CN1546091A - Use of ginkgo leaf extract - Google Patents
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- CN1546091A CN1546091A CNA2003101094455A CN200310109445A CN1546091A CN 1546091 A CN1546091 A CN 1546091A CN A2003101094455 A CNA2003101094455 A CN A2003101094455A CN 200310109445 A CN200310109445 A CN 200310109445A CN 1546091 A CN1546091 A CN 1546091A
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Abstract
The invention discloses a novel use of ginkgo leaf extract, wherein the ginkgo leaf extract and findings are made into preparation through the conventional processes for protecting optical nerve, thus can be used as an ideal medicament for protecting ganglionic cells.
Description
Technical field:
The present invention relates to a kind of new purposes of Folium Ginkgo extract, particularly to the purposes of optic nerve protection.
Technical background:
Optic nerve protection is also not sustain damage or only be subjected to part damage at those, itself is in the method that the neuron among the toxicity environment is protected.If these cells are not intervened, must be subjected to the murder by poisoning of environment and wither away gradually.For the unusual intensive disease of damage, the neuroprotective measure almost has little time to play a role.Therefore, neuroprotective is not suitable for the disease of the rapid disintegrate of cell, but is suitable for the disease that cell is slowly lost, as Alzhiemer disease and glaucoma etc.In nervous system lesion, primary injury (Primary injury) may be varied, comprise ischemia, anoxia, wound, degeneration, ion concentration change, oxygen-derived free radicals, neurotrophic factor shortage, dysimmunity etc., yet, neuronic Secondary cases is lost not because of the difference of primary affection and is changed to some extent, and they have all taked apoptotic mode.So, a kind of as nerve protection medicine, the optic nerve protection medicine is not at the specific cause of disease, but at common damage path basically, i.e. the retinal ganglion apoptosis that secondary lesion causes, and the various factors that causes apoptosis.
Glaucoma is a kind of main blinding oculopathy, and the visual function that is caused infringement can not reverse.According to World Health Organization's investigation, there are glaucoma patient more than 6,600 ten thousand people in the whole world, and crowd's sickness rate is 2% more than 40 years old, calculates according to this ratio, and along with the aggravation of aged tendency of population, glaucoma patient increases gradually.This disease has a strong impact on the health level that China influences the people, hinders the development of the economy.
Thought in the past that glaucoma was the disease that a kind of intraocular pressure increases, the intraocular pressure lowering medicine and the therapeutic method of surgery of a multitude of names that grows up clinically in order to reduce patient's intraocular pressure, are made every effort to the development of blocking condition.Up to the present, the intraocular pressure lowering treatment is the unique Therapeutic Method of glaucoma.Yet, in glaucoma patient,, patient's optic neuropathy continuation progress of 40% is arranged still even intraocular pressure is reduced to normal range, show as the further infringement in the visual field clinically, the further decline of visual function finally causes patient's blindness or blind.Even this is that a kind of neurocyte on optical fundus---the ganglion cell also can slowly lose because glaucoma was done operation or controlled intraocular pressure with additive method, this cell loss many more, visual function is just poor more, and is final blind.Therefore, people have the glaucoma of what is said or talked about greatly and the sensation of complexion changed.How protecting the ganglion cell is a current important topic.
Find that after deliberation glaucoma is a kind of optic neuropathy of chronic progressive external, it is its topmost pathophysiology feature that the chronic progressive external of retinal ganglion cell is lost.Primary injuryes (Primary injury) such as high intraocular pressure, ischemia at first cause losing of a part of retinal ganglion cell, the ganglion cell who loses produces many toxicants, make its living environment deterioration of ganglion cell on every side, the cell membrane ion permeability increases, the mitochondrial membrane sustain damage, cause cell to enter apoptosis program, cause cell loss, such damage is called as secondary lesion (Secondarydegeneration).Secondary lesion is sustained, and pathological changes is just made progress always.This shows that secondary lesion is the most important reason of glaucoma patient optic neuropathy progress.
There are two kinds of chemical synthetic drugs in the treatment glaucoma U.S. at present, memantine and Mo Niding, and FDA agrees to do the clinical experiment of medicine, does not also draw last result.How tame the synthetics of countries in the world laboratory research be a lot, finally all can not enter the clinical experiment result too greatly because of side effect.
