Research On Clinical Therapeutic Effect & The Mechanism Of Antagonize Apoptosis Of Zishenjianpihuayufang To DR

DR (Diabetic Retinopathy) is the most common complications of DM (Diabetes Mellitus). It belongs to "xiaokemubing" of TCM and its pathogenesis is intricate. Many complications are brought by it and it introduces blindness easily. Currently the pathogenesis is still unknown and there is little efficient and safe therapy. Presently with the deep research on cell apoptosis it has been discovered that the onset and development of DR has strong relationship with the exceptional apoptosis and pathological changes of the retinal neurocyte. Besides that the neurocyte apoptosis can not reverse. Therefore, using medicine to restrain and defer the neurocyte apoptosis in the early stage can protect the damaged retinal neurocyte with high sugar level. It is the purpose of the therapy. Chinese compound prescription is the main method of preventing and curing DR in early stage and many clinical experiences has been accumulated. Based on the TCM, the early stage DR is considered as a factor of the deficiency of Qi and Yin, blood stasis in the eyes. The principal therapy is invigorating Qi and nourishing Yin, also complementing with promoting blood circulation and restoring blood stasis. Through this method, the sight can be improved, the vision can be enhanced and the clinical development period is also delayed. However, investigations of the Chinese compound prescription are not sufficient. Most of them focus on microcirculation enhancement and blood vessel protection. The investigations about Chinese compound prescription for restraining DR neurocyte apoptosis especially in retina neurocyte are very deficient. Base on this, investigating the DR neurocyte apoptosis mechanism in early stage and searching the most efficient and convenient way of therapy is necessary.Zishenjianpihuayufang is an experiment result of Professor Yanggui Yu through many years of clinical researches. Aiming at the DR mechanism of the deficiency of Qi and Yin in the early stage, the Chinese compound prescription was developed. The main function of this prescription is invigorating Qi and nourishing Yin, also complementing with promoting blood circulation and dissolving blood stasis. The early stage experimental investigations have proved the curative effect of the prescription. It can increase eyesight, improve the ocular disease and the patients' living quality. It can also protect the retina vein and the nerval omentum of the DM rats.This research is a further investigation by using modern experimental technology based on the foundation we have got. It includes the following topics: study the cellular apoptosis inductive faaor of oxidized stressor and the inflammatory fartor's function in the DR' incidence and clinical development, discuss the pathogenesis of DR, observe the effect and the accommodate function of the Zishenjianpihuayufang to the visual acuity, the visual field observation and FFA, evaluate the efficiency and explain the validness of the micro medicine.In the side concerning the animal experiment, STZ is used to induce SD rat DM models. The method of TUNEL and immunization histochemistry was used to observe the changing of the external inductive factor of cellular apoptosis (active oxygen NOS, oxidative stress, inflammatory factor) in the peripheral blood and retina of the DM rats. The HE staining is used to observe the morphology's alterations of the DM rats' retina. The TUNEL is used to observe the neurocyte apoptosis in the rats' retina. Immunization histochemistry is used to observe the expression of the neurocyte apoptosis controlled gene Bcl-2, Bax and the apoptosis-protease Caspse-3 in the DM rats' retina in different time. Discuss the reciprocity in the neurocyte apoptosis and observe the effect of the Zishenjianpihuayufang. The above process provides the theoretic foundation of the Zishenjianpihuayufang to antagonize apoptosis and slow down DR as a multi ways. Discuss the medicine mechanism.Part I Literature reviewThrough the research of pathogenesis for DR in TCM and WM, we have learned that during the non-proliferation stage, the basic pathogenesis of DR is the deficiency of Ql and Yin and blood stasis in the eyes. Invigorating Qi, nourishing Yin, promoting blood circulation and dissolving blood stasis in the eyes are the basic therapy method. There is a close relation between neurocyte apoptosis of the retina and the occurrence and development of DR. The external inductive factor of the cellar apoptosis such as inductive oxidative stress, activating oxygen, inflammatory factor, the cell-control gene Bel-2, Bax and the apoptosis-protease Caspase-3 play an important role in the neurocyte apoptosis. Chinese medicine has the function of antagonize apoptosis and Zishenjianpihuayufang \s one of them.Part II Research of the curative effect and the mechanism of Zishenjianpihuayufang to DR during the non-proliferative DR.1. PurposeResearch the effect of the neurocyte apoptosis induced factor such as oxidative stress, active oxygen and inflammatory factor and the influence of Zishenjianpihuayufang on them. Observe the therapeutic effect of Zishenjianpihuayufang to visual function, ocular fundus pathological changes, FFA and the visual field for the NPDR patients who are deficient in both Qi and Yin. Investigate Zishenjianpihuayufang's clinic therapeutic effect and mechanism.2. Method53 patients suffering from Deficiency of Qi and Yin type NPDR are collected. 