquid, eluent merging twice, concentration obtains concentrate, then concentrate is adjusted into pH to 10~11 with alkali, at 60~80 DEG C
1~2h of heating, then pH to 3~5 is adjusted, then its Jing polyamide resin column is processed, with 2~5 times of water elution, collect wash-out
Liquid, then distills at 70~80 DEG C, removes ethanol extremely without alcohol taste, then adjusts pH to 6~7, obtains extract of reining in the horse back;
(4)Add sodium chloride and other drugs auxiliary material to be uniformly mixed in extract of reining in the horse back, be supplemented with water for injection extremely
Aequum, adjusts pH value 6.5~8.0, and it is filling then to rush nitrogen, sealing, with 100 DEG C of flowing steam sterilization 30 minutes, obtains final product
LEMAHUI ZHUSHEYE.
Preferably, the preparation method of the LEMAHUI ZHUSHEYE of the treatment diabetes, step is as follows:
(1)It is in 75% ethanol water, to be warming up to 85 DEG C to take dry 5 times of amount concentration of addition of reining in the horse back, is heated to reflux 1.5
Cold filtration after h, obtains extract and the dregs of a decoction, and to the dregs of a decoction aforesaid operations 2 times are repeated, and merges 3 gained liquids, and then decompression is steamed
Evaporate, obtain alcohol extract;
(2)Distilled water is added in alcohol extract, after being well mixed, after standing 4 h, is filtered, take supernatant, then use milipore filter mistake
Filter, filtrate concentration, it is extract at the beginning of 1.15 rein in the horse back to obtain relative density;
(3)Extract Jing D101B types macroporous absorbent resin at the beginning of reining in the horse back is processed, first with the water that the pH of 5 times of amounts of column volume is 3
Solution is eluted, and collects eluent, then is eluted with 50% ethanol of 5 times of amounts of column volume, collects eluent, is eluted twice
Liquid merges, concentration, obtains concentrate, then concentrate is adjusted into pH to 10 with ammoniacal liquor, and 1.5 h are heated at 60 DEG C, then uses salt
Then acid for adjusting pH is processed its Jing polyamide resin column to 5, with 3 times of water elution, eluent is collected, to washing at 80 DEG C
De- liquid is distilled, and removes ethanol extremely without alcohol taste, then adjusts pH to 7, obtains extract of reining in the horse back;
(4)Add sodium chloride and other drugs auxiliary material to be uniformly mixed in extract of reining in the horse back, be supplemented with water for injection extremely
Aequum, adjusts pH value 7, and it is filling then to rush nitrogen, sealing, with 100 DEG C of flowing steam sterilization 30 minutes, obtains final product re-injection of reining in the horse
Penetrate liquid.
Preferably, step(1)Described reduced pressure distillation process condition be -0.05~-0.09MPa, 40~90 DEG C.
Preferably, step(2)Milipore filter used by described ultrafiltration is 3000~6000 dalton, selected from diacetate fiber
Plain film, three cellulose acetate membrane, cyanoethyl cellulose film, PS membrane, sulfonated polysulfone membrane, poly (ether sulfone) film, sulfonated polyether sulfone
Film, Polysulfonamide, phenolphthalein side base polyarylsulfone (PAS) film, polyvinylidene fluoride film, polyacrylonitrile film, polyimide film, cellulose membrane, first
Base methyl acrylate-acrylonitrile-based copolymers membrane, polyacrylonitrile-cellulose diacetate blend film.
A kind of application of LEMAHUI ZHUSHEYE in treatment diabetes medicine is prepared, the LEMAHUI ZHUSHEYE is by active drug
Thing and excipient substance are constituted, and the active medicine is extract of reining in the horse back, accounts for the 5% of LEMAHUI ZHUSHEYE cumulative volume, the medicine
Auxiliary material is sodium chloride, glycerine, glycerine, water, and the preparation method of the LEMAHUI ZHUSHEYE, step is as follows:
(1)It is in 75% ethanol water, to be warming up to 85 DEG C by dry 5 times of amount concentration of addition of reining in the horse back, is heated to reflux 1.5
Cold filtration after h, obtains extract and the dregs of a decoction, and to the dregs of a decoction aforesaid operations 2 times are repeated, and merges 3 gained liquids, then technique bar
Part is -0.05~-0.09MPa, vacuum distillation under conditions of 40~90 DEG C, obtains alcohol extract;
(2)Distilled water is added in alcohol extract, after being well mixed, after standing 4 h, is filtered, take supernatant, then with 3000~6000
The cellulose diacetate ultrafiltration membrance filter of dalton, filtrate concentration, it is extract at the beginning of 1.15 rein in the horse back to obtain relative density;
(3)Extract Jing D101B types macroporous absorbent resin at the beginning of reining in the horse back is processed, first with the water that the pH of 5 times of amounts of column volume is 3
Solution is eluted, and collects eluent, then is eluted with 50% ethanol of 5 times of amounts of column volume, collects eluent, is eluted twice
Liquid merges, concentration, obtains concentrate, then concentrate is adjusted into pH to 10 with ammoniacal liquor, and 1.5 h are heated at 60 DEG C, then uses salt
Then acid for adjusting pH is processed its Jing polyamide resin column to 5, with 3 times of water elution, eluent is collected, to washing at 80 DEG C
De- liquid is distilled, and removes ethanol extremely without alcohol taste, then adjusts pH to 7, obtains extract of reining in the horse back;
(4)Add sodium chloride and other drugs auxiliary material to be uniformly mixed in extract of reining in the horse back, be supplemented with water for injection extremely
Aequum, adjusts pH value 7, and it is filling then to rush nitrogen, sealing, with 100 DEG C of flowing steam sterilization 30 minutes, obtains final product re-injection of reining in the horse
Penetrate liquid.
A kind of application of LEMAHUI ZHUSHEYE in medicament for treating insomnia is prepared, the LEMAHUI ZHUSHEYE by active medicine and
Excipient substance is constituted, and the active medicine is extract of reining in the horse back, accounts for the 5% of LEMAHUI ZHUSHEYE cumulative volume, the excipient substance
For sodium chloride, glycerine, glycerine, water, the preparation method of the LEMAHUI ZHUSHEYE, step is as follows:
(1)It is in 75% ethanol water, to be warming up to 85 DEG C by dry 5 times of amount concentration of addition of reining in the horse back, is heated to reflux 1.5
Cold filtration after h, obtains extract and the dregs of a decoction, and to the dregs of a decoction aforesaid operations 2 times are repeated, and merges 3 gained liquids, then technique bar
Part is -0.05~-0.09MPa, vacuum distillation under conditions of 40~90 DEG C, obtains alcohol extract;
(2)Distilled water is added in alcohol extract, after being well mixed, after standing 4 h, is filtered, take supernatant, then with 3000~6000
The cellulose diacetate ultrafiltration membrance filter of dalton, filtrate concentration, it is extract at the beginning of 1.15 rein in the horse back to obtain relative density;
(3)Extract Jing D101B types macroporous absorbent resin at the beginning of reining in the horse back is processed, first with the water that the pH of 5 times of amounts of column volume is 3
Solution is eluted, and collects eluent, then is eluted with 50% ethanol of 5 times of amounts of column volume, collects eluent, is eluted twice
Liquid merges, concentration, obtains concentrate, then concentrate is adjusted into pH to 10 with ammoniacal liquor, and 1.5 h are heated at 60 DEG C, then uses salt
Then acid for adjusting pH is processed its Jing polyamide resin column to 5, with 3 times of water elution, eluent is collected, to washing at 80 DEG C
De- liquid is distilled, and removes ethanol extremely without alcohol taste, then adjusts pH to 7, obtains extract of reining in the horse back;
(4)Add sodium chloride and other drugs auxiliary material to be uniformly mixed in extract of reining in the horse back, be supplemented with water for injection extremely
Aequum, adjusts pH value 7, and it is filling then to rush nitrogen, sealing, with 100 DEG C of flowing steam sterilization 30 minutes, obtains final product re-injection of reining in the horse
Penetrate liquid.