Folium Ginkgo has had history in several thousand in the practicality of middle medical circles, it is found that many new purposes nearly decades.It contains flavone and lactone composition, effect with blood vessel dilating, antioxidation-anti-platelet aggregation, anti-nervous tissue, be specially adapted to the disease of neuronal apoptosis such as parkinsonism, extra large Mo's disease, obtain good result, this medicine is subjected to extensive welcome in American-European countries.Particularly, use very extensive in Germany.Also there is the application Folium Ginkgo extract to treat at present in the ophthalmic diseases field; but mainly concentrate on retinal arteriosclerosis, pathological changes, improve the circulation of retina blood; and the treatment of aspect such as asthenopia, myopia, for the also more not deep research in the protection aspect of optic nerve.Domestic have that a kind of medicine---Herba Erigerontis (being the positive control medicine of selecting for use in the zoopery of the present invention) has certain effect to the protection of optic nerve, but should the non-medical insurance scope of product genus, and price is higher.
By experiment in recent years, think that tentatively Folium Ginkgo extract has possessed the condition of optic nerve protection medicine, at first, can confirm that by zoopery it has protective effect to the retinal ganglion cell; The second, from present research, what play a major role in numerous compositions of Folium Ginkgo extract has two classes, and ginkgetin and lactone composition are brought into play the effect of antioxidation and blood vessel dilating respectively.In addition, it can suppress the nmda receptor toxicity excitement that glutamic acid causes, suppress the cell injury that the paf receptor excitement causes, the precursor (APP) that suppresses amyloid transforms to amyloid, and these have all set forth the feasibility of Folium Ginkgo extract as the optic nerve protection agent from the molecular biology level; The 3rd, this medicine can reach target cell, and through giving the oral Folium Ginkgo extract of animal, finding has enough this medicine and metabolite thereof on retina, illustrate that Folium Ginkgo extract can pass through blood-eye barrier, reaches the retinal ganglion cell and brings into play protective effect.Studies show that in recent years, Folium Ginkgo extract has good action to the artificial diabetes retinopathy, and this reflects it from another side and can play a role at retina.
Summary of the invention:
The objective of the invention is to: a kind of optic nerve protection medicine made from Folium Ginkgo extract is provided, can sees through blood-eye barrier and directly act on retina, retinal blood flow is increased, and can directly expand optical fundus blood vessel; Also have anti-platelet aggregation, antagonism nmda receptor, antioxidation, antagonism apoptosis, on retina, reach effects such as effective drug level, to reach the purpose of diseases such as repairing and protect retinal ganglion cell and then treatment glaucoma.
The present invention is achieved in that it is to take common process to make preparation Folium Ginkgo extract and adjuvant, is used for the protection of optic nerve.
Described preparation can be capsule, tablet, injection, oral liquid, granule.
Described optic nerve protection comprises glaucomatous treatment.
The part by weight of Folium Ginkgo extract and adjuvant is in the described preparation: 20%-80%: 80%-20%.
Described optic nerve protection is to the reparation of retinal ganglion cell and protection.
The present invention compared with prior art because it comes from plant, contains more than 60 kind of composition, and wherein more than 30 kind of composition all is that it is peculiar at occurring in nature, and it is little to have toxic and side effects, at nervous system disease and cardiovascular disease unique advantages such as curative effect is arranged.
Below the present invention being crushed the model ganglion cell to the rat optic nerve protects the protection experiment that reaches the high intraocular pressure ganglion cell of rat acute to elaborate.
Description of drawings:
Accompanying drawing 1 crushes the contrast figure that the model ganglion cell protects survival rate for the present invention to the rat optic nerve;
Accompanying drawing 2 is protected medication amount effect relationship curve chart for the present invention crushes the model ganglion cell to the rat optic nerve;
Accompanying drawing 3 is protected the contrast figure of survival rate to the high intraocular pressure ganglion cell of rat optic nerve for the present invention;
Accompanying drawing 4 is protected medication amount effect relationship curve chart for the present invention to the high intraocular pressure ganglion cell of rat optic nerve.
One, optic nerve is crushed the experiment of model ganglion cell protective effect
(1), experiment purpose:
Crush model by optic nerve, observe ginkgo biloba p.e to pattern of retinal ganglion cells Protective effect, for clinical practice provides experimental basis.
(2), experiment material
Animal used as test
The SD rat is used in experiment, and the SPF level is identified institute's breeding field available from Chinese pharmaceutical biological product.
