洛利普兰对棕榈酸作用下SH-SY5Y细胞损伤的保护作用研究

Protective Effect of Rolipram on SH-SY5Y Cell Injury Induced by Palmitic Acid

目的研究洛利普兰(Rol)对人神经母细胞瘤细胞SH-SY5Y脂毒性损伤的作用及其机制。方法实验分为正常对照(Con)组、棕榈酸(PA)组、 PA+Rol组及Rol组。采用MTT法检测细胞存活率,采用免疫荧光法测定细胞内ROS的水平,采用Western Blot检测凋亡蛋白Cleaved-caspase3的表达。结果 0.1~10μmol/L Rol对细胞存活率无明显影响,20~200μmol/L的Rol对细胞的损伤逐渐增大。MTT检测还显示10μmol/L的Rol可增加150μmol/L PA作用下SH-SY5Y细胞存活率。PA可致SH-SY5Y细胞内ROS增加,Cleaved-caspase 3蛋白表达上调,预先给予Rol可逆转上述改变。结论 Rol对PA所致细胞脂毒性损伤具有保护作用,其保护机制可能与抑制细胞内氧化应激及细胞凋亡有关。

Objective To explore the effect and mechanism of rolipram(Rol) against lipotoxic injury of human neuroblastoma cell line SH-SY5 Y. Methods The experiment was divided into control group,palmitic acid(PA) group,PA+Rol group and Rol group.MTT method was used to detect the survival rate of cells,immunofluorescence method was used to detect the level of ROS in cells,and western blot was used to detect the expression of the apoptosis protein cleaved caspase3.Results 0.1~20μmol/L Rol had no significant effect on cell survival rate,and 50~300μmol/L Rol damaged cells gradually.MTT also showed that 10μmol/L Rol could increase the survival rate of SH-SY5 Y cells incubated with 150μmol/L PA.PA could also increase ROS in SH-SY5 Y cells and upregulate the expression of cleaved caspase3 protein,which were reversed by pretreated with Rol.Conclusion Rol has a protective effect against PA-induced lipotoxicity,and its protective mechanism may be related to the inhibition of intracellular oxidative stress and apoptosis.