中国血液净化

中国血液净化 ›› 2015, Vol. 14 ›› Issue (10): 617-621.

• 基础研究 • 上一篇    下一篇

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

王清华1,陈晶晶1,吴银锋2   

  1. 1. 湖南省娄底市中心医院血液净化中心
    2. 湖南省娄底市中心医院肾内科
  • 收稿日期:2015-06-24 修回日期:2015-07-02 出版日期:2015-10-12 发布日期:2015-10-12
  • 通讯作者: 王清华 wangqinghualoudi@126.com E-mail:wangqinghualoudi@126.com

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

  • Received:2015-06-24 Revised:2015-07-02 Online:2015-10-12 Published:2015-10-12

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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10-3 mol/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。Westernblot 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HMrSV5 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(10-3 mol/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。

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

Abstract: 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) (10-3 mol/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 NaHS (5×10- 4, 10- 3 and 5×10- 3 mol/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- 3 and 5×10- 3 mol/L) groups as compared with those in high glucose group. Conclusions H2S protects the high glucose induced injury through the inhibition of autophagy in human peritoneal mesothelial cell line HMrSV5 cells.

Key words: Human peritoneal mesothelial cells, High glucose, Cell activity, Lactate dehydrogenase, Autophagy, Autophagy related proteins