Experimental drug
Experimental drug ginkgo biloba p.e medicinal extract (hereinafter to be referred as Gin Kgo Plus) applicant provides
Positive control drug erigeron breviscapus medicinal extract is available from all places, Yuxi, Yunnan Province natural drug Co., Ltd
Negative control medicine normal saline North China pharmaceutcal corporation, Ltd
Drive in the wrong direction label fluorogold (FG) available from U.S. fluorogold company (Fluorochrome LtD)
Narcotics ketaject injection Shanghai No.1 Bio-Chemical Pharmacetical Industry Co., Ltd
Diazepam injection Tianjin gold credit aminoacid company limited
The fixative paraformaldehyde is available from Beijing chemical reagent product company limited
(3), experimental technique
1, animal and grouping: experiment is with 90 of SD rats, and is male, body weight 200-240 gram, and preocular inspection, the examination of ocular fundus of Mydrine mydriasis are no abnormal.With table of random number rat is divided into 5 groups at random: be respectively negative control group, positive controls, low dosage medication group, middle dosage medication group and high dose medication group, each 18 of every group of rats.Each organizes the rat right eye is harmed eye, and left eye is not done the optic nerve clamping, as normal control.
2, crush model and medication: rats by intraperitoneal injection ketalar 3ml/kg anaesthetizes, and cuts off outer canthus, and (OMS-85, topological Kanggong department Japan) cuts off bulbar conjunctiva and does corresponding separation under operating microscope, exposes optic nerve.With the miniature optic nerve folder (the TKF-2 type U.S.) of chucking power 40g 2mm place clamping optic nerve 60 seconds behind ball, the recovery bulbar conjunctiva is also sewed up the outer canthus wound.Visible one property crossed optical fundus blood vessel spasm crushes in back 2 minutes and recovers fully during the clamping optic nerve.Negative control group, positive controls, low dosage medication group, middle dosage medication group and high dose medication group crushed back 2 hours and respectively injecting normal saline 5ml/kg, 1% Herba Erigerontis 5ml/kg, 0.25% Gin Kgo Plus 5ml/kg, 1% Gin Kgo Plus 5ml/kg and 4% Gin Kgo Plus 5ml/kg of abdominal cavity every day after this in optic nerve, and experimental period is 28 days.
3, retrograde labelling: preceding 5 days of animal mercy killing, adopt the retrograde labelling retinal ganglial cells of two superior colliculus injections.6% chloral hydrate 0.5ml/100g lumbar injection makes Animal Anesthesia, be fixed in (the 900 type brain solid positioner David Kopf instrument company U.S.) on the brain solid positioner, dental burr is bored and is opened skull, draw 3% fluorogold (being dissolved in 0.9% normal saline) with microsyringe, every side superior colliculus is injected 2 points (behind the bregma-5.9 and-6.4, the side opens 1.4, and dark 4.0), every some injection 1.5ul.
4, retina shop sheet: bulbar conjunctiva is located to meet wire tag in 12 of tops, takes out eyeball immediately and is fixed in the 4% paraformaldehyde phosphate buffer 2 hours.0.5mm cuts off eyeball behind the limbus of corneae, removes cornea and crystalline lens, separates retina and with its directed tiling, natural drying in the air.Vectashield H-100 fluorescence-enhancing agent and commercially available colourless nial polish mounting.Fluorescence microscope is taken pictures (AH-2 Olympic Ba Si company Japan), and distance is looked nipple center 2mm and respectively taken pictures up and down one, photo amplification 125 *.
5, graphical analysis: use the Mustek of association scanner that photo is swept computer with 100dpi, use grading analysis software (CPAS) to carry out ganglion cell's counting, 4 photo ganglion cell quantity add up up and down, and change into ganglion cell's density.Right eye ganglion cell density is defined as ganglion cell's survival rate with the ratio of left eye ganglion cell density, i.e. ganglion cell's survival rate=right eye ganglion cell density/left eye ganglion cell density * 100.
6, statistical method: use SPSS for Windows 10.0 statistical packages, adopt one factor analysis of variance (ANOV), relatively adopt mean t check in twos.Have significant difference with p＜0.05, have highly significant difference with p＜0.01.
(4), experimental result
Because anesthesia and operation accident cause the part rats death, and indivedual rats can not use specimen because of ganglion cell's labelling is bad, the actual spendable rat quantity of each experimental group sees Table 1 in the experiment.Experimental result shows that negative control group ganglion cell's survival rate is between 41.27% and 74.21%, and average survival rate is 60.59%, and standard deviation is ± 8.78%.Calculate by ganglion cell's survival rate, positive controls, low dosage medication group, middle dosage medication group and high dose medication group ganglion cell's survival rate is respectively 73.80%, 66.49% and 71.60% and 74.17.Each is organized ganglion cell's survival rate and meansigma methods and standard deviation thereof and sees Table 2.Each organizes average ganglion cell's survival rate and standard deviation is seen Fig. 1.