Specific embodiment
To make purpose, technical scheme and the advantage of the embodiment of the present invention clearer, below in conjunction with the embodiment of the present invention,
Technical scheme in the embodiment of the present invention is clearly and completely described, it is clear that described embodiment is the present invention one
Divide embodiment, rather than the embodiment of whole.Based on the embodiment in the present invention, those of ordinary skill in the art are not making
The every other embodiment obtained under the premise of creative work, belongs to the scope of protection of the invention.
Embodiment 1:
A kind of LEMAHUI ZHUSHEYE for treating diabetes, by account for parenteral solution cumulative volume 5% rein in the horse back extract and sodium chloride,
Glycine, glycine, sorbierite, citric acid, water composition.
A kind of preparation method of the LEMAHUI ZHUSHEYE for treating diabetes, step is as follows:
(1)Take the dry 100g that reins in the horse back, add 3~8 times of amount concentration be in 65~85% ethanol water be warming up to 80~
100 DEG C, cold filtration after the h of heating and refluxing extraction 0.5~2 obtains extract and the dregs of a decoction, and to the dregs of a decoction aforesaid operations 2 times are repeated,
Merge 3 gained liquids, then vacuum distillation obtains alcohol extract;
(2)Distilled water is added in alcohol extract, after being well mixed, after standing 3~5 h, is filtered, take supernatant, then use milipore filter
Filter, filtrate concentration, it is extract at the beginning of 1.05~1.15 rein in the horse back to obtain relative density;
(3)Extract Jing D101B types macroporous absorbent resin at the beginning of reining in the horse back is processed, first with the pH 1~5 of 2~10 times of amounts of column volume
The aqueous solution eluted, collect eluent, then eluted with the 50% of 2~10 times of column volume amount ethanol, collect wash-out
Liquid, eluent merging twice, concentration obtains concentrate, then concentrate is adjusted into pH to 10~11 with alkali, at 60~80 DEG C
1~2h of heating, then pH to 3~5 is adjusted, then its Jing polyamide resin column is processed, with 2~5 times of water elution, collect wash-out
Liquid, then distills at 70~80 DEG C, removes ethanol extremely without alcohol taste, then adjusts pH to 6~7, obtains extract of reining in the horse back;
(4)Add sodium chloride and other drugs auxiliary material to be uniformly mixed in extract of reining in the horse back, be supplemented with water for injection extremely
1000mL, adjusts pH value 6.5~8.0, and then sintered glass funnel filtration rushes nitrogen embedding in 2mL ampoules, and sealing uses 100
DEG C flowing steam sterilization 30 minutes, obtain final product LEMAHUI ZHUSHEYE.
Embodiment 2:
A kind of LEMAHUI ZHUSHEYE for treating diabetes, by account for parenteral solution cumulative volume 5% rein in the horse back extract and sodium chloride,
Sulfobutyl ether β _ cyclodextrin, sorbierite, ethyl lactate, water composition.
A kind of preparation method of the LEMAHUI ZHUSHEYE for treating diabetes, step is as follows:
(1)The dry 100g that reins in the horse back is taken, adds 6 times of amount concentration to be in 65% ethanol water, to be warming up to 80 DEG C, heated back
Cold filtration after stream extraction 0.5, obtains extract and the dregs of a decoction, and to the dregs of a decoction aforesaid operations 2 times are repeated, and merges 3 gained liquids, so
Vacuum distillation afterwards, obtains alcohol extract;
(2)Distilled water is added in alcohol extract, after being well mixed, after standing 3h, is filtered, take supernatant, then with ultrafiltration membrance filter,
Filtrate concentrates, and it is extract at the beginning of 1.05~1.15 rein in the horse back to obtain relative density;
(3)Extract Jing D101B types macroporous absorbent resin at the beginning of reining in the horse back is processed, first water-soluble with the pH 1 of 10 times of column volume amount
Liquid is eluted, and collects eluent, then is eluted with the 50% of 2 times of column volume amount ethanol, collects eluent, is eluted twice
Liquid merges, concentration, obtains concentrate, then concentrate is adjusted into pH to 11 with alkali, and 1h is heated at 80 DEG C, then adjusts pH to 5,
Then its Jing polyamide resin column is processed, with 5 times of water elution, collects eluent, then distilled at 80 DEG C, remove ethanol
Extremely without alcohol taste, then pH is adjusted to 6, obtain extract of reining in the horse back;
(4)Add sodium chloride and other drugs auxiliary material to be uniformly mixed in extract of reining in the horse back, be supplemented with water for injection extremely
1000mL, adjusts pH value 6.5~8.0, and then sintered glass funnel filtration rushes nitrogen embedding in 2mL ampoules, and sealing uses 100
DEG C flowing steam sterilization 30 minutes, obtain final product LEMAHUI ZHUSHEYE.
Embodiment 3:
A kind of LEMAHUI ZHUSHEYE for treating diabetes, by 5% rein in the horse back extract and sodium chloride, glycerine, water are constituted.
A kind of preparation method of the LEMAHUI ZHUSHEYE for treating diabetes, step is as follows:
(1)The dry 100g that reins in the horse back is taken, adds 5 times of amount concentration to be in 75% ethanol water, to be warming up to 85 DEG C, heated back
Cold filtration after 1.5 h is flowed, extract and the dregs of a decoction are obtained, aforesaid operations 2 times are repeated to the dregs of a decoction, merge 3 gained liquids, then
Vacuum distillation, obtains alcohol extract, the reduced pressure distillation process be -0.05 MPa, 60 DEG C;
(2)Distilled water is added in alcohol extract, after being well mixed, after standing 3 h, is filtered, take supernatant, then use 3000 dalton
The h of cyanoethyl cellulose membrane filtration 24, it is extract at the beginning of 1.15 rein in the horse back to obtain relative density;
(3)Extract Jing adsorption resin columns at the beginning of reining in the horse back are processed, and are first washed with the aqueous solution of the pH 3 of 5 times of amounts of column volume
It is de-, then eluted with the ethanol of 5 times of amounts of column volume, eluent is collected, concentrate is concentrated to give, then by concentrate ammoniacal liquor
PH to 10 is adjusted, 3 h are heated at 60 DEG C, then adjust pH to 5, then processed its Jing polyamide resin column, with 2 times of washing
It is de-, then distill at 80 DEG C, ethanol is removed extremely without alcohol taste, then pH is adjusted to 7, obtain extract of reining in the horse back;
(4)Add sodium chloride and other drugs auxiliary material to be uniformly mixed in extract of reining in the horse back, be supplemented with water for injection extremely
1000mL, adjusts pH value 7, and then sintered glass funnel filtration is rushed nitrogen embedding in 2mL ampoules, sealed, with 100 DEG C of stream
Logical steam sterilizing 30 minutes, obtains final product LEMAHUI ZHUSHEYE.
Embodiment 4:Rein in the horse back the research of extracting factor
1st, concentration of alcohol
According to the experimental design of extraction process, the ethanol of many variable concentrations carries out preferably, testing respectively with 25%, 50%, 75% and
95% ethanol as Extraction solvent, the rate of transform with extract protocatechuic acid as inspection target, in combination with protocatechuic acid
Content, different solvents are investigated.
Experimental technique:Weigh and rein in the horse back 5 parts, every part of 200g adds solvent to be heated to reflux 3 times, is heated to reflux temperature for 85
DEG C, the time is 1h, and the extract of three extractions is merged, and ethanol is reclaimed in vacuum distillation, and vacuum drying determines containing for protocatechuic acid
Amount(Assay method reference:Chen Yong, Fang Cuifen, Xiang Zhimin, Tang steps on peak .RP-HPLC and determines containing for capsule protocatechuic acid of reining in the horse back
Amount [J]. Chinese patent drug. 2007.29 (12):1867-1869. is similarly hereinafter), as a result such as table 1.