23 of them belong to the therapeutic group, taking Zishenjianpihuayufang orally. 30 of them in the compared group are treated only by reducing the blood sugar level. 20 patients are in the normal compared group. 22 cases are PRD patients. For all the patients the TNF-a, IL-6, SOD, MDA, GSH-Px and NOS of the peripheral blood are detected. Besides that the vision field and FFA inspect are also been done for the NPDR patients.3. Result3.1 Normal group, NPDR, PDR group SOD, MDA, GSH-Px, NOS, TNF-a, IL-6 levels3.1.1 SOD, MDA, GSH-Px? Compared with the normal group, for the NPDR and the PDR group the SOD, GSH-Px reduced and the difference is notable (p<0.01). MDA increased and the difference is notable (p<0.01)? Compared with the NPDR group, for the PDR group, SOD and GSH-Px reduce and the difference is notable (p>0.05). MDA increase and the difference is notable (p<0.01)3.1.2 NOS result? Compared with the normal group, for the NPDR group, the NOS rise and the difference is notable (p<0.05).? Compared with the normal group, for the PDR group, the NOS rise and the difference is notable (p<0.05)? Compared with the NPDR group, for the PDR group, the NOS rise and the difference is notable (p<0.05)3.1.3 TNF-?, IL-6 result? Compared with the normal group, for the NPDR and PDR group, the IL-6 and the TNF-a rise, the difference is very notable (p<0.01)? Compared with the NPDR group, for the PDR group, IL-6 and TNF-a increase, the difference is very notable (p<0.05)3.2 Influences of Zishenjianpihuayufang on Deficiency of Qi and Yin type NPDR patients' eyesight and their ocular fundus.? Sight raise : Comparison between the therapy group and the control group has notable difference (p<0.01)? Ocular fundus improvement: Comparison between the therapy group and the control group has notable contrast (p<0.01)3.3 Influences of Zishenjianpihuayufang on FFA of Deficiency of Qi and Yin type NPDR patients.? Om: The difference in A-RCT, microangioma' s number, macula lutea microangioma 's transudation, nonperfusion's area is not notable (p>0.05).? 3m> Therapy group: A-RCT shorten, macula lutea microangioma 's transudation is alleviated. It has notable contrast (p<0.05).Non-perfusion's area and microangioma' s number has no notable changes (p>0.05).> Control group: A-RCT, macula lutea microangioma's transudation, non-perfusion's area, microangioma number increased. However, without notable contrast (p>0.05)3.4 Influence of the Zishenjianpihuayufang on Deficiency of Qi and Yin type NPDR's visual field? Om: The comparison of MS, MD, LV, CLV, SF between therapeutics group and control group has no statistic sense (p>0.05).? 3m> Therapy groups: MS (p=0.000), MD (p=0.049), LV (p=0.039). There is notable contrast before and after the treatment. The comparison of CLV and SF has no notable contrast (p>0.05).> Compared group: During 3 months of the observation, MS reduced. The contrast is notable (p=0.008). MD, LV, CLV and SF have no notable difference.> Comparison between the therapy group and the control group: MS and MD have notable contrast.MS (p<0.05), MD (p<0.01).3.5 Influences of Zishenjianpihuayufang on Deficiency of Qi and Yin type NPDR patients' SOD, MDA, GSH-Px, NOS, TNF-a and IL-6 of the peripheral blood.? Om: Therapy group and control group's SOD, GSH-Px, MDA.NOS, TNF-a, IL-6 comparison before and after treatment don't exist notable contrast (p>0.05)? 3m> Therapy group: SOD (p=0.048) and GSH-PX (p= 0.044) increased, NOS, MDA reduced (p=0.000). TNF- a * IL-6 level reduced and the difference is notable.> Control group: During 3 months of the observation ,SOD, GSH-PX, MDA, NOS, TNF- a and IL-6 have no notable difference (p>0.05) .