Respectively organize final rat quantity (only) in table 1 experiment
Dosage group high dose group in the negative control positive control low dose group
Size of animal 18 16 15 18 16
Each one factor analysis of variance of organizing ganglion cell's survival rate shows, has significant difference (p＜0.05) between them.Relatively show in twos, have significant difference (p＜0.05 sees Table 3) between negative control group and positive controls, low dosage medication group, middle dosage medication group and the high dose medication group.The Gin Kgo Plus that positive control medicine Herba Erigerontis and three dosage groups are described can both be protected retinal ganglial cells effectively, each the group between relatively see Table 3 in twos.
Among the medication group, there is significant difference (p=0.032) between low dosage medication group and the negative control group, there is significant difference (p=0.038) between middle dosage medication group and the low dosage medication group, and unknown significance difference (p=0.33) between high dose medication group and the middle dosage medication group, but its ganglion cell's survival rate still increases, and brings up to 74.17% from 71.60%.The dose-effect relationship of three dosage groups is seen Fig. 2.As can be seen from the figure, the Gin Kgo Plus of low dosage just can be protected the ganglion cell, and along with the increase of dosage, its protective effect strengthens gradually, and ganglion cell's protective effect of Gin Kgo Plus has reasonable dose-effect relationship.
There is significant difference (p=0.023) in ganglion cell's survival rate between positive controls and the low dosage medication group, and both drug doses have also differed 4 times.When dosage is identical, in the promptly middle dosage group, just with positive controls there was no significant difference (p=0.48).This research is also found, there was no significant difference (p=0.92) between positive controls and the high dose medication group.
Table 2. is respectively organized ganglion cell's survival rate, meansigma methods and standard deviation
Dosage group high dose group in the negative control positive control low dose group
50.59 57.23 68.98 59.39 79.42
61.52 72.51 66.47 85.88 74.16
61.78 76.13 60.27 62.96 64.04
68.90 50.8 66.11 72.34 75.17
65.55 80.61 77.18 73.36 80.86
63.28 75.74 75.15 73.66 88.00
53.66 55.7 60.53 76.76 69.24
53.90 80.75 71.62 73.45 77.30
51.25 80.20 69.32 80.97 73.40
41.27 76.12 58.59 76.56 78.30
72.54 74.88 65.56 77.22 74.57
64.43 83.42 62.86 62.6 70.25
50.21 84.12 62.35 77.28 84.24
67.03 73.24 70.84 68.76 73.68
68.38 86.28 61.45 66.61 61.93
74.21 73.08 76.39 62.09
61.87 58.89
60.33 65.79
Meansigma methods 60.59 73.80 66.49 71.60 74.17
Standard deviation 8.78 10.48 5.62 7.57 8.39
Table 3. is respectively organized the comparing in twos of ganglion cell's survival rate (p value)
Dosage group high dose group in the positive control low dose group
Negative control 0.0004 0.032 0.0003 0.00003
Positive control 0.023 0.48 0.92
Low dose group 0.038 0.003
Middle dosage group 0.33
(5), discuss
1, optic nerve crushes the model optic nerve and crushes the research that model is usually used in the optic nerve protection medicine, and the common method that crushes has been adopted in this research, promptly does the bulbar conjunctiva otch in the temporo side, 2mm clamping optic nerve behind ball.This method does not need excessive tractive optic nerve, exposes to know processing ease, the optic nerve injury basically identical that is caused.We use the miniature optic nerve of 40g to crush optic nerve 60 seconds, cause the ganglion cell on average to lose 39.41% ± 8.78%, and losing of model ganglion cell is more stable.Only cause retinal vessel spasm in short-term when crushing, do not cause blood vessel forever inaccessible, retinal vessel all recovered in 2 minutes, did not have a rat to be excluded outside experiment because of the permanent ischemia of retina in the experiment.
2, the effect Herba Erigerontis of Herba Erigerontis can be protected retinal ganglial cells significantly; with negative control group significant difference (p＜0.05) is arranged relatively; this result is consistent with research in the past, and this has also illustrated the stability and the reliability of this model from another aspect.1% Herba Erigerontis group has been compared significant difference (p＜0.05) with 0.25% Gin Kgo Plus group, the effective dose that is likely Herba Erigerontis is higher than 4 times of 0.25% Gin Kgo Plus, and the difference of drug dose has caused the difference to ganglion cell's protective effect.When Herba Erigerontis and Gin Kgo Plus under Isodose, concentration was all 1% o'clock, both ganglion cell's protective effect there was no significant differences (p＞0.05).
