罗非鱼鱼皮多肽对H_2O_2诱导的HaCaT细胞氧化应激损伤的保护作用

Protective Effect of Tilapia Skin Peptide on Hydrogen Peroxide-induced Oxidative Stress Injury in HaCaT Cells

【目的】分析罗非鱼鱼皮多肽(TSP)对H_2O_2诱导HaCaT细胞氧化损伤的保护作用及其作用机制。【方法】将HaCaT细胞随机分成空白组(细胞正常培养)、对照组(800μmol/L的H_2O_2作用24 h)和3个罗非鱼鱼皮多肽浓度组(10、20、50μmol/L TSP),应用MTT比色法检测细胞活力,DCFH-DA法检测细胞内活性氧(ROS)含量,蛋白质印迹法测超氧化物歧化酶(SOD)和谷氨酰转肽酶(GGT)含量。【结果】与空白组相比,对照组的细胞活力明显下降(P﹤0.05),细胞内ROS含量增强,GGT表达量增加,SOD表达量减少;与对照组相比,随着多肽组浓度增加,细胞活力增强,细胞内ROS含量减少,GGT表达量减少,SOD表达量增加。【结论】10~50μmol/L罗非鱼皮多肽TSP能够提高细胞活力,通过清除细胞内的ROS,抑制GGT的表达,增加SOD的表达,达到保护H_2O_2诱导的HaCaT细胞的氧化应激损伤的效果。

【Objective】To investigate the protection of Tilapia skin peptide(TSP)on H2O2-induced oxidative stress damage of HaCaT cells and its mechanism【.Method】Cells were collected and randomly assigned to the blank group(the cells were normally cultured),control group(pretreated with 800μmol/L H2O2 24 h),TSP concentration group(pretreated with 10,20,and 50μmol/L TSP for 1 h,respectively,and then treated with 800μmol/L H2O2 for 24 h).The cytotoxicity of TSP on cells was measured by MTT assay;the level of radical oxidative species(ROS)was assessed by fluorescent probe DCFH-DA;and superoxide dismutase(SOD)along with gamma-glutamyltransferase(GGT)were detected by Western blot.【Result】Compared with the blank group,there was a significant decrease in the cell viability of the control group,an increase of intra-cellular ROS content,an elevation of GGT protein expression,and a decrease of SOD protein expression.Compared with the blank group,the results showed that with the increase of TSP concentration,the relative viability of cells increased.TSP could scavenge the ROS in a dose-dependent manner,resulted in an elevation of SOD expression,and a decrease of GGT protein expression.【Conclusion】TSP(0-50μmol/L)could improve cell viability,clear intra-cellular ROS,elevate SOD protein expression and decrease GGT protein expression and thus protect HaCaT cellsfrom the oxidative stress injury induced by H2O2.