> Comparison between the therapy group and the control group: SOD (p= 0.007) - GSH-PX (p= 0.000)? MDA (p= 0.002). The notable difference exists.4 Conclusion4.1 DR patients have an imbalance system of oxidization and anti-oxidization. Oxidative stress is aggravated. Activity of NOS which possess neurocyte toxicity increased. Level of intense phlogosis cellular factor TNF-a, IL-6 rise. Above factors including DR's serious degree and the different stage had a close relationship in between.4.2 Zishenjianpihuayufang can improve the sight and the ocular fundus of the patient in the early stage. It can also increase MS, MD, LV, reduce A-RCT and improve the macula vessel's transudation status. It has good effect in improving the ocular fundus and increasing the sight.4.3 Using Zishenjianpihuayufang, imbalance of oxidization and inoxidation systems have been accommodated. It keeps NOS active, reduces its nerval toxicity on DR, reduces TNF-a and IL-6 level of the inflammatory factors and alleviates retinal damage. Thereby vision can be improved.Part III Retinal nerve cellular apoptosis mechanism of the STZ Inductive DM rats & research of Zishenjianpihuayufang's antagonizing apoptosis function.1. PurposeObserve the histomorphology's alterations and its cellular apoptosis mechanism of the retina through STZ inductive SD rats DM model. Explore the multitarge effect and the nerve protection mechanism of the Zishenjianpihuayufang to the cellular apoptosis signal in the conductive channel through the comparison of Zishenjianpihuayufang and Doxium as comparative therapy. Discuss Chinese compound prescription as a multi ways to antagonize apoptosis and protect retina neurocyte.VI2. Method2.1 Observision peripheral blood oxidative stress, inflammatory factor and retinal histomorphology.95 SD rats of SPF were used. 1 5 of them are taken out at random as normal group. Another 80 rats are divided into 4 groups in average: Model group, low dose of Chinese medicine group, high dose of Chinese medicine group and Doxium group. The models were made by abdominal injection of STZ 60mg/kg with empty stomach. 10 days after that, the course is being recorded. The animals are anesthetized to died group by group each 1, 3, 6 months afterward with daily intragastric administration. SOD, GSH-Px, MDA, NOS, TNF-a, IL-6 were surveyed . Eyeballs were taken as histomorphology of retina with HE staining.2.2 observision retinal neurocyte apoptosis by TUNELAnimals were anesthetized to died each 1, 3, 6 months in order to take out their eyeballs. The retina pathological section was done. The TUNEL (terminal deoxynucleotidyl transferase mediated dUTP-digoxigenin nick end labeling) was used to observe each DM rats' retinal cellular apoptosis condition, and to observe the antagonistic effect of Zishenjianpihuayufang2.3 ImmunohistochemistryEach 1, 3, 6 months, anesthetize the rats and take their eyeballs away for retinal pathological section. Observe DM rats' retinal expression of the Bcl-2, Bax, Caspase-3, NOS, TNF-a in different time by using immunohistochemistry method.3. Result3.1 The condition of SD rats' phramoid percentage and their death:The average blood sugar of the SD rats is higher than 16.65mmol/L That means the modeling is successful. The first phramoid percentage is 81.05% and the second one is 96.84%. There are no died rats during the modeling process. Within 6 months of observation, there are no dead rats in the model group. But there is one died in each of the high and low dose of Chinese medicine group separately causing by intragastric administration.3.2 Observation of general state of health of every rats group? Normal group: Spirit is normal. The fur is velvet. The movement is agile. The resistance is normal.? Modeling group: They displayed ennui, sag vigor, wilt hair, emaciation, extrado, less hair with knot. The chest and back can be seen their skin. Every rats in all DM group showed wet, watery sedes . The "3more and 1 less" phenomena (eat more, drink more, urinate more, and losing weight) is very evident. The blood sugar is persistent high.? Chinese medicine group: The "3more and 1 less" phenomena is improved distinctly. The level of blood sugar is reduced. Those symptoms displayed above are lighter than Model group and the Doxium group. The difference is remarkable (p<0.05)? Doxium group: in the symptom improvement aspect, the Doxium group is better than the model group but worse than the Chinese medicine group. In the 3rd month and the 6th month, the cataract appearance in the high dose Chinese medicine group is much less than the model group and the Doxium group. The difference is very remarkable.3.3 Change of Retinal histomorphology:? Normal rats showed clear configuration in each retinal layer in different times. The layers are clear and the arrangement is orderly. For the neurocyte in different levels, the size is uniformity, the density is equal and the arrangement is uniform.? For the model group, at the end of the 1st month, there is no visible histomorphology alteration in the retina. At the end of the 3rd month, the arrangement of the neurocyte was disorderly and sparse. The number of the neurocyte reduced, Vacuole-like transformed, karyopycnosis, a parcel of hypochromatosis, exterior and interior nuclear layer became thinner, the collocation was disorderly and the staining was aniso- . At the end of the 6th month, the above phenomena worsen. The rats had edema in their retinal nerve fiber layer and it turned thicker. The arrangement is very sparse andderangement in the