Wherein experimental group solvent is respectively the ethanol of 5 times of amounts 25%, 50%, 75% and 95%, and control group solvent is that 5 times of amounts are steamed
Distilled water.
Impact of the concentration of alcohol of table 1 to extraction process
As a result show, when being extracted with 75% ethanol, the content and the rate of transform of protocatechuic acid are above the ethanol of other concentration, because
The ethanol of this selection 75% is used as Extraction solvent.
, heating-up temperature
After Extraction solvent is determined, also need it is further preferred to Extracting temperature, test using 75% ethanol as Extraction solvent, carry
The time is taken for 1.5h, heating-up temperature be respectively 80 DEG C, 85 DEG C, 90 DEG C, 100 DEG C of rates of transform with extract protocatechuic acid be
Inspection target, is carried out preferably to the heat time, as a result such as table 2.
Impact of the heating-up temperature of table 2 to extraction process
As a result show, the rate of transform of the protocatechuic acid of experimental group 2 selects heating-up temperature to be 85 DEG C apparently higher than other groups.
, the heat time
After Extraction solvent is determined, also need it is further preferred to extraction time, test using 75% ethanol as Extraction solvent, plus
Hot reflux temperature is 85 DEG C, and extraction time is respectively the ethanol of 0.5h, 1h, 1.5h, 2h as Extraction solvent, with extract Central Plains
The rate of transform of catechuic acid is inspection target, the heat time is carried out preferably, as a result such as table 3.
Impact of the heat time of table 3 to extraction process
As a result show, the rate of transform of the protocatechuic acid of experimental group 3 is substantially poor with the thing of experimental group 4 higher than enforcement group 1 and experimental group 2
It is different, therefore select to be heated to reflux the time for 1.5h.
Embodiment 5:LEMAHUI ZHUSHEYE treats the experimental study of diabetes
1 experiment material
1.1 experimental animals
Experiment is from SPF levels health male Wistar rat 50(There is provided by Heilongjiang University of Chinese Medicine's animal center), body weight
For(250±10)g.Raise in 22~25 DEG C of room temperature, humidity 50%, draughty environment, with cleaning grade feed(By black dragon
River university of TCM animal center is provided)Single cage is fed, and is freely ingested, is drunk water.
1.2 reagents and medicine
Streptozotocin(Steptozotocin, STZ):Sigma(Sigma)Aldrich Shanghai trade Co., Ltd;
LEMAHUI ZHUSHEYE:LEMAHUI ZHUSHEYE is made up of active medicine and excipient substance, and the active medicine is carried to rein in the horse back
Liquid is taken, the 5% of LEMAHUI ZHUSHEYE cumulative volume is accounted for, the excipient substance is sodium chloride, glycerine, glycerine, water, the re-injection of reining in the horse
The preparation method of liquid is penetrated, step is as follows:
(1)By the dry 100g that reins in the horse back, add 5 times of amount concentration to be in 75% ethanol water, to be warming up to 85 DEG C, heat back
Cold filtration after 1.5 h is flowed, extract and the dregs of a decoction are obtained, aforesaid operations 2 times are repeated to the dregs of a decoction, merge 3 gained liquids, then
Process conditions are -0.05~-0.09MPa, vacuum distillation under conditions of 40~90 DEG C, obtain alcohol extract;
(2)Distilled water is added in alcohol extract, after being well mixed, after standing 4 h, is filtered, take supernatant, then with 3000~6000
The cellulose diacetate ultrafiltration membrance filter of dalton, filtrate concentration, it is extract at the beginning of 1.15 rein in the horse back to obtain relative density;
(3)Extract Jing D101B types macroporous absorbent resin at the beginning of reining in the horse back is processed, first with the water that the pH of 5 times of amounts of column volume is 3
Solution is eluted, and collects eluent, then is eluted with 50% ethanol of 5 times of amounts of column volume, collects eluent, is eluted twice
Liquid merges, concentration, obtains concentrate, then concentrate is adjusted into pH to 10 with ammoniacal liquor, and 1.5 h are heated at 60 DEG C, then uses salt
Then acid for adjusting pH is processed its Jing polyamide resin column to 5, with 3 times of water elution, eluent is collected, to washing at 80 DEG C
De- liquid is distilled, and removes ethanol extremely without alcohol taste, then adjusts pH to 7, obtains extract of reining in the horse back;
(4)Add sodium chloride and other drugs auxiliary material to be uniformly mixed in extract of reining in the horse back, be supplemented with water for injection extremely
1000mL, adjusts pH value 7, and then sintered glass funnel filtration is rushed nitrogen embedding in 2mL ampoules, sealed, with 100 DEG C of stream
Logical steam sterilizing 30 minutes, obtains final product LEMAHUI ZHUSHEYE.
1.3 prepare diabetes rat model
Randomly select 40 rats and diabetes rat model is set up using the method for STZ lumbar injections, it is former according to ischemia-reperfusion
Reason manufacture diabetes rat model(Ulcer of foot is one of cardinal symptom of diabetes).The right side to diabetic model group rat
The same position of hind leg is ligatured, and using 10 extra section suture ligature rat ankle and foots, 2h is ligatured every time, decontrols 30min, daily
Repeat ligation 3 times, 3d is ligatured altogether, operate 1 time every 1d.There is skin with ligated limbs to black, bubble, bleeding blister, ulcer are true
Recognize the successful standard of modeling.During ligation, operated by same person, it is ensured that identical using strength during ligation.It is all ligated
The equal Cheng Mo of rat.
Intervention of 1.4 LEMAHUI ZHUSHEYEs to diabetes rat model
40 diabetes rat models are numbered according to body weight, according to random digits table, by diabetes rat model point
For 2 groups, 20 per group, respectively:LEMAHUI ZHUSHEYE treatment group and diabetes blank control group.To LEMAHUI ZHUSHEYE amount
Group is according to 300mg/(kg·d)(Equivalent to 5 times of clinical patients consumption per day)Dosage, to mouse left side gastrocnemius position note
Penetrate medicine, 1 time/d.To diabetes blank control group use physiological saline gavage, 2 times/d.Body weight is measured weekly, changes medication
Amount, shares medicine 6 weeks.The fasting blood-glucose of each group rat is detected respectively(FBG), Diagnostic Value of Fasting Serum insulin(FSI), lectin from hemolymph(Including
High shear rate, low shear rate, Plasma Viscosity, hematocrit, erythrocyte aggregation index, 6 indexs of deformable index).
1.5 statistical methods
Database is set up with Excel, with SPSS13.0 statistical analysis software analyze datas.Measurement data with± s represents, two groups
Between compare and checked using t, multigroup is compared and adopts variance analysis;Enumeration data compares and adopts χ2Inspection, inspection level α=0.05.
2 results
2.1 LEMAHUI ZHUSHEYE treatment groups, diabetes blank control group comparitive study
After the LEMAHUI ZHUSHEYEs of 6 weeks are intervened, after the treatment of diabetes LEMAHUI ZHUSHEYE treatment group, foot symptom is obvious
Take a turn for the better;And the body weight of diabetes blank control group rat is decreased obviously before relatively treating, foot symptom aggravates.It is shown in Table 4.
The LEMAHUI ZHUSHEYE treatment group of table 4 and diabetes blank control group foot symptom score level condition(Only, %)
Note:Two groups are compared, χ2=10.489, P=0.015.Two groups have notable difference.
Each Testing index compares between 2.2 LEMAHUI ZHUSHEYE treatment groups and diabetes blank control group
Before the experiment initial stage treats, the FBG of two groups of rats, body weight and foot symptom score no significant difference;Rein in the horse re-injection
After the treatment FBG is significantly reduced for She Ye treatment groups, and FSI is significantly raised, high shear rate in lectin from hemolymph, low shear rate, Plasma Viscosity,
Hematocrit, erythrocyte aggregation index, deformable index work property is reduced.It is shown in Table 5.