3, the effect experimental result of Gin Kgo Plus shows that the Gin Kgo Plus of basic, normal, high dosage group all has ganglion cell's protective effect.Low dose group ganglion cell's survival rate has improved 5.9% than negative control group, and middle dosage group ganglion cell survival rate improves 5.11% than low dose group.The dosage group improves 2.57% in high dose group ganglion cell's survival rate ratio.The Gin Kgo Plus of various dose shows good dose-effect relationship, and along with the raising of drug level, ganglion cell's protective effect constantly strengthens, and also exists significant difference (p＜0.05) between middle dosage group and the low dose group.Though there was no significant difference between middle dosage group and the high dose group (p＞0.05), the dosage group still increases in high dose group ganglion cell's survival rate ratio.
4, the mechanism of the effect of Gin Kgo Plus at first Gin Kgo Plus can arrive retina, adopt the method for labelled with radioisotope, give the oral Gin Kgo Plus of rat, the result shows that Gin Kgo Plus and metabolite thereof can reach certain drug level on retina.Secondly, Gin Kgo Plus has been brought into play vasodilative effect, studies show that, color Doppler shows that the ophthalmic artery blood flow has increased by 24% behind normal volunteer oral's Gin Kgo Plus.Think that at present Gin Kgo Plus has multiple effect, comprise that suppress the excitement of nmda receptor toxicity, the blocked glutamic acid toxic action suppresses platelet aggregation, antagonism apoptosis etc. to anti peroxidation of lipid.It has protective effect to the neuronal damage that ischemia/anoxia, mechanical damage cause.Recently studies show that extensively depositing platelet activating factor (PAF) receptor on the retina, its exaltation has caused losing of ganglion cell, and optic nerve lesion is constantly increased the weight of.Gin Kgo Plus can suppress the excitement of paf receptor, thereby reaches protection ganglion cell purpose.
In a word, Gin Kgo Plus can protect the rat optic nerve to crush the retinal ganglial cells of model effectively, just shows significant protective effect when low dosage, along with the raising ganglion cell protective effect of dosage is more obvious, good dose-effect relationship is arranged.Gin Kgo Plus belongs to the plant amedica category, and toxic and side effects is little, and is safe in utilization.
Two, to the experiment of the high Intraocular Pressure Model ganglion cell's protective effect of rat acute
(1), experiment purpose:
By the high intraocular pressure retinal ischemia of rat acute re-perfusion model, it is right to observe Gin Kgo Plus
The protective effect of retinal ganglion cell is for clinical practice provides experimental basis.
(2), experiment material
Laboratory animal
The SD rat is used in experiment, and the SPF level is identified institute's breeding field available from Chinese pharmaceutical biological product.
Experimental drug
Experimental drug Gin Kgo Plus extractum applicant provides
Positive control drug Herba Erigerontis extractum is available from all places, Yuxi, Yunnan Province natural drug company limited
Negative control medicine normal saline North China pharmaceutcal corporation, Ltd
Drive in the wrong direction label fluorogold (FG) available from U.S. fluorogold company (Fluorochrome LtD)
Narcotics ketaject injection Shanghai No.1 Bio-Chemical Pharmacetical Industry Co., Ltd
Diazepam injection Tianjin gold credit aminoacid company limited
The fixative paraformaldehyde is available from Beijing chemical reagent product company limited
(3), experimental technique
Laboratory animal and grouping
Experiment is with 90 of SD rats, and is male, body weight 200-240 gram, and preocular inspection, the examination of ocular fundus of Mydrine mydriasis are no abnormal.With table of random number rat is divided into 5 groups at random: be respectively negative control group, positive controls, low dosage medication group, middle dosage medication group and high dose medication group, each 18 of every group of rats.Each organizes the rat right eye is harmed eye, and left eye is not made the acute high intraocular pressure model, as normal control.
Experimental technique
1, ischemia-reperfusion injury model and medication: rats by intraperitoneal injection ketalar 3ml/kg anaesthetizes, right eye Mydrin-P mydriatic mydriasis, behind the platycoria, (OMS-85 under operating microscope, topology Kanggong department Japan) No. 7 children scalps that will connect normal saline bottle tube for transfusion are needled into the rat anterior chamber, avoid damaging iris and crystal, saline bottle to experimental eye vertical dimension 150cm place, this height can form the intraocular pressure of 110mmHg (15.96Kpa) within the eye, the visible bulbar conjunctiva of naked eyes this moment is pale, the visible retinal ischemia of ophthalmofundoscope was kept 60 minutes.Then reduce saline bottle and reach the animal level, transfer to syringe needle, visible conjunctival congestion, retina blood be perfusion more again.Stomach normal saline 5ml/kg, 1% Herba Erigerontis 5ml/kg (available from Yunnan Province), 0.25% Gin Kgo Plus 5ml/kg, 1% Gin Kgo Plus 5ml/kg and 4% Gin Kgo Plus 5ml/kg were damaged back 2 hours and after this irritated every day to negative control group, positive controls, low dosage medication group, middle dosage medication group and high dose medication group in acute high intraocular pressure.Experimental period is 28 days.