outer segment.? In the above three times, the retinal histomorphology alteration in the Ooxium group is much less than the model group but more than the Chinese Medicine group. The retinal histomorphology alteration of the high dose Chinese Medicine group is less than that of the low dose Chinese Medicine group.3.4 Retinal neurocyte apoptosis? At the end of the 1st month: no positive apoptotic karyon was detected in the normal control group. In the model group the retinal ganglion cell layer and its internal nuclear layer showed a few positive apoptotic karyon . In low dose Chinese medicine group and Doxium group retinal ganglion cell layer and their internal nuclear layer showed scattered positive apoptotic karyon .? At the end of the 3rd month: scattered positive apoptotic karyon was detected in the ganglion cell layer in normal group. In the model group the ganglion cell layer and its internal nuclear layer showed a few positive Karyon . In high dose Chinese medicine group the ganglion cell layer and its interior layer were detected scattered positive Karyon . The ganglion cell layer and its internal nuclear layer showed a few positive Karyon in low dose Chinese medicine group . In Doxium group the ganglion cell layer showed many positive Karyon and internal nuclear layer had a few positive Karyon.? At the end of the 6th month: The normal control group's internal nuclear layer showed scattered positive cells. For the other groups their positive cell number increased more.3.5 Peripheral blood SOD, GSH-Px, MDA, NOS,TNF-a, IL-6's alteration:3.5.1 Comparison of peripheral blood SOD, CSH-PX and MDAAt the end of the 1st month: compared with the normal group, all other groups have the following changes: SOD decreased, MDA ascended. The difference is notable (p<0.01). Compared with the model group, all other groups have the following changes: SOD and GSH-Px increased, MDA reduced. The difference is remarkable. Comparison between different therapy groups showed that the difference of SOD and MDA is notnotable. Doxium group, low dose Chinese medicine group and high dose Chinese medicine group showed notable contrast (p<0.01) in GSH-Px.At the end of the 3rd month: Compared with the normal group, all other groups have the following changes: There is no difference in SOD level for high dose group. For other groups the SOD level decreased. The difference is remarkable. (p<0.01). CSH-Px level of all other groups are lower than the normal group. The difference is remarkable. (For Chinese medicine high dose group: p<0.05. For other groups p<0.01). The MDA level increased for the model group and the Doxium group. The difference is remarkable (p<0.01). MDA level keep the same for Chinese Medicine groups. Compared with the model group, all other groups have the following changes: For all the therapy groups, the SOD and GSH-Px level are higher than the model group. The difference is remarkable. The MDA level decreased for the Chinese Medicine group. The difference is remarkable (p<0.01). For Doxium group the comparison result doesn't make sense. Comparison between the therapy groups: there is no remarkable difference in the SOD level between the Chinese Medicine groups and the Doxium group. But the GSH-Px level of Doxium group is lower than the Chinese Medicine high dose group (p<0.01). The SOD level of high dose Chinese Medicine group is higher than the low dose group. The difference is remarkable. There is no remarkable difference in the MDA level.At the end of the 6th month: Compared with the normal group, the SOD level has no difference in the high dose level and is lower for the other groups. The GSH-Px level is lower than the normal group for all the other groups. The comparison result of the MDA level between Chinese Medicine groups and the normal group dose not make sense. Compared with the model group, the SOD and GSH-Px level increased for all the therapy groups but the MDA level decreased. The difference is remarkable. Compared between the therapy groups, the high dose Chinese Medicine group has a higher SOD and GSH-Px level than others but the MDA level is lower. The difference is remarkable (p<0.01). There is no notable difference between the low dose Chinese Medicine group and the Doxium group.3.5.2 Comparison result of IL-6 levelAt the end of the 1st month, the difference between the Chinese Medicine groupand the normal group is not remarkable, the difference between the model group and the normal group is remarkable (p<0.01), the difference between the Doxium group and the normal group is remarkable (p<0.01). Compared with the model group, the IL-6 level of all other groups reduced (p<0.01). Compared with the Doxium group, the IL-6 level of the high dose Chinese Medicine group reduced (p<0.01). There is no difference between the Chinese Medicine groups.At the end of the 3rd and the 6th month: The IL-6 levels of all therapy groups are higher