Body weight, FBG, FSI and blood flow after the LEMAHUI ZHUSHEYE treatment group of table 5 and the treatment of diabetes blank control group
The comparison of change(±s)
3 conclusions
This experiment confirms:(1)LEMAHUI ZHUSHEYE can improve the fasting blood-glucose of diabetes B rat and the disease of foot ulcers
Shape, with certain effect for reducing blood sugar, treatment diabetes.(2)LEMAHUI ZHUSHEYE can significantly reduce diabetes
6 indexs, have improvement result to Hemorheology in rat lectin from hemolymph.This experiment also demonstrates that LEMAHUI ZHUSHEYE has and reduces
The blood sugar of diabetes rat, illustrates that the medicine is influential on diabetes;The serum insulin level of rat is in medication in experiment
After have significantly rising, illustrate that this medicine can improve the islet function of diabetes B rat of induction.
Embodiment 6:The clinical observation of 0 grade of diabetes of LEMAHUI ZHUSHEYE type Treated with Point-injection Therapy
Diabetes make the quality of life degradation of patient, and treat difficult, and treatment cycle is long, and medical expense is high, to trouble
Person and society bring white elephant.Inventor is set with the artificial research object of 0 grade of diabetic foot using random packet control
The research method of meter, gives different treatment and nursing measures, and the deterioration intervention to 0 grade of diabetes is studied, so as to explore
A kind of more satisfactory treatment and nursing method of 0 grade of diabetes, prevents ulcer of foot from occurring.
1 object and method
1.1 objects
0 grade of the diabetes B diabetes 24 in December, 2015 to the hospitalization of in July, 2016, patient with diabetic feet is accorded with
Close the diagnostic criteria of diabetes world clinical guidelines.It is wherein male 12, female 12,50 years old~65 years old age, averagely(62.9±
50.1)Year, diabetic duration(3.6±2.1)Year;Receive diabetes systematic treating, OHA or insulin therapy have
Effect.All patients feet's skins are complete, and without open focus, Jing Chinese medical discriminations are venation block, show as pedal skin purplish red
Or it is purple dim, limbs send out cool numbness, sharp ache, fuyang pulse decrease.It is randomly divided into experimental group and control group, per group 12, two groups
Patient's ordinary circumstance(Sex, the age, the course of disease, symptom, sign, blood sugar, HbAle egg from, blood fat, blood pressure etc.)Without statistics
Meaning(P>0.05), with comparativity.
1.2 methods
Related patient is pressed into the sequencing numbering of institute, two groups are incorporated at random respectively.
Control group:Diabetic diet and exercise therapy add OHA and(Or)Insulin therapy controls blood sugar, makes sky
Abdomen blood sugar≤7.0mmoL/L, postprandial blood sugar≤10.0mmoL/L, using conventional foodcare and health education.
Experimental group:LEMAHUI ZHUSHEYE acupoint injection therapy is adopted on the basis of former equal treatment and nursing.With reference to National Technical prison
Superintend and direct People's Republic of China's standard of office's promulgation《Location of acupoints》, determine point st 36.Patient makes even clinostatism, and a parapodum three is taken every time
In acupuncture point, left and right wheels are for using, after cave area skin routine disinfection, extracted with the 2mL sterilizing syringes for being furnished with No. 5 syringe needles and reined in the horse back
Parenteral solution 2mL, straight thrust is subcutaneous 0.5 11 cun, and pumpback after bringing about the desired sensation is injected into re-injection of reining in the horse and penetrates without blood and impassivity Rediating Pain per cave
Liquid 1mL, one time a day, and 15 times is a course for the treatment of, treats a course for the treatment of.
LEMAHUI ZHUSHEYE:LEMAHUI ZHUSHEYE is made up of active medicine and excipient substance, and the active medicine is to rein in the horse
Extract is returned, the 5% of LEMAHUI ZHUSHEYE cumulative volume is accounted for, the excipient substance is sodium chloride, glycerine, glycerine, water, and this is reined in the horse
The preparation method of parenteral solution is returned, step is as follows:
(1)By the dry 100g that reins in the horse back, add 5 times of amount concentration to be in 75% ethanol water, to be warming up to 85 DEG C, heat back
Cold filtration after 1.5 h is flowed, extract and the dregs of a decoction are obtained, aforesaid operations 2 times are repeated to the dregs of a decoction, merge 3 gained liquids, then
Process conditions are -0.05~-0.09MPa, vacuum distillation under conditions of 40~90 DEG C, obtain alcohol extract;
(2)Distilled water is added in alcohol extract, after being well mixed, after standing 4 h, is filtered, take supernatant, then with 3000~6000
The cellulose diacetate ultrafiltration membrance filter of dalton, filtrate concentration, it is extract at the beginning of 1.15 rein in the horse back to obtain relative density;
(3)Extract Jing D101B types macroporous absorbent resin at the beginning of reining in the horse back is processed, first with the water that the pH of 5 times of amounts of column volume is 3
Solution is eluted, and collects eluent, then is eluted with 50% ethanol of 5 times of amounts of column volume, collects eluent, is eluted twice
Liquid merges, concentration, obtains concentrate, then concentrate is adjusted into pH to 10 with ammoniacal liquor, and 1.5 h are heated at 60 DEG C, then uses salt
Then acid for adjusting pH is processed its Jing polyamide resin column to 5, with 3 times of water elution, eluent is collected, to washing at 80 DEG C
De- liquid is distilled, and removes ethanol extremely without alcohol taste, then adjusts pH to 7, obtains extract of reining in the horse back;
(4)Add sodium chloride and other drugs auxiliary material to be uniformly mixed in extract of reining in the horse back, be supplemented with water for injection extremely
1000mL, adjusts pH value 7, and then sintered glass funnel filtration is rushed nitrogen embedding in 2mL ampoules, sealed, with 100 DEG C of stream
Logical steam sterilizing 30 minutes, obtains final product LEMAHUI ZHUSHEYE.
1.3 clinical observation on the therapeutic effect
1.3.1 judgment criteria
With reference to 2002《New Chinese medicine guideline of clinical investigations》In《New Chinese medicine physiotherapy diabete(Diabetes)Clinic
Will be with regard to guideline》.
Recovery from illness:Pain, numbness, the symptom treatment such as send out cool after all disappear, skin color recovers normal;
It is effective:Pain, numbness, the symptom treatment such as send out cool after be obviously improved, skin color is obviously improved;
Effectively:Pain, numbness, the symptom treatment such as send out cool after take a turn for the better, skin color improves;
It is invalid:Pain, numbness, the symptom treatment such as send out cool after change not substantially, skin color does not improve.
All objects are responsible for observing related symptoms and sign and recording by special messenger(Observer is not involved in treatment, does not also know point
Group situation), observe and be detected on treatment the previous day for the first time and last time is treated and carried out one day after.
1.3.2 school work rheology Hygienic monitoring on hands of childhood
The arteria dorsalis pedis hemorheology ginseng before and after Case treatment is determined using Acuson28XP/10 type color Doppler echocardiographies instrument
Number, is carried out one day after in treatment the previous day first time and last time treatment.
1.4 side reactions are observed
After injection 0.5h, 2h, 6h, 12h, 24h, three days, seven days, observation whether there is the local reactions such as itching, redness, pain
And the general reaction such as have a fainting spell during acupuncture treatment, have a headache, suffering a shock.
1.5 statistical procedures
All market demand SPSS11.0 statistical softwares are processed, and using inspection, measurement data is with for the inspection of enumeration data and rate
Number ± standard deviation()Represent, checked using t.
2 results
2.1 two groups of patient's biped therapeutic effects compare
6 two groups of patient's biped therapeutic effects of table compare
The result of table 6 shows:The treatment total effective rate of experimental group is 100.00%, and the treatment total effective rate of control group is 58.33%, two
Compare that there were significant differences between group(=12.596, * P<0.01).
