硫化氢通过抑制自噬保护高糖诱导人腹膜间皮细胞的损伤  

Hydrogen sulfide protects the high glucose induced injury through inhibition of autophagy in human peritoneal mesothelial cells

作　　者：王清华[1] 陈晶晶[1] 吴银锋[2] 

机构地区：[1]湖南娄底市中心医院血液净化中心,娄底417118 [2]湖南娄底市中心医院肾内科,娄底417118

出　　处：《中国血液净化》2015年第10期617-621,共5页Chinese Journal of Blood Purification

摘　　要：目的探讨硫化氢(H2S)对高糖诱导人腹膜间皮细胞损伤的保护作用及其机制。方法用4.25%D-葡萄糖(高糖)和10-3mol/L的外源性H2S供体硫化氢钠(NaHS)处理人腹膜间皮细胞株HMrSV5细胞24 h,MTT比色法检测细胞活力,分光光度法测定细胞培养液中乳酸脱氢酶(LDH)水平。Western blot检测自噬相关蛋白微管相关蛋白1轻链3(LC3)、Beclin-1和p62/SQSTM1的表达。结果与对照组比较,高糖组HMr SV5细胞活力显著降低(t=4.85,P<0.05),细胞培养液中LDH水平的显著增加(t=3.49,P<0.05)。与高糖组比较,NaHS(10-3mol/L)组细胞活力显著增加(t=3.42,P<0.05)和培养液中LDH水平显著降低(t=4.02,P<0.05)。与对照组比较,高糖组HMrSV5细胞中Beclin-1(t=4.95,P<0.05)和LC3-Ⅱ(t=5.21,P<0.05)的表达以及LC3-Ⅱ/LC3-Ⅰ的比值(t=5.72,P<0.05)均显著增加,而p62的表达显著降低(t=4.67,P<0.05)。与高糖组比较,NaHS(10-3mol/L)组Beclin-1(t=4.36,P<0.05)和LC3-Ⅱ(t=4.19,P<0.05)的表达以及LC3-Ⅱ/LC3-Ⅰ的比值(t=4.83,P<0.05)显著性降低,而p62的表达显著增加(t=4.78,P<0.05)。结论 H2S保护了高糖诱导的人腹膜间皮细胞损伤,其机制可能与H2S抑制细胞自噬有关。Objective To investigate the protective effect and possible mechanism of hydrogen sulfide（H2S） on high glucose induced injury in human peritoneal mesothelial cell line HMrSV5 cells. Methods HMrSV5 cells were incubated in 4.25% D-glucose（high glucose） medium to induce the injury. The cells were then treated with a H2S donor sodium bisulfide（NaHS）（10- 3mol/L） for 24 h. Cell viability was measured by MTT assay. Lactate dehydrogenase（LDH） in cultured medium was determined by spectrophotometric method. The molecular markers of autophagy Beclin-1 and microtubule-associated protein light chain-3（LC3） and the autophagy substrates p62/SQSTM1 were determined by western blot. Results Cell viability decreased significantly and LDH level in cultured medium increased significantly in high glucose group as compared with those in control group. Cell viability became increased significantly and LDH level in cultured medium became decreased significantly in Na HS（5 × 10- 4, 10- 3and 5 × 10- 3mol/L） groups as compared with those in high glucose group. The expressions of Beclin-1 and LC3 increased significantly and the expression of p62 decreased significantly in high glucose group as compared with those in control group. The expressions of Beclin- 1 and LC3 became significantly decreased and the expression of p62 became significantly increased in NaHS（5 × 10- 4, 10- 3and 5 × 10- 3mol/L） groups as compared with those in high glucose group. Conclusions H2 S protects the high glucose induced injury through the inhibition of autophagy in human peritoneal mesothelial cell line HMrSV5 cells.

关 键 词：人腹膜间皮细胞 高糖 细胞活力 乳酸脱氢酶 自噬 自噬相关蛋白 

分 类 号：R446.6[医药卫生—诊断学]