than the normal group (p<0.01) but lower than the model group (p<0.01). The level of the Chinese Medicine group is excelled than the Doxium group at the end of the 3rd month. The comparison result among all the other therapy groups makes no sense.3.5.3 Comparison result of TNF-? levelAt the end of the 1st month: There is no remarkable difference between the high dose group and the normal group. For other groups the TNF-a level is higher than the normal group.At the end of the 3rd and the 6th month: The TNF-a level of all other groups is higher than the normal group. The difference is remarkable (p<0.01). There is a remarkable difference between the Chinese Medicine group and the model group. The comparison result of the Doxium group and the model group has a remarkable difference at the end of the 6th month. The TNF-a level of high dose Chinese Medicine group is lower than all the other groups at any time.3.5.4 Comparison result of the NOS level:For all the other groups the NOS level is high than the normal group at any time. The difference is remarkable. The NOS level of Chinese Medicine group is lower than that of the model group at any time. At the end of the 1st month, the level difference of the Doxium group and the model group has no statistical significance. Compared between the therapy groups, the NOS level of Chinese Medicine group is lower than the Doxium group. At the end of the 3rd month, the curative effect of the low dose Chinese Medicine group decreased and is almost the same with the Doxium group. But for the high dose Chinese Medicine group the curative effect is much better and the differenceis remarkable. The NOS level of the high dose group is lower than that of the low dose group.3.6 Expression of retinal NOS, TNF-aThe model group: At the end of the 1st month, the positive caryon were detected in the retina in model group . At the end of the 6th month, both the pigmentation's intensity and the number of positive caryon increased. The main expression site was located in ganglion cell layer, internal layer and external plexiform layer's cytomembrane and cytolymph.High dose Chinese medicine group: At the end of the 1st month, unremarkable positive cell expression was displayed in each layer. At the end of the 3rd month, retinal ganglion cell layer and inner nuclear layer showed scattered positive cell expression. External plexiform layer showed positive cell expression. At the end of the 6th month, positive cell was seen in each retinal layer.Low dose Chinese medicine group and Doxium group: At the end of the 1st month, some positive cell expression could be detected in each layer. At the end of the 3rd month, The number of the positive cells and Pigmentation increased. At the end of the 6th month, numerous positive cells could be seen in each retinal layer.For the normal group, internal nuclear layer scattered positive cell expression could be seen merely at the end of the 6th month.3.7 Expression of retinal Bel -2,Bax, Caspase-3:3.7.1 Bel-2:Normal group: The expression's location and intensity of the positive cells are identical in any times. The main location was retinal vessel wall, scattered retinal ganglion cell layer, internal nuclear layer, etc.Model group: At the end of the 1st month, positive cells were mainly located in vessel wall. It was intenser than the normal group's pigmentation, but inferior to the therapy groups'. At the end of the 3rd and the 6th month, positive cell had enlarged into every vessel wall, ganglion cell layer, external plexiform layer. The stain enhanced.High dose Chinese medicine group: At the end of the 1?? month, the Bel -2 positivecells were mainly located in vessel wall and ganglion cell layer. The number was higher than the other DM groups. At the end of the 3rd month, the main location was the retinal vessel wall, the ganglion cell layer and the external plexiform layer. Some of them were located in internal nuclear layer and the stain enhance. At the end of 6th month, positive cell expression number and the stain intensity increase more and enlarge into the internal nuclear layer.Low dose Chinese medicine group and Doxium group: At the end of the 1st month, positive cells mainly located in external plaxiform vascular wall. The number of positive cells was higher than that of the model group. At the end of the 3rd month, the positive cells distended to gangliocyte layer. Scattered positive cells could be found in the external plsxiform. At the end of the 6th month, The number of the positive cells increased and the pigmentation intensed.3.7.2 BaxNormal group: At the end of the 1st month there is scattered positive cell in the internal nuclear layer. At the end of the 3rd month, there are scattered positive cell in the ganglion cell layer and a few in the vessel walls. At the end of the 6th month, stain increased and the location of expression is the same with the 3rd month.Model