2.2 two groups of patient blood rheological parameters compare
7 two groups of patient's arteria dorsalis pedis blood rheology parameters of table compare()
The result of table 7 shows:The front instep Diameter of two groups of Case treatments, VPV and CBF numeric ratio are compared with without statistics
Learn difference(P>0.05).2. itself compare before and after two groups of Case treatments, instep Diameter expands after treatment group's Case treatment, blood
Flow velocity degree is accelerated, and CBF increases, compares with statistical significance with before treatment(P<0.01), and become after control group Case treatment
Change not statistically significant(P>0.05).3. compare after two groups of Case treatments, each Experiment Parameter group is superior to control group, with notable
Statistical significance (P<0.01).
3 conclusions
After intervening two groups of diabetes Bs, 0 grade of diabetes difference treatment and nursing, clinical efficacy and arteria dorsalis pedis blood stream
Become correlation study to show:Experimental group is substantially better than control group, is more beneficial for improving pedal blood circulation, alleviates diabetes
Foot symptom and sign, desirably prevent the generation of ulcer of foot.
Diabetes are one of diabetes important complications, and according to external investigation, about 15% diabetic will occur foot
Ulcer, more than 80% diabetic cuts toe or amputation due to ulcer of foot, so if can rationally control 0 phase diabetes
Treat and carry out nursing and interfering in time, prevent the generation of ulcer of foot, patient suffering will be reduced, mitigate burden on society, this exactly traditional Chinese medical science
The preferably embodiment of " not treating the disease affected, preventive treatment of disease " theory.The pathogenesis of diabetes is related to DPN, vascular lesion and sense
Dye, and blood circulation disorder is the basis of diabetes morbidity.Thus on Primary Care, using the treatment side of tcm characteristic
Method, catches blood circulatory disorder this pathologic process, and being reached by promoting blood circulationization addiction medicine acupoint injection therapy improves blood circulation, eliminates induction
Factor, so as to intervene the deterioration of 0 grade of diabetes, prevents ulcer of foot from occurring.
This result of study shows, to 0 grade of diabetes of diabetes B using LEMAHUI ZHUSHEYE in Zusanli, Sanyinjiao
Acupoint injection therapy, can promote 0 grade of diabetes remission, prevent the generation of ulcer of foot, and patient has no adverse reaction, and is a kind of peace
Entirely, 0 grade of effective, simple diabetes intervening measure.
Embodiment 7:Therapeutic action of the LEMAHUI ZHUSHEYE to Diabetic Nephropathy is studied
1 materials and methods
1.1 animals, medicine, reagent and instrument
Cleaning grade Wistar rats 45, purchased from Heilongjiang University of Chinese Medicine's Experimental Animal Center.
LEMAHUI ZHUSHEYE:LEMAHUI ZHUSHEYE is made up of active medicine and excipient substance, and active medicine is carried to rein in the horse back
Liquid is taken, the 5% of LEMAHUI ZHUSHEYE cumulative volume is accounted for, excipient substance is sodium chloride, glycerine, glycerine, water, the LEMAHUI ZHUSHEYE
Preparation method, step is as follows:
(1)By the dry 100g that reins in the horse back, add 5 times of amount concentration to be in 75% ethanol water, to be warming up to 85 DEG C, heat back
Cold filtration after 1.5 h is flowed, extract and the dregs of a decoction are obtained, aforesaid operations 2 times are repeated to the dregs of a decoction, merge 3 gained liquids, then
Process conditions are -0.05~-0.09MPa, vacuum distillation under conditions of 40~90 DEG C, obtain alcohol extract;
(2)Distilled water is added in alcohol extract, after being well mixed, after standing 4 h, is filtered, take supernatant, then with 3000~6000
The cellulose diacetate ultrafiltration membrance filter of dalton, filtrate concentration, it is extract at the beginning of 1.15 rein in the horse back to obtain relative density;
(3)Extract Jing D101B types macroporous absorbent resin at the beginning of reining in the horse back is processed, first with the water that the pH of 5 times of amounts of column volume is 3
Solution is eluted, and collects eluent, then is eluted with 50% ethanol of 5 times of amounts of column volume, collects eluent, is eluted twice
Liquid merges, concentration, obtains concentrate, then concentrate is adjusted into pH to 10 with ammoniacal liquor, and 1.5 h are heated at 60 DEG C, then uses salt
Then acid for adjusting pH is processed its Jing polyamide resin column to 5, with 3 times of water elution, eluent is collected, to washing at 80 DEG C
De- liquid is distilled, and removes ethanol extremely without alcohol taste, then adjusts pH to 7, obtains extract of reining in the horse back;
(4)Add sodium chloride and other drugs auxiliary material to be uniformly mixed in extract of reining in the horse back, be supplemented with water for injection extremely
1000mL, adjusts pH value 6.5~8.0, and then sintered glass funnel filtration rushes nitrogen embedding in 2mL ampoules, and sealing uses 100
DEG C flowing steam sterilization 30 minutes, obtain final product LEMAHUI ZHUSHEYE.
The mono- grand antibody of sheep anti mouse CTGF, purchased from CellSignaling companies of the U.S.;Sheep anti mouse β-actin antibody, is purchased from
Sigma companies;Streptozotocin, purchased from Sigma companies.Small table centrifuge(Mierofuge18), purchased from Bechman
Coulter companies;Turbula shaker(Vortex-GenieZ), purchased from Scilnd companies of the U.S.;Automatic clinical chemistry analyzer, purchase
From Beckman Coulter Inc. of the U.S..
The packet of 1.2 rats, modelling and administration
45 cleaning grade Wistar rats are randomly divided into normal group, control group and experimental group, each 15.Control group and experimental group
Give Streptozotocin 55mg/kg intraperitoneal injections and set up Diabetic nephropathy animal model, normal group gives the life of Isodose
Reason salt solution lumbar injection.After modeling success, experimental group rat gives LEMAHUI ZHUSHEYE 300mg/kg intraperitoneal injections, 1 time/d;
Normal group and control group give the physiological saline intraperitoneal injection of Isodose, 1 time/d.Three groups of rat free water feeds, no
Give any hypoglycemic to process.
The detection of 1.3 blood sugar, twenty-four-hour urine albumen, serum creatinine and urea nitrogen
Before three groups of rat injections(Before treatment)And when injecting 12 weeks(After treatment)Its blood sugar, twenty-four-hour urine albumen, blood are detected respectively
Clear creatinine and urea nitrogen.
1.4 statistical methods
Using the statistical softwares of Stata 10.0.Measurement data is used± s represents, three groups are compared and adopt variance analysis, between group two-by-two
Relatively checked using t, correlation analysis Pearson relevant function methods.P < 0.05 are that difference is statistically significant.
2 results
Three groups of rat blood sugars, twenty-four-hour urine albumen, serum creatinine and urea nitrogen levels compare before and after 2.1 treatments
Experimental group and control rats twenty-four-hour urine albumen, serum creatinine and urea nitrogen levels are higher than normal group before treatment(The equal < of P
0.05);Without substantially reduction before experimental group Rat 24 h Urine proteins, serum creatinine and urea nitrogen are relatively treated after treatment(The equal > of P
0.05).Refer to table 8.
Blood sugar, twenty-four-hour urine albumen, serum creatinine and urea nitrogen levels compare before and after 8 three groups of rat treatments of table(±s)
3 discuss and conclusion
After LEMAHUI ZHUSHEYE 12 weeks, its 24 hours Urine proteins, serum creatinine and urea nitrogen levels are not apparent from reducing, and
After being not below treatment of control group, illustrate that LEMAHUI ZHUSHEYE can not reduce diabetes rat albuminuria, Renal Function in Rats is not had
Protective effect, it is impossible to treat diabetic nephropathy.