2, retrograde labelling: preceding 5 days of animal mercy killing, adopt the retrograde labelling retinal ganglial cells of two superior colliculus injections.6% chloral hydrate 0.5ml/100g lumbar injection makes Animal Anesthesia, be fixed in (the 900 type brain solid positioner David Kopf instrument company U.S.) on the brain solid positioner, dental burr is bored and is opened skull, draw 3% fluorogold (being dissolved in 0.9% normal saline) with microsyringe, every side superior colliculus is injected 2 points (behind the bregma-5.9 and-6.4, the side opens 1.4, and dark 4.0), every some injection 1.5ul.
3, retina shop sheet: bulbar conjunctiva is with the carbon element labelling up, takes out eyeball immediately and is fixed in the 4% paraformaldehyde phosphate buffer 2 hours.0.5mm cuts off eyeball behind the limbus of corneae, removes cornea and crystalline lens, separates retina and with its directed tiling, natural drying in the air.Vectashield H-100 fluorescence-enhancing agent and commercially available colourless nial polish mounting.Fluorescence microscope is taken pictures (AH-2 Olympic Ba Si company Japan), and distance is looked nipple center 2mm and respectively taken pictures up and down one, photo amplification 125 *.
The rejecting standard
1) paracentesis of anterior chamber causes intraocular hemorrhage and endophthalmitis, major injury iris and crystalline lens.2) the RGCs fluorescent labeling is bad, and the subregion labelling is not gone up.
Graphical analysis and statistical method
Use the Mustek of association scanner that photo is swept computer with 100dpi, use grading analysis software (CPAS) to carry out ganglion cell's counting, 4 photo ganglion cell quantity add up up and down, and change into ganglion cell's density.Right eye ganglion cell density is defined as ganglion cell's survival rate with the ratio of left eye ganglion cell density, i.e. ganglion cell's survival rate=right eye ganglion cell density/left eye ganglion cell density * 100.Use SPSS for Windows 10.0 statistical packages, adopt one factor analysis of variance (ANOV), relatively adopt mean t check in twos.Have significant difference with p＜0.05, have highly significant difference with p＜0.01.
(4), experimental result
Because anesthesia and operation accident cause the part rats death, indivedual rats can not use specimen because of ganglion cell's labelling is bad in the experiment.The actual spendable rat quantity of each experimental group sees Table 4.Negative control group ganglion cell's survival rate is between 53.97% and 76.59%, and average survival rate is 67.54%, and standard deviation is ± 7.50%.Calculate by ganglion cell's survival rate, positive controls, low dosage medication group, middle dosage medication group and high dose medication group ganglion cell's survival rate is respectively 74.25%, 73.47%, 78.85% and 81.14%.Each organizes ganglion cell's survival rate and meansigma methods sees Table 5.Each organizes average ganglion cell's survival rate and standard deviation is seen Fig. 3.
Respectively organize final rat quantity (only) in table 4 experiment
Dosage group high dose group in the negative control positive control low dose group
Size of animal 15 13 12 13 14
Each one factor analysis of variance of organizing ganglion cell's survival rate shows, has significant difference (p＜0.05) between them.Relatively show in twos, have significant difference (p＜0.05all groups) between negative control group and positive controls, low dosage medication group, middle dosage medication group and the high dose medication group.The Folium Ginkgo extract that positive control medicine Herba Erigerontis and three dosage groups are described can both be protected retinal ganglial cells effectively.The protective effect of high dose Folium Ginkgo extract is better than 1% Herba Erigerontis group.Each the group between relatively see Table 6 in twos.
Among the medication group, there is significant difference (p=0.0040) between low dosage medication group and the negative control group, there is significant difference (p=0.0178) between high dose medication group and the low dosage medication group, and unknown significance difference (p=0.0540) between high dose medication group and the middle dosage medication group, but its ganglion cell's survival rate still increases, and brings up to 81% from 79%.The dose-effect relationship of three dosage groups is seen Fig. 4.As can be seen from the figure, Gin Kgo Plus extractum low dosage just can be protected the ganglion cell, and along with the increase of dosage, its protective effect strengthens.Ganglion cell's protective effect of Gin Kgo Plus extractum has reasonable dose-effect relationship.
