Effect Of Bushen Yangxue Mingmu Formula On Retinal Pigment Epithetial Cell Oxidative Stress And Its Mechanism

Age-related macular degeneration(AMD)is an age-related macular degenerative eye disease,which happens in people more than 50 years and serious impacts on the patient's quality of life.Pathological changes in retinal pigment epithelium-Bruch membrane-choroid blood capillary composite layer could lead to AMD.The etiology is unclear,but many studies have shown that reactive oxygen species(ROS)mediated the retinal pigment epithelium oxidative stress plays an important role in the process of damage on the pathogenesis of AMD.Bushen Yangxue Mingmu formula is a experience formula of professer Zhuang Zeng-yua,which is often used in the treatment of Retinitis pigmentosa(RP)and dry AMD that because of RPE degenerative disease.Bushen Yangxue Mingmu formula(BSYXMMF)contains Angelica sinensis,Herba epimedii(Fructus lycii,Polygala tenuifolia Willd.Studies have proved that BSYXMMF could delay the development of photoreceptor cells apoptosis in rds mice,improve the retinal mitochondrial membrane potential,reducee ARPE-19 intracellular Ca2+ concentration in vitro,resist Ca2+ increasesd after injury caused by glutamic aci.In this study we explore the effects and mechanism of BSYXMMF in protecting RPE cell damaged by oxidative stress,which could provide a new train of thought in treating RPE degeneration and dystrophy disease.Methods:To observe the effects of Bushen Yangxue Mingmu formula on oxidative stress resistance.in Retinal degeneration,and clarify the mechanism.Objective:(1)In vitro study:Animal model study:The oxidative stress model of ARPE-19 cells was induced with acrolein(75?mol/L)and different concentrations drug-contained serum of BSYXMMF were administered.ARPE-19 cells were divided into normal group(N group),model group(M group),model + normal serum group(MS group),model + low-dose BSYXMMF group(ML group),model + middle-dose BSYXMMF group(MMgroup)and model + high-dose BSYXMMF group(MH group).The activity and apoptosis rate of cell were measured by the CCK-8 method and TUNEL.(2)Animal model study:The oxidative stress model of C57 mice was induced with sodium iodate intraperitoneal injection,mice were divided in 3 groups(the model group,the lutein group and the BSYXMMF group)randomly,and then set up the normal group.To observe the changes of retinal electrophysiological activity,the structure of the retinal tissue,photoreceptor cells apoptosis and antioxidant enzymes in serum after given BSYXMMF by gavage.Result:(1)Oxidative stress intervention study in vitro showed that BSYXMMF could effect ARPE-19 cell activity~The CCK-8 proved that the effect of BSYXMMF in cell activity had relevance to the concentrations,and MMgroup and MHgroup had certain protective effect to the MS group(P<0.05),while BSYXMMF could reduce ARPE-19 cell apoptosis rate.(2)The effect of BSYXMMF on C57 mice induced by sodium iodate.?BSYXMMF could improve the electrical activity of retinal photoreceptor cells compared with litein(P<0.05).?Retinal nuclear layer cells in BSYXMMF group was more regular than that in lutein group.Outer nuclear layer in mice induced by sodium iodate was thinner than the normal.Mice in BSYXMMF group only had a layer of outer nuclear layer,and that in model group and luteinalmost disappeared.The number of photoreceptor cells in BSYMMMF group was more than in lutein group afte 14 days induced by sodium iodate(P<0.05),while the numbel of cell in inner and outer nuclear layerwere more than that in model group and lutein model.(?RPE cell nucleus in BSYXMMFG was arranged well from flat mounts of RPE-choroidal-sclera when 28 days after sodium iodate injected.? We could see a bit positive expression of 8-OhdG inRPE cell when 7 days after sodium iodate injected.While in 14 and 28 days after sodium iodate injected,8-OhdG in BSYMMMF group was weaker than that in model group lutein group.?SOD activity in BSYMMMF group is highe than in model group(P<0.05),GSH-Px activity in BSYMMMF group is highe than in model group when 14 and 28 days after sodium iodate injected(P<0.05).Conclusion:Bushen Yangxue Mingmu formula could protect RPE cell from oxidative damage,by maintaining cell morphology,decreasing the rate of cell apoptosis,improving mitochondrial DNA 8-OhdG repairing,increasing SOD and GSH-Px activity in serum.