Embodiment 8:The experimental study of LEMAHUI ZHUSHEYE treatment insomnia
1 experiment material
Medicine:
LEMAHUI ZHUSHEYE:LEMAHUI ZHUSHEYE:LEMAHUI ZHUSHEYE is made up of active medicine and excipient substance, and active medicine is
Rein in the horse back extract, account for the 5% of LEMAHUI ZHUSHEYE cumulative volume, excipient substance is sodium chloride, glycerine, glycerine, water, and this is reined in the horse
The preparation method of parenteral solution is returned, step is as follows:
(1)By the dry 100g that reins in the horse back, add 5 times of amount concentration to be in 75% ethanol water, to be warming up to 85 DEG C, heat back
Cold filtration after 1.5 h is flowed, extract and the dregs of a decoction are obtained, aforesaid operations 2 times are repeated to the dregs of a decoction, merge 3 gained liquids, then
Process conditions are -0.05~-0.09MPa, vacuum distillation under conditions of 40~90 DEG C, obtain alcohol extract;
(2)Distilled water is added in alcohol extract, after being well mixed, after standing 4 h, is filtered, take supernatant, then with 3000~6000
The cellulose diacetate ultrafiltration membrance filter of dalton, filtrate concentration, it is extract at the beginning of 1.15 rein in the horse back to obtain relative density;
(3)Extract Jing D101B types macroporous absorbent resin at the beginning of reining in the horse back is processed, first with the water that the pH of 5 times of amounts of column volume is 3
Solution is eluted, and collects eluent, then is eluted with 50% ethanol of 5 times of amounts of column volume, collects eluent, is eluted twice
Liquid merges, concentration, obtains concentrate, then concentrate is adjusted into pH to 10 with ammoniacal liquor, and 1.5 h are heated at 60 DEG C, then uses salt
Then acid for adjusting pH is processed its Jing polyamide resin column to 5, with 3 times of water elution, eluent is collected, to washing at 80 DEG C
De- liquid is distilled, and removes ethanol extremely without alcohol taste, then adjusts pH to 7, obtains extract of reining in the horse back;
(4)Add sodium chloride and other drugs auxiliary material to be uniformly mixed in extract of reining in the horse back, be supplemented with water for injection extremely
1000mL, adjusts pH value 6.5~8.0, and then sintered glass funnel filtration rushes nitrogen embedding in 2mL ampoules, and sealing uses 100
DEG C flowing steam sterilization 30 minutes, obtain final product LEMAHUI ZHUSHEYE.
Yellow Jackets:Beijing land for growing field crops Feng Tuo chemical technologies Co., Ltd produces;
Strychnine nitrate parenteral solution:Beijing Double-Crane Pharmaceutical Co., Ltd produces;
Diazepam:Tianjin KingYork Amino Acid Co., Ltd. produces;
Animal:Kunming mouse, body weight 18g~22g, female dual-purpose is provided by Heilongjiang University of Chinese Medicine experimental center.
2 experimental techniques and result
The research of 2.1 pairs of spontaneous activity in mice
Mouse 30, male and female are regardless of, and are randomly divided into 3 groups, i.e. saline control group, LEMAHUI ZHUSHEYE group, and it is positive right to stabilize
According to group, 10 per group.
After adapting to 5 minutes in mouse input animal activity case before administration, animal walking time and the forelimb of 2 minutes are recorded
The numerical value of number of times is praised upwards as normal value.
Then give respectively:
LEMAHUI ZHUSHEYE group:Injected LEMAHUI ZHUSHEYE 1mL, lumbar injection.
Stable positive controls:Injected diazepam 1mL, lumbar injection;
Saline control group:Injected 0.9% sodium chloride injection of same volume, lumbar injection.
Each group mouse determines mouse walking time and double forelimbs praises upwards number of times by the same method 60 minutes after administration.Knot
Fruit with± S represents, the results are shown in Table 9.
Impact of the table 9 to spontaneous activity in mice number of times(± S, n=10)
Note:Compare with saline control group, * P< 0.01;Compare with stable group:# P > 0.05;
Result shows in table:LEMAHUI ZHUSHEYE has obvious sedation to mouse, and comparing with saline control group has
Significant difference(P < 0.01).LEMAHUI ZHUSHEYE group sedation substantially, with stable group indifference is compared(P> 0.05).Knot
Fruit shows:LEMAHUI ZHUSHEYE has obvious sedation.
2.2 researchs that mice sleep is affected
2.2.1 to the impact of the mouse yellow Jackets length of one's sleep
Mouse 30, male and female are regardless of, and are randomly divided into 3 groups, i.e. saline control group, LEMAHUI ZHUSHEYE group, and it is positive right to stabilize
According to group, 10 per group.
Then give respectively:
LEMAHUI ZHUSHEYE group:Injected LEMAHUI ZHUSHEYE 1mL, lumbar injection;
Stable positive controls:Injected diazepam 1mL, lumbar injection;
Saline control group:Injected 0.9% sodium chloride injection of same volume, lumbar injection.
After administration 60 minutes, every group of mouse gives respectively yellow Jackets 40mg/kg, lumbar injection.Record sleeping for mouse
The dormancy time(It is sleep time from righting reflex loss to recovery time with righting reflex loss as time for falling asleep).As a result
It is shown in Table 10.
Impact of the table 10 to the mouse yellow Jackets length of one's sleep(± S, n=10)
2.2.2 to the impact of mouse yellow Jackets sub-threshold dose
Mouse 30, male and female are regardless of, and are randomly divided into 3 groups, i.e. saline control group, LEMAHUI ZHUSHEYE group, and it is positive right to stabilize
According to group, 10 per group.
Then give respectively:
LEMAHUI ZHUSHEYE group:Injected LEMAHUI ZHUSHEYE 1mL, lumbar injection;
Stable positive controls:Injected diazepam 1mL, lumbar injection;
Saline control group:Injected 0.9% sodium chloride injection of same volume, lumbar injection.
After administration 30 minutes, each group mouse gives respectively yellow Jackets 30mg/kg, and lumbar injection is observed and is recorded into
Sleep number of animals(In 15 minutes mouse righting reflex loss up to the mouse number on 1 minute as sleep number of animals), calculate sleep rate.Knot
Fruit is shown in Table 11.
Impact of the table 11 to mouse pentobarbital sodium sub-threshold lull dosage
The above two is tested and illustrate LEMAHUI ZHUSHEYE and have obvious syngignoscism to mouse, with saline control group ratio
Compared with there were significant differences(P < 0.01).LEMAHUI ZHUSHEYE group has obvious syngignoscism.
The research of 2.3 anti-strychnine nitrate convulsive attack effects
Mouse 30, male and female are regardless of, and are randomly divided into 3 groups, i.e. saline control group, LEMAHUI ZHUSHEYE group, and it is positive right to stabilize
According to group, 10 per group.
Then give respectively:
LEMAHUI ZHUSHEYE group:Injected LEMAHUI ZHUSHEYE 1mL, lumbar injection.
Stable positive controls:Injected diazepam 1mL, lumbar injection;
Saline control group:Injected 0.9% sodium chloride injection of same volume, lumbar injection.
Each group mouse gives respectively strychnine nitrate parenteral solution 1.5mg/kg, hypodermic injection, note after administration 60 minutes
Record mice convulsion number(There is generalized tonic with mouse to faint from fear as index).The results are shown in Table 12.
The effect of the anti-strychnine nitrate convulsive attack of table 12(n= 10)
Note:Compare with saline control group:*P < 0.01;Compare with stable group:※P > 0.05
As a result show, stable group plays the role of to resist strychnine nitrate convulsive attack, each group and life with LEMAHUI ZHUSHEYE group
Reason saline control group compares, and is statistically analyzed the equal significance of difference.And the anticonvulsant action of discovery LEMAHUI ZHUSHEYE
Most strong, curative effect preferably, illustrates that LEMAHUI ZHUSHEYE has anticonvulsant effect.