group: At the end of the 1st month and the 3rd month, there are numerous positive cells located in ganglion cell layer and external plexiform layer. Some of them were located in the internal nuclear layer and vessel wall. At the end of the 6th month, positive cells increased further. Stain enhanced and the expression located in every layers of the retina.High dose Chinese Medicine group: At the end of the 1st month, the main positive expression is in the vessel wall and scattered ganglion cell layer. The stain intensity was inferior to the model group and other groups. At the end of the 3rd month, expression had enlarged into internal nuclear layer. At the end of the 6th month, the expression ambit was unchanged. The positive cell number increased. The stain enhanced but still inferior to model group and other taking medicine group.Low dose Chinese Medicine group and the Doxium group: At the end of the 1st month, the main expression of the positive cells is in the vessel wall, the scatteredganglion cell layer and the internal nuclear layer. At the end of the 3rd and the 6th month, the positive cells enlarged into the external plexiform layer. The number of the positive cells increased and the stain enhanced.3.7.3 Csapase-3:Normal group: There was no positive expression at any time.Model group: At the end of the 1st month, the positive cell was situated in ganglion cell layer and scattered in the internal nuclear layer. At the end of the 3rd month, it was mainly located in ganglion cell layer. INL had a few positive cells. At the end of the 6th month, there was a large amount of positive cell in the CCL and the INL.High dose Chinese medicine group: At the end of the 1st month, no remarkable expression could be seen. At the end of the 3rd month, the CCL and the INL were beginning to show positive cell expression. At the end of the 6th month, the expression ambit was almost the same as that of the 3rd month. The number of positive cell increased, the stain increased but inferior to the model group and other therapy groups.Low dose Chinese medicine group: At the end of the 1st month, positive cell was mainly located in the CCL. At the end of the 3rd month, the expression was mainly located in the CCL.there is some positive cell in the INL. At the end of the 6th month, much positive cell expression could be detected in the CCL and the INL.Doxium group: At the end of the 1st month, it was mainly situated in the GCL and scattered in the INL. At the end of the 3rd month, positive cell expression ambit was unchanged but the number increased and the stain intensed. At the end of the 6th month, there is much positive cell expression in the GCL and the INL. The intension and the number of the stain is higher than the Chinese medicine groups. 4 Conclusion 4.1STZ inductive adult SD rats showed high rate of DM phragmoid. After 6 months of modeling, the DM rats' condition of consciousness, color, physical appearance, gesture and the "3more and 1 little" symptom were corresponding to human DR pathogenesis, which mainly showed in lung, spleen, and kidney. The histopathology changing of theretina reflected the pathogenesis changing of the NPDR. This provides a good animalmodel for Zishenjianpihuayufang to prevent and cure DR.4.2From the 2nd month, the retina nerve cell of the STZ induced DM rats began to apoptosis. With the prolonging of the diseases course, the number of the apoptosis cells increased.The oxidation - antioxidation imbalance and the oxidative stress intensed, NOS activity increased. The inflammatory factor TNF- a and IL-6 level increased.The cell controlled gene and the protease changed. The Bcl-2/Bax expression unbalanced. With the prolong of the diseases course the Caspase-3 expression of the retina increased. 4.3The Zishenjianpihuayufang can improve the "3 more and 1 little" symptom and reduce the blood sugar and the probability of the cataract. The relationship between the quantity and the effect could be detected.The Zishenjianpihuayufang can degrade peripheral blood MDA, SOD and CSH-Px level, raise SOD, GSH-Px level and reduce the expression of IL-6 and NOS in the retina. Base on that it can relieve the oxidation stress reaction and reduce the irritation from the inflammatory factor to the organism and the retina. This could be one of the mechanisms of the Zishenjianpihuayufang to restrain and slow the nerve cell apoptosis through reducing the induced factor of the nerve cells.The Zishenjianpihuayufang can increase the Bcl-2 and Caspase-3 expression quantity of the SD rats' retina and reduce the expression quantity of the Bax. This may be one of the mechanisms that it can restrain the nerve cell apoptosis of the retina.The Zishenjianpihuayufang can improve the histopathology change of the retina. It can also defer and restrain the nerve cell apoptosis. From this the relationship between the quantity and the effect could be found.