| First part:A literature study on the treatment of optic atrophy of Chinese MedicineObjective:to analyze the clinical rule by analyzing the modern literature of the treatment of optic atrophy by traditional Chinese medicine,so as to provide evidence for clinical differentiation and treatment of optic atrophy.Methods:using the computer retrieval:Chinese journal full text database(CNKI),Chinese Journal Database(VIP),Wanfang database knowledge service platform(WanFangData)on optic atrophy of traditional Chinese medicine treatment related literature,using Note express for various database retrieval of literature collection and screening,using Micro Excel in soft office records the essential elements of collecting literature,establishes corresponding database,and applies SPSS 19.0 to analyze and process corresponding data.Results:(1)descriptive statistics showed that the highest frequency of drug use was Angelica(81.73%),indicating that on the clinical treatment of blindness,the treatment from the blood and the liver is a more common method.According to the classification and analysis of the efficacy and treatment,33.32%of the drugs used were used,and 16.66%of the drugs used for activating blood and removing stasis were used.It showed that the disease was mostly treated from Qi and blood,deficiency of Qi and blood or qi stagnation and blood stasis,Qi deficiency and blood stasis,etc.(2)Cluster analysis found that the frequency of Chinese herbs with frequency over 50 times is 16,and it can be synthesized into the following 3 categories.Among them,the main drugs in cluster 1 include Poria cocos,Atractylodes macrocephala,licorice root,Radix Paeoniae Alba and Angelica sinensis,and their main functions are tonifying qi and generating blood.This class of drugs derived from Bazhen decoction.The main drugs included in the cluster 2 include Radix Bupleuri,calamus,Ligusticum chuanxiong,safflower,raw land,Radix Paeoniae,radix salviae miltiorrhizae,and Astragalus membranaceus.The main effect is qi activating blood circulation,and these drugs are derived from the Xuefu Zhuyu decoction or the Five Decoction of tonifying yang and returning Yang.The main drugs in cluster 3 include wolfberry fruit,Chinese yam and Rehmannia glutinosa,and their main functions are tonifying liver and kidney,which are derived from YouGui pill or Zuo Gui pills.(3)through factor analysis,it was found that using the 15 traditional Chinese medicine and using 6 public factors,the percentage of the cumulative variance was 62.969%,more than 0.5,and the factor analysis results were acceptable.The number of loads is greater than 0.5,which means that the change is more important than the factor.According to this principle,the importance of the 15 flavors is classified.Altogether divided into 6 categories,namely,the public factor 1 categories include angelica,Poria cocos,licorice,Atractylodes,Paeonia lactiflora,2 types of public factors include safflower and Ligusticum chuanxiong;3 of public factors include the land and red peony;the 4 public factors include Lycium barbarum and cooked land;the 5 of the public factors include Shi Changpu and Radix Bupleuri,and the 6 public factors include Salvia miltiorrhiza.Astragalus,Astragalus.According to the theory of traditional Chinese medicine and consulting experts,the names of public factors are determined.The final 6 categories are named"replenishing qi and nourishing blood,regulating spleen and stomach","nourishing liver and kidney","clearing heat and nourishing Yin","activating blood and removing blood stasis","activating blood and removing stasis" and "doing blood camp"Conclusion:By collecting and analyzing the literature study on the treatment of optic atrophy published by Chinese traditional medicine,it is found that the treatment of optic atrophy in Chinese medicine is mainly guided by the theory of liver and the theory of Qi and blood.At the same time,the common syndromes and prescriptions in the clinic are Qi and blood deficiency,deficiency of liver and kidney,qi stagnation and blood stasis and eight treasure soup,Zuo GUI pill or right return pill and Xuefu Zhuyu Decoction.The commonly used changes are mainly from 6 aspects,such as "Nourishing Qi and nourishing blood,regulating spleen and stomach","nourishing liver and kidney","clearing heat and nourishing Yin","activating blood and removing blood stasis","activating blood and removing stasis" and "running blood camp" and so on.Second part:Experimental study on the protection of optic nerve of Astragalus PolysaccharideObjective:To make the rat model of optic nerve injury by lateral quantitative traction,and to observe the effect of Astragalus Polysaccharides on the PI3K/AKT signal pathway of retina tissue from the angle of histology and molecular biology,and to explore the