Effect Of Drugs On Retinal Pigment Epithelial Cell Injury Models

Age-related macular degeneration?AMD?is a retinal degenerative disease which is controlled by many factors.Current studies have shown that the pathogenesis of AMD is related to the pathology of retinal pigment epithelial cells?RPE?.In recent years,the incidence of AMD has increased year by year,and the proportion of AMD in the elderly population is also growing,which has seriously endangered the health of the elderly.Therefore,looking for the method can be effective in the treatment of AMD has attracted more attention.Due to the complex etiological factors of AMD,western treatment can not cure all the pathological characteristics of AMD,let alone solve this disease fundamentally.Therefore,to study whether there are active ingredients in traditional Chinese treatment to AMD effectively provides a new idea for the treatment of AMD.In this paper,three different induction factors of AMD models in vivo and in vitro were established through one of the most important pathogenic factors in AMD?oxidative stress of RPE cells?.Cynaroside,a traditional Chinese medicine ingredient with protective activity against oxidative damage,was proved to possess the protective effects on RPE cells injury through various models.The protective effect and mechanism of Cynaroside were further studied,which proved that Cynaroside had potential value in the treatment of AMD.This paper mainly completes the following research contents:?1?Drug-induced oxidative stress model of RPE cells was successfully established by screening different concentrations of oxidant hydrogen peroxide?H₂O₂?.The level of oxidative stress and apoptosis in RPE cells confirmed the consistency of the model with RPE cell damage in AMD disease.Cynaroside has excellent antioxidant capacity through the detection of reactive oxygen species?ROS?,superoxide dismutase?SOD?,catalase?CAT?,malondialdehyde?MDA?and glutathione?GSH?.The detection of apoptosis-related proteins in cells proves that Cynaroside also possess remarkable anti-apoptotic ability.Finally,the detection of protein related to protein kinase B?Akt?pathway in RPE cells demonstrated that Cynaroside could protect RPE cells from oxidative stress by activating Akt signaling pathway.?2?A blue-light-mediated A2E-induced RPE cell injury model was established by building a blue-light irradiation device,and the optimal light intensity and time of the blue-light model were successfully screened.The detection of Annexin V-FITC/PI double staining and H₂DCFDA staining demonstrated that Cynaroside also possess remarkable anti-apoptotic and anti-oxidative ability in this model.Futhermore,by detecting the expression of Akt,JNK and ERK pathways in RPE cells,Cynaroside was proved to regulate the physiological function of RPE cells by regulating these signaling pathways.?3?A model of retinal structural lesion induced by sodium iodate?NaIO3?in vivo in AMD mice was successfully established.The results of fundus photography in mice showed that the severe retinal lesions in this model were similar to those in AMD.HE staining and ROS detection showed that the damage of retinal tissue in this model was related to the oxidative stress of RPE cells,which objectively verified the important role of oxidative stress of RPE cells in AMD disease.Fundus photography and HE staining also showed that Cynaroside could alleviate retinal pathological changes and protect the morphological changes of retinal structure in mice by protecting RPE cells.Finally,LDH and ROS in tissues were detected to show that Cynaroside also had biological activity to RPE cells in animals.