There is significant difference between positive controls and the high dose medication group, there is no significant difference between positive controls and the low middle dosage medication group.The drug level of the drug level of positive controls and middle dosage medication group is suitable, and is higher 4 times than low dosage medication group drug level.
Table 5. is respectively organized ganglion cell's survival rate (%)
Dosage group high dose group in the negative control positive control low dose group
74.86 82.10 67.75 77.96 71.86
67.16 62.91 64.14 78.07 85.29
67.82 70.53 73.52 90.42 79.00
66.42 64.50 78.69 65.43 82.84
82.26 71.15 73.84 74.68 87.69
68.45 83.48 75.32 76.33 84.91
60.12 77.14 82.47 82.30 77.48
59.23 66.00 73.78 73.08 78.71
66.60 80.81 71.83 83.60 68.38
53.97 77.46 75.99 80.49 82.94
71.34 74.17 76.43 80.57 82.23
74.39 76.96 67.94 83.07 87.52
61.28 78.04 79.11 81.57
62.67 85.57
76.59
Meansigma methods 67.54 74.25 73.47 78.85 81.14
Standard deviation 7.50 6.74 5.05 5.99 5.65
Table 6. respectively organize ganglion cell's survival rate comparison (p value)
Dosage group high dose group in the positive control low dose group
Negative control 0.02 0.03 0.0002 0.00008
Positive control 0.75 0.08 0.008
Low dose group 0.02 0.001
Middle dosage group 0.32
(5), discuss
1, about the acute high intraocular pressure model: research ganglion cell's injury characteristic and optic nerve protection medicine, utilize the high Intraocular Pressure Model model of rat acute comparative maturity.In our experiment, adopt the method rising intraocular pressure of paracentesis of anterior chamber, and keep 110mmHg, continue 60 minutes.This pressure makes the complete ischemia of rat retina, and this point can be proved by observing optical fundus blood vessel.The advantage of this animal model is the term harmonization of experimental eye, and as identical pressure and identical persistent period, simple to operate, easy to control, expense is low, and this protection research and screening nerve protection medicine for further retinal neuronal cell is laid a good foundation.We selected ischemia 60 minutes in this experiment, and the average survival rate of ganglion cell is 67.54 ± 7.50% in the negative control group, and it is more stable that this model ganglion cell loses ratio.After Selles-Navarro I etc. discovered and uses the retrograde labelling RGCs of the capable superior colliculus of fluorogold, the existence of fluorogold was no more than for 3 weeks in the endochylema.Progress may fluorogold be lost fluorescence or by metabolism in time.This is studied in preceding 5 days retrograde labelling superior colliculus of capable fluorogold of sacrifice of animal, and the RGCs of labelling is golden yellow hyperfluorescence, and all quadrants is even.By the retrograde labelling RGCs of fluorogold superior colliculus, be convenient to quantitative, the qualitative investigation of RGCs.
2, ganglion cell's protective effect of Herba Erigerontis: our result is consistent with research in the past; with ganglion cell's survival rate is index; Herba Erigerontis has the effect of significant protection retinal ganglial cells, with negative control group significant difference (P＜0.05) is arranged relatively.Ganglion cell's protective effect of 1% Herba Erigerontis group and 4% Gin Kgo Plus group have significant difference (P＜0.05); and do not have significant difference (P＞0.05) with 0.25% and 1% Gin Kgo Plus group; show that 1% Herba Erigerontis and 0.25% and 1% Gin Kgo Plus neuroprotective ganglion cell act on quite, 4% Gin Kgo Plus is stronger than ganglion cell's protective effect of 1% Herba Erigerontis.
3, ganglion cell's protective effect of Gin Kgo Plus: we are index by the high Intraocular Pressure Model of rat acute with ganglion cell's survival rate, and the result shows that the Gin Kgo Plus of basic, normal, high dosage group all has ganglion cell's protective effect.Low dose group ganglion cell's survival rate has improved 5.93% than negative control group, and middle dosage group has improved 5.38% than low dose group, and the dosage group has improved 2.29% in the high dose group ratio.The Gin Kgo Plus of various dose shows good dose-effect relationship.Along with the raising of drug level, ganglion cell's protective effect constantly strengthens, though there was no significant difference between middle dosage group and the high dose group, high dose group ganglion cell's survival rate increases.
