The Peotective Effect Of Chlorogenic Acid On Retinal Pigment Epithelial Cells Encountered Oxidative Damage

Objective:To investigate the protection of chlorogenic acid (CGA) againsthydrogen peroxide induced oxidative damage of Human Retinal Pigment EpithelialCell line(ARPE-19). Then we may provide an experimental basis for use ofchlorogenic acid for age related macular degeneration (AMD) in clinical treatment inthe future.Methods: Hydrogen peroxide(H2O2)effect on ARPE-19cell in a concentrationgradient and time gradient,then measure the cytoactive by methyl thiazolyltetrazolium (MTT) mothed to establish the best model of oxidative stress.Take themethyl thiazolyl tetrazolium(MTT) mothed to detect the proliferative effect andToxicity of the ARPE-19cells induced by chlorogenic acid(CGA).There were threegroups：normal control group，H2O2injured group，CGA treated group: CGA100μmol/L and H2O2500μmol/L.We through the The spectrophotometrically method todetect the viability of the antioxidant enzymes superoxide dismutase(SOD)，theamount of glutathione(GSH) and malondialdehyde (MDA).Measured the Apoptosisby theHoechst33258kit and flow cytometry.Results:1.ARPE-19cell was induced apoptosis by H2O2in a concentration andtime dependent，the suitable conditions for the oxidative damagemodel was500μmol/L H2O2handle for24h.2.The ARPE-19cells viability of the differentconcentration Chlorogenic acid groups compared with the blank control group wasno significant difference;Compared to the H2O2injured group，the viability ofARPE-19cell cultured with CGA and H2O2was increased．And The viability ofARPE-19injured by H2O2was greatly enhanced by100μ mol/LCGA.3.Comparedwith normal control group,the viability of the antioxidant enzymes superoxidedismutase(SOD)，the amount of glutathione(GSH) was significantly decreased, andthe amount of malondial-dehyde(MDA)was markedly increased in the H2O2injured group.While in the CGA-treated group the viability of the SOD,the amountof glutathione was increasing,and the amount of MDA was reducing,Compared with the H2O2injured group.4.The apoptosis of ARPE-19cells was obviously increasedfor the H2O2injured group.While the apoptosis was reducing in the CGA-treatedgroup.Conclusion：This tudy discovered that: H2O2can bring oxidative damage forARPE-19cell cultured in vitro,and induce the cells to apoptosis.There was noproliferative effect and Toxicity for the ARPE-19cells induced by chlorogenicacid(CGA).CGA can protect ARPE-19cell from the oxidative damage, reduce theapoptosis by H2O2。...