枸杞多糖对H2O2诱导的肾小管细胞损伤的保护作用

Protective effect of Lycium barbarum polysaccharide on renal tubular cell injury induced by HO

目的人肾小管细胞株HK-2在过氧化氢(H2O2)的作用下发生损伤,分析加入枸杞多糖(LBP)后其损伤的变化和作用机制。方法将HK-2细胞随机分成4组,每组设6个重复孔,采用不同的方法进行孵育。正常组:培养液;H2O2组:浓度为0.10 mmol/L H2O2的培养液;LBP组:浓度为100μg/mL LBP的培养液;LBP+H2O2组:提前加入LBP孵育1 h后再加入H2O2继续孵育30 min。倒置显微镜下观察每组细胞的形态变化;流式细胞仪测定每组细胞的凋亡率;测定各组细胞中丙二醛(MDA)的量和超氧化物歧化酶(SOD)的活性。结果H2O2组:与正常组比较,细胞的形态变圆,贴壁减少,细胞的凋亡率上升[(52.44±1.90)%],LBP组和正常组差异无统计学意义(P>0.05),MDA水平增加[(8.95±0.21)μmol/L]而SOD活性减少[(2.26±0.27)KU/g Pro],差异均有统计学意义(均P<0.05);LBP+H2O2组:与H2O2组比较,细胞的形态和贴壁量多均有所恢复,细胞凋亡率显著下降[(33.96±1.53)%],MDA水平减少[(4.95±0.21)μmol/L],SOD活性增加[(3.53±0.31)KU/g Pro],差异均有统计学意义(均P<0.05)。结论人肾小管细胞在H2O2的作用下发生凋亡,LBP能减少其凋亡率,这与LBP增加细胞中SOD的活性而减少MDA含量密切相关。

Objective To explore the injury of human renal tubular cell line HK-2 induced by hydrogen peroxide(H2O2),analyze the changes of injury and its mechanism after adding Lycium barbarum polysaccharide(LBP).Methods HK-2 cells were stochasticly separated into four groups,six repeat holes in each group,and incubated by different methods:normal group:culture medium;H2O2 group:culture medium+H2O2(0.10 mmol/L);LBP group:culture medium+100 ug/ml LBP;LBP+H2O2 group:incubated with H2O2 for 30 min after incubating by LBP for one hour.Morphological changes of cells in each group were observed under inverted microscope.The apoptotic rate of cells was assessed by flow cytometry.The amount of malondialdehyde(MDA)and the vitality of superoxide dismutase(SOD)in cells were determined.Results In H2O2 group,the cell morphology became round,adherence decreased,apoptotic rate increased[(52.44±1.90)%],There was no statistically significant difference between LBP group and normal group.MDA level increased[(8.95±0.21)μmol/L]while SOD activity decreased[(2.26±0.27)KU/g Pro],which had significant difference compared with the normal group(all P<0.05).In LBP+H2O2 group,the morphology and adherence of cells recovered,apoptotic rate decreased significantly[(33.96±1.53)%],MDA level decreased[(4.95±0.21)micromol/L]while SOD activity increased[(3.53±0.31)KU/g Pro],which had significant difference compared with the H2O2 group(all P<0.05).Conclusion The apoptotic rate of human renal tubular cells induced by H2O2 can be reduced by LBP,which is closely related to the increase of SOD activity and the decrease of MDA content.