4, the mechanism of Gin Kgo Plus ganglion cell protective effect: ginkgo biloba succi mainly contains 24% flavonol glycosides, 6% lactone and 7% anthocyanidin precursor, also has some not qualitative chemical compounds.The mechanism of its protection optic nerve may be: 1. the effect that has the antagonism platelet activating factor; the Semen Ginkgo lactone is considered to have most the natural platelet activating factor receptor antagonist of potential applicability in clinical practice at present; 2. removing free radical; reproducibility hydroxyl functional group in the Folium Ginkgo extract; can directly bring into play antioxidation; remove oxygen-derived free radicals, alleviate or block retinal function and morphologic damage that lipid generation peroxidating causes.3. to sanguimotor influence, discover that ginkgo extract can increase the blood flow of cerebrovascular and peripheral vessels, make the ophthalmic artery blood flow increase by 24%.4. to the protective effect of ischemic injuries, find that the Semen Ginkgo lactone can improve the infringement of ischemia reperfusion injury to myocardium regulating liver-QI, improve the hepatic ischemia Rats survival rate, significantly reduce the hepatic ischemia damage.Simultaneously, it can also reduce the overload of the intracellular calcium of glutamate induction.
In a word, Gin Kgo Plus and oil lamp carefulness can both protect the rat optic nerve to crush the retinal ganglial cells of model, and Gin Kgo Plus is a Folium Ginkgo extract, belongs to the plant amedica category, and toxic and side effects is little, and is safe in utilization.Its effect is likely many sides, many target spots.
The specific embodiment:
Embodiments of the invention: with the capsule of producing 40 milligrams of specifications of Folium Ginkgo extractum is example:
With reference to Ministry of Health of the People's Republic of China's ministry standard
Prescription: in 100,000 unit: kg
Folium Ginkgo dry extract 4.00
Lactose 14.82
Magnesium stearate 0.29
Micropowder silica gel 0.39
Technology: get Folium Ginkgo extractum and add lactose and mix and to beat powder, granulate, mixing with it after magnesium stearate, micropowder silica gel are sieved, granulate is encapsulated then.Every contains Folium Ginkgo extractum 40mg, contains total flavonoids 9.6mg.
In the zoopery result aspect the glaucoma ganglion cell protection, suggestion adult taking dose is 1～2/day according to Folium Ginkgo extract.
The also available common process of the present invention is made injection, tablet, oral liquid, granule.
Claims (5)
1, a kind of novel use of ginkgo leaf extract is characterized in that: it is to take common process to make preparation Folium Ginkgo extract and adjuvant, is used for the protection of optic nerve.
2, according to the described novel use of ginkgo leaf extract of claim 1, it is characterized in that: described preparation can be capsule, tablet, injection, oral liquid, granule.
3, according to the described novel use of ginkgo leaf extract of claim 1, it is characterized in that: described optic nerve protection comprises glaucomatous treatment.
4, according to claim 1 or 2 described novel use of ginkgo leaf extract, it is characterized in that: the part by weight of Folium Ginkgo extract and adjuvant is in the described preparation: 20%-80%: 80%-20%.
5, according to claim 1 or 3 described novel use of ginkgo leaf extract, it is characterized in that: described optic nerve protection is to the reparation of retinal ganglion cell and protection.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102078343A (en) * | 2010-12-21 | 2011-06-01 | 上海信谊百路达药业有限公司 | Application of ginkgo leaf total lactones in preparation of medicament for preventing or treating deafness and tinnitus |
| CN104398543A (en) * | 2014-12-05 | 2015-03-11 | 上海信谊百路达药业有限公司 | Application of folium ginkgo composition to preparation of medicine for preventing or treating glaucoma |
| CN110678171A (en) * | 2017-05-15 | 2020-01-10 | 坪田实验室股份有限公司 | Composition and functional food for preventing myopia |
-
2003
- 2003-12-16 CN CN 200310109445 patent/CN1221279C/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102078343A (en) * | 2010-12-21 | 2011-06-01 | 上海信谊百路达药业有限公司 | Application of ginkgo leaf total lactones in preparation of medicament for preventing or treating deafness and tinnitus |
| CN102078343B (en) * | 2010-12-21 | 2012-10-03 | 上海信谊百路达药业有限公司 | Application of ginkgo leaf total lactones in preparation of medicament for preventing or treating deafness and tinnitus |
| CN104398543A (en) * | 2014-12-05 | 2015-03-11 | 上海信谊百路达药业有限公司 | Application of folium ginkgo composition to preparation of medicine for preventing or treating glaucoma |
| CN110678171A (en) * | 2017-05-15 | 2020-01-10 | 坪田实验室股份有限公司 | Composition and functional food for preventing myopia |
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