The Study Of Synthetic Diad Contained Of Lipoic Acid And Niacin On The Injury Of Retinal Pigment Epithelium By Acrolein

Purpose:To explore the protection of new synthetic diad contained of lipoic acid and niacinon the injury of human retinal pigment epithelia-19(ARPE-19) cells by acrolein andstudy the damage of rat retinal choroidal tissue by acrolein.Methods:1、 ARPE-19cells morphology was measured by HE staining. ARPE-19cellsviability and growth curve were measured by MTT assay.2、 The injuried ARPE-19cells model induced by acrolein(AC) were mediated byMTT assay and HE staining.0μmol/L,25μmol/L,50μmol/L,100μmol/L and200μmol/L AC were added into the medium of logarithmic phase ARPE-19cellsfor24hours, to detect the optimal injury concentration of AC on the ARPE-19cells and establish the injury model of the AC.3、 The apoptosis of ARPE-19cells and protective effect of diad contained of lipoicacid and niacin on the injury model of the AC were detected by flow cytometryand WesternBlot. Pretreated the logarithmic phase of ARPE-19cells by differentconcentrations of diad contained of lipoic acid and niacin such as0μmol/L、100μmol/L、150μmol/L and100μmol/L lipoic acid for24hours.Then treated itwith50μmol/L or0μmol/L acrolein for24hours and measure the ratio ofBax/Bcl-2by WesternBlot and apoptotic effect by flow cytometry.4、 The injury of acrolein on rat retina-choroid tissue was detected by HE staining.Twelve SD rats were randomly divided into control group,4W group and8Wgroup. The control group was gavaged with200μl fresh water.And the other twogroups were gavaged with200μl fresh water dissolved with2.5mg/kg/dayacrolein for4weeks and8weeks, respectively. Following the treatment,all ratswere sacrificed and the eyeballs were stained with HE technique. 5、 Statistical analysis was deteced by the statistical software SPSS13.0on variance.All the experimental data was expressed as mean±standard deviation(μ±S). If P＞0.05, the result showed no significant difference. If P＜0.05, the resultsrevealed significant difference.Results:1、ARPE-19cells were adherent to growth within24hours.They entered into thelogarithmic growth phase rapidly from the1stto6thdays,but started to decreaseafter the6thday.2、When the concentration of acrolein reached50μmol/L, we could found thenecrosis of ARPE-19cells、the accumulation of nuclear and the decreasing cellactivity of ARPE-19cells with swelling.The total number of apoptotic cells wasmuch higher than the control one,P＜0.05.3、When the concentration of diad contained of lipoic acid and niacin reached100μmol/L,it can significantly protect the ARPE-19cells which should be injuriedby50μmol/L acrolein,P＜0.05.The ratio of Bax/Bcl-2was only0.5558in the100μmol/L protecting group,but the ratio was1.3892in the injury goup,P＜0.05.4、Retinochoroidal structure was regular and RPE-Bruch membrane was continuousin the control group and4W group. The typical hill-shaped bulge and theformation of choroidal neovascularization were found in the neurosensory retinaof8W group.Conclusion:1、Acrolein can activate the mitochondrial apoptosis pathway via increasing theexpression of Bax and decreasing the expression of Bcl-2.2、New synthetic diad contained of lipoic acid and niacin blocks the mitochondrialapoptotic signal pathway to resist the injury of acrolein on ARPE-19cells throughthe inductive expression of anti-apoptotic protein Bcl-2and the inhibitiveexpression of apoptotic protein Bax.3、 Acrolein leads the injury of ARPE cells and the formation of experimental choroidal neovascularization in rat.