3 conclusions
The study find that, LEMAHUI ZHUSHEYE can suppress spontaneous activity in mice;Impact to mice sleep has with yellow Jackets
Synergy;Play the role of to resist strychnine nitrate convulsive attack.Illustrate that LEMAHUI ZHUSHEYE has obvious calmness, hypnosis, resists
Convulsion effect, the prospect with exploitation medicament for treating insomnia.
It should be noted that herein, such as first and second or the like relational terms are used merely to a reality
Body or operation make a distinction with another entity or operation, and not necessarily require or imply these entities or deposit between operating
In any this actual relation or order.And, term " including ", "comprising" or its any other variant are intended to
Nonexcludability is included, so that a series of process, method, article or equipment including key elements not only will including those
Element, but also including other key elements being not expressly set out, or also include for this process, method, article or equipment
Intrinsic key element.In the absence of more restrictions, the key element for being limited by sentence "including a ...", it is not excluded that
Also there is other identical element in process, method, article or equipment including the key element.
Above example only to illustrate technical scheme, rather than a limitation;Although with reference to the foregoing embodiments
The present invention has been described in detail, it will be understood by those within the art that:It still can be to aforementioned each enforcement
Technical scheme described in example is modified, or carries out equivalent to which part technical characteristic;And these modification or
Replace, the essence for not making appropriate technical solution departs from the present invention.
Claims (10)
1. a kind of LEMAHUI ZHUSHEYE for treating diabetes, it is characterised in that be made up of active medicine and excipient substance, it is described
Active medicine is extract of reining in the horse back.
2. the LEMAHUI ZHUSHEYE of diabetes is treated as claimed in claim 1, it is characterised in that the extract of reining in the horse back
Account for the 5% of parenteral solution cumulative volume.
3. the LEMAHUI ZHUSHEYE of diabetes is treated as claimed in claim 1, it is characterised in that the excipient substance is chlorine
Change sodium, vitamin C, Sulfobutyl ether β _ cyclodextrin, glycerine, ethyl lactate, citric acid, mannitol, glycerine, glycine, sorbierite
In one or more.
4. it is a kind of as described in claims 1 to 3 is arbitrary treatment diabetes LEMAHUI ZHUSHEYE preparation method, its feature
It is that step is as follows:
(1)It is in 65~85% ethanol water, to be warming up to 80~100 to take dry 3~8 times of amount concentration of addition of reining in the horse back
DEG C, cold filtration after the h of heating and refluxing extraction 0.5~2 obtains extract and the dregs of a decoction, and to the dregs of a decoction aforesaid operations 2 times are repeated, and merges
3 gained liquids, then vacuum distillation obtains alcohol extract;
(2)Distilled water is added in alcohol extract, after being well mixed, after standing 3~5 h, is filtered, take supernatant, then use milipore filter
Filter, filtrate concentration, it is extract at the beginning of 1.05~1.15 rein in the horse back to obtain relative density;
(3)Extract Jing D101B types macroporous absorbent resin at the beginning of reining in the horse back is processed, first with the pH 1~5 of 2~10 times of amounts of column volume
The aqueous solution eluted, collect eluent, then eluted with the 50% of 2~10 times of column volume amount ethanol, collect wash-out
Liquid, eluent merging twice, concentration obtains concentrate, then concentrate is adjusted into pH to 10~11 with alkali, at 60~80 DEG C
1~2h of heating, then pH to 3~5 is adjusted, then its Jing polyamide resin column is processed, with 2~5 times of water elution, collect wash-out
Liquid, then distills at 70~80 DEG C, removes ethanol extremely without alcohol taste, then adjusts pH to 6~7, obtains extract of reining in the horse back;
(4)Add sodium chloride and other drugs auxiliary material to be uniformly mixed in extract of reining in the horse back, be supplemented with water for injection extremely
Aequum, adjusts pH value 6.5~8.0, and it is filling then to rush nitrogen, sealing, with 100 DEG C of flowing steam sterilization 30 minutes, obtains final product
LEMAHUI ZHUSHEYE.
5. the preparation method of the LEMAHUI ZHUSHEYE of diabetes is treated as claimed in claim 4, it is characterised in that described to control
The preparation method of the LEMAHUI ZHUSHEYE of diabetes is treated, step is as follows:
(1)It is in 75% ethanol water, to be warming up to 85 DEG C to take dry 5 times of amount concentration of addition of reining in the horse back, is heated to reflux 1.5
Cold filtration after h, obtains extract and the dregs of a decoction, and to the dregs of a decoction aforesaid operations 2 times are repeated, and merges 3 gained liquids, and then decompression is steamed
Evaporate, obtain alcohol extract;
(2)Distilled water is added in alcohol extract, after being well mixed, after standing 4 h, is filtered, take supernatant, then use milipore filter mistake
Filter, filtrate concentration, it is extract at the beginning of 1.15 rein in the horse back to obtain relative density;
(3)Extract Jing D101B types macroporous absorbent resin at the beginning of reining in the horse back is processed, first with the water that the pH of 5 times of amounts of column volume is 3
Solution is eluted, and collects eluent, then is eluted with 50% ethanol of 5 times of amounts of column volume, collects eluent, is eluted twice
Liquid merges, concentration, obtains concentrate, then concentrate is adjusted into pH to 10 with ammoniacal liquor, and 1.5 h are heated at 60 DEG C, then uses salt
Then acid for adjusting pH is processed its Jing polyamide resin column to 5, with 3 times of water elution, eluent is collected, to washing at 80 DEG C
De- liquid is distilled, and removes ethanol extremely without alcohol taste, then adjusts pH to 7, obtains extract of reining in the horse back;
(4)Add sodium chloride and other drugs auxiliary material to be uniformly mixed in extract of reining in the horse back, be supplemented with water for injection extremely
Aequum, adjusts pH value 7, and it is filling then to rush nitrogen, sealing, with 100 DEG C of flowing steam sterilization 30 minutes, obtains final product re-injection of reining in the horse
Penetrate liquid.
6. the preparation method of the LEMAHUI ZHUSHEYE of diabetes is treated as claimed in claim 5, it is characterised in that step
(1)Described reduced pressure distillation process condition be -0.05~-0.09MPa, 40~90 DEG C.
7. the preparation method of the LEMAHUI ZHUSHEYE of diabetes is treated as claimed in claim 6, it is characterised in that step
(2)Milipore filter used by described ultrafiltration is 3000~6000 dalton, selected from cellulose diacetate film, Triafol T
Film, cyanoethyl cellulose film, PS membrane, sulfonated polysulfone membrane, poly (ether sulfone) film, sulfonated polyether sulfone film, Polysulfonamide, phenolphthalein
Side base polyarylsulfone (PAS) film, polyvinylidene fluoride film, polyacrylonitrile film, polyimide film, cellulose membrane, methyl methacrylate-propylene
Lonitrile copolymer film, polyacrylonitrile-cellulose diacetate blend film.