possible molecular mechanism of the protection of Astragalus Polysaccharide to optic nerve.Methods:first,the lateral quantitative traction method was used to construct the animal model of optic nerve injury,and the retrograde labeling of retinal ganglion cells(RGCs)was used to detect the survival rate of RGCs,and the feasibility of making the animal model of optic nerve injury by lateral quantitative traction was evaluated.After the model was successful made,the rats were randomly selected.All rats were radomely divided into 6 groups:blank group,sham operation group,model group,low,medium and high dose group.The sham operation group only exposed the optic nerve,the blank group,the sham operation group and the model group were given the saline for 14 days by intraperitoneal injection of saline,and the APS treatment group was treated with different doses respectively.After 14 days of treatment,Western Blot and Real-time PCR were used to determine the expression of related indexes on the PI3K/AKT signaling pathway in retinal tissue.Results:(1)the results of animal model evaluation:the model of optic nerve injury in rats was made by lateral quantitative traction.The shape of RGCs in the normal control group was round or circular,the boundary was clear,there was no obvious leaking of the light dye outside the cell,and the obvious cell protruding was seen in some of the cells.But after the traction injury of the optic nerve,the RGCs decreased continuously with time.The cell distribution is uneven,and there are a lot of clever light leakage and microglia.There was no significant difference in the RGCs density between the normal control group(right eye)and the sham operation group(only exposed to the left eye of the optic nerve).The difference was not statistically significant(P=0.228>0.05).It indicated that the exposure of the optic nerve only without traction did not cause optic nerve injury,but the number of RGCs in 1D,3D,7d and 14d decreased gradually after the optic nerve traction,and the density of the RGCs was significantly lower than that of the positive nerve.In the control group(P<0.01),the survival rate decreased(76.58%,62.47%,37.98%,16.23%,P<0.01),indicating the RGCs damage process.(2)Real-time PCR results:optic nerve traction can reduce the expression of PI3K,AKT and GSK3 beta mRNA in rat retina,while APS can promote the expression of PI3K,AKT and GSK3 beta mRNA,and the expression increases with the increase of dose.(3)Western Blot results:the effect of APS on the expression of PI3K,AKT,P-AKT,GSK3 beta and p-GSK3 beta protein:the difference between the blank group and the sham operation group was not statistically significant(P>0.05),and the expression of five indexes in the model group was significantly lower than that in the normal group and the sham operation group(P<0.01),and the APS dosage was significantly higher than that of the model group(P<0.01).Protein expression was.also increased with the increase of dose(P<0.05).The effect of APS on the phosphorylation of AKT and GSK3 beta:there was no statistical significance between the blank group,the sham operation group and the model group(P>0.05)for the analysis of the experimental results of P-AKT/AKT and P-GSK3 beta/GSK3 beta two groups(P>0.05),indicating that only the operation without traction optic nerve and traction operation did not affect the process of AKT and GSK3 beta phosphorylation in the retina tissue.APS,high dose group and model group,(P-AKT/AKT,P-GSK3 beta/GSK3 beta two group experimental results P<0.01),the difference has statistical significance,indicating that high and medium dose of APS can promote the level of phosphorylation of AKT and GSK3 beta in the retina tissue.Since PI3K/Akt transduction pathway has been clearly demonstrated to play an important role in cell survival and inhibition of apoptosis,it is concluded that Astragalus polysaccharide injection may play a role in the protection of the optic nerve by regulating the PI3K/Akt signaling pathway to inhibit the apoptosis of RGCs.Conclusion:(1)Lateral quantitative traction can establish an easy to quantify animal model of optic nerve injury,and can be used as a stable and reliable animal model for the study of the mechanism and treatment of optic nerve injury.(2)Both low medium and high dose APS have the role of up regulation of PI3K,AKT,GSK3 beta gene expression,low medium and high dose APS all have the role of up regulation of PI3K,AKT,P-AKT,GSK3 beta and p-GSK3 beta protein and expression.Meanwhile,high dose APS also has the role of promoting the phosphorylation level of AKT and beta in retina tissue.Over activation of PI3K/Akt signaling pathway can inhibit the apoptosis of RGCs,thereby playing a role in the protection of optic nerve. |
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