8. application of a kind of LEMAHUI ZHUSHEYE in treatment diabetes medicine is prepared, it is characterised in that the re-injection of reining in the horse is penetrated
Liquid is made up of active medicine and excipient substance, and the active medicine is extract of reining in the horse back, accounts for LEMAHUI ZHUSHEYE cumulative volume
5%, the excipient substance is sodium chloride, glycerine, glycerine, water, and the preparation method of the LEMAHUI ZHUSHEYE, step is as follows:
(1)It is in 75% ethanol water, to be warming up to 85 DEG C by dry 5 times of amount concentration of addition of reining in the horse back, is heated to reflux 1.5
Cold filtration after h, obtains extract and the dregs of a decoction, and to the dregs of a decoction aforesaid operations 2 times are repeated, and merges 3 gained liquids, then technique bar
Part is -0.05~-0.09MPa, vacuum distillation under conditions of 40~90 DEG C, obtains alcohol extract;
(2)Distilled water is added in alcohol extract, after being well mixed, after standing 4 h, is filtered, take supernatant, then with 3000~6000
The cellulose diacetate ultrafiltration membrance filter of dalton, filtrate concentration, it is extract at the beginning of 1.15 rein in the horse back to obtain relative density;
(3)Extract Jing D101B types macroporous absorbent resin at the beginning of reining in the horse back is processed, first with the water that the pH of 5 times of amounts of column volume is 3
Solution is eluted, and collects eluent, then is eluted with 50% ethanol of 5 times of amounts of column volume, collects eluent, is eluted twice
Liquid merges, concentration, obtains concentrate, then concentrate is adjusted into pH to 10 with ammoniacal liquor, and 1.5 h are heated at 60 DEG C, then uses salt
Then acid for adjusting pH is processed its Jing polyamide resin column to 5, with 3 times of water elution, eluent is collected, to washing at 80 DEG C
De- liquid is distilled, and removes ethanol extremely without alcohol taste, then adjusts pH to 7, obtains extract of reining in the horse back;
(4)Add sodium chloride and other drugs auxiliary material to be uniformly mixed in extract of reining in the horse back, be supplemented with water for injection extremely
Aequum, adjusts pH value 7, and it is filling then to rush nitrogen, sealing, with 100 DEG C of flowing steam sterilization 30 minutes, obtains final product re-injection of reining in the horse
Penetrate liquid.
9. application of a kind of LEMAHUI ZHUSHEYE in medicament for treating insomnia is prepared, it is characterised in that the LEMAHUI ZHUSHEYE by
Active medicine and excipient substance are constituted, and the active medicine is extract of reining in the horse back, accounts for the 5% of LEMAHUI ZHUSHEYE cumulative volume, institute
It is sodium chloride, glycerine, glycerine, water to state excipient substance, and the preparation method of the LEMAHUI ZHUSHEYE, step is as follows:
(1)It is in 75% ethanol water, to be warming up to 85 DEG C by dry 5 times of amount concentration of addition of reining in the horse back, is heated to reflux 1.5
Cold filtration after h, obtains extract and the dregs of a decoction, and to the dregs of a decoction aforesaid operations 2 times are repeated, and merges 3 gained liquids, then technique bar
Part is -0.05~-0.09MPa, vacuum distillation under conditions of 40~90 DEG C, obtains alcohol extract;
(2)Distilled water is added in alcohol extract, after being well mixed, after standing 4 h, is filtered, take supernatant, then with 3000~6000
The cellulose diacetate ultrafiltration membrance filter of dalton, filtrate concentration, it is extract at the beginning of 1.15 rein in the horse back to obtain relative density;
(3)Extract Jing D101B types macroporous absorbent resin at the beginning of reining in the horse back is processed, first with the water that the pH of 5 times of amounts of column volume is 3
Solution is eluted, and collects eluent, then is eluted with 50% ethanol of 5 times of amounts of column volume, collects eluent, is eluted twice
Liquid merges, concentration, obtains concentrate, then concentrate is adjusted into pH to 10 with ammoniacal liquor, and 1.5 h are heated at 60 DEG C, then uses salt
Then acid for adjusting pH is processed its Jing polyamide resin column to 5, with 3 times of water elution, eluent is collected, to washing at 80 DEG C
De- liquid is distilled, and removes ethanol extremely without alcohol taste, then adjusts pH to 7, obtains extract of reining in the horse back;
(4)Add sodium chloride and other drugs auxiliary material to be uniformly mixed in extract of reining in the horse back, be supplemented with water for injection extremely
Aequum, adjusts pH value 7, and it is filling then to rush nitrogen, sealing, with 100 DEG C of flowing steam sterilization 30 minutes, obtains final product re-injection of reining in the horse
Penetrate liquid.
10. a kind of application of LEMAHUI ZHUSHEYE in treatment medicine for treating diabetic nephropathy is prepared, it is characterised in that the re-injection of reining in the horse
Penetrate liquid to be made up of active medicine and excipient substance, the active medicine is extract of reining in the horse back, accounts for LEMAHUI ZHUSHEYE cumulative volume
5%, the excipient substance is sodium chloride, glycerine, glycerine, water, and the preparation method of the LEMAHUI ZHUSHEYE, step is as follows:
(1)It is in 75% ethanol water, to be warming up to 85 DEG C by dry 5 times of amount concentration of addition of reining in the horse back, is heated to reflux 1.5
Cold filtration after h, obtains extract and the dregs of a decoction, and to the dregs of a decoction aforesaid operations 2 times are repeated, and merges 3 gained liquids, then technique bar
Part is -0.05~-0.09MPa, vacuum distillation under conditions of 40~90 DEG C, obtains alcohol extract;
(2)Distilled water is added in alcohol extract, after being well mixed, after standing 4 h, is filtered, take supernatant, then with 3000~6000
The cellulose diacetate ultrafiltration membrance filter of dalton, filtrate concentration, it is extract at the beginning of 1.15 rein in the horse back to obtain relative density;
(3)Extract Jing D101B types macroporous absorbent resin at the beginning of reining in the horse back is processed, first with the water that the pH of 5 times of amounts of column volume is 3
Solution is eluted, and collects eluent, then is eluted with 50% ethanol of 5 times of amounts of column volume, collects eluent, is eluted twice
Liquid merges, concentration, obtains concentrate, then concentrate is adjusted into pH to 10 with ammoniacal liquor, and 1.5 h are heated at 60 DEG C, then uses salt
Then acid for adjusting pH is processed its Jing polyamide resin column to 5, with 3 times of water elution, eluent is collected, to washing at 80 DEG C
De- liquid is distilled, and removes ethanol extremely without alcohol taste, then adjusts pH to 7, obtains extract of reining in the horse back;
(4)Add sodium chloride and other drugs auxiliary material to be uniformly mixed in extract of reining in the horse back, be supplemented with water for injection extremely
Aequum, adjusts pH value 7, and it is filling then to rush nitrogen, sealing, with 100 DEG C of flowing steam sterilization 30 minutes, obtains final product re-injection of reining in the horse
Penetrate liquid.
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CN109793847A (en) * | 2019-04-09 | 2019-05-24 | 伍治明 | A kind of composite isatis root acupoint injection therapy liquid and preparation method thereof and application method |
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CN1634413A (en) * | 2004-10-29 | 2005-07-06 | 张平 | Bastard speedwell injection and preparation method thereof |
CN101564461A (en) * | 2009-06-12 | 2009-10-28 | 陕西东泰制药有限公司 | Method for preparing Chinese medicine used for clearing heat, moistening lung, relieving cough, resolving phlegm, disinhibiting urine and freeing strangury |
CN104383104A (en) * | 2014-10-23 | 2015-03-04 | 王�锋 | Application of bastard speedwell injection for preparing rectal administration preparation and aerosol inhalation preparation |
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2016
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CN1634413A (en) * | 2004-10-29 | 2005-07-06 | 张平 | Bastard speedwell injection and preparation method thereof |
CN101564461A (en) * | 2009-06-12 | 2009-10-28 | 陕西东泰制药有限公司 | Method for preparing Chinese medicine used for clearing heat, moistening lung, relieving cough, resolving phlegm, disinhibiting urine and freeing strangury |
CN104383104A (en) * | 2014-10-23 | 2015-03-04 | 王�锋 | Application of bastard speedwell injection for preparing rectal administration preparation and aerosol inhalation preparation |
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CN109793847A (en) * | 2019-04-09 | 2019-05-24 | 伍治明 | A kind of composite isatis root acupoint injection therapy liquid and preparation method thereof and application method |
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Effective date of registration: 20180207 Address after: 150040 Heping Road, Xiangfang District, Heilongjiang, Harbin, China, 24 Patentee after: Heilongjiang University of Chinese Medicine Address before: Oblitera.ns 150009 in Heilongjiang province Harbin city Nangang District guogeli Street No. 411 the Second Affiliated Hospital of Heilongjiang University Of Chinese Medicine Patentee before: Wang Jing |