外源性硫化氢对高糖诱导人腹膜间皮细胞氧化应激损伤的保护作用

Protective effect of exogenous hydrogen sulfide on oxidative stress injury induced by high glucose in human peritonea 1 mesothelial cells line HMrSV5 cells

目的外源性硫化氢(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5×10-5、10-4、5×10-4、10-3和5×10-3mol/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24h,MTT比色法检测细胞活力,流式细胞术检测细胞内的内活性氧(ROS)水平,检测细胞内丙二醛(MDA)含量和超氧化物歧化酶(SOD)的活性。结果与对照组比较,高糖组HMrSV5细胞活力显著降低(t=3.67,P<0.05),细胞培养液中LDH水平显著增加(t=2.94,P<0.05),ROS水平显著增加(t=4.28,P<0.01),MDA含量显著增加(t=2.84,P<0.05)而SOD活性降低(t=3.57,P<0.01)。与高糖组比较,5×10-4、10-3和5×10-3mol/L的NaHS显著抑制了高糖诱导的HMrSV5细胞活力的降低(t1=2.43;t2=3.68;t3=3.12,均P<0.05)和细胞培养液中LDH水平的增加(t1=2.83;t2=3.71;t3=3.44,均P<0.05)。与高糖组比较,10-3mol/L的NaHS显著抑制了高糖诱导的细胞内ROS(t=3.12,P<0.05)和MDA水平的增加(t=2.59,P<0.05)和SOD活性降低(t=2.61,P<0.05)。结论外源性H2S抑制了高糖诱导的人腹膜间皮细胞损伤,其机制可能与外源性H2S抑制氧化应激有关。

Objective To investigate the protective effect of exogenous hydrogen sulfide （H2S） on oxida- tivestress injury induced by high glucose in human peritoneal mesothelial cells line HMrSV5 cells. Methods HMrSV5 cells were incubated in 4.25% D-glucose （high glucose） to induce injury, and the cells were treated with the H2S donor sodium bisulfide （NariS） for 24 h. MTT assay was used to assay the cell viability. The level of reactive oxygen species （ROS） was measured by flow cytometry. Lactate dehydrogenase （LDH） in the medium, malondialdehyde （MDA） in the cells, and superoxide dismutase （SOD） activity in the cells were also measured. Results In high glucose group, cell viability and SOD activity in cells were significantly de- creased, and LDH in medium, ROS in cells, and MDA in cells were significantly increased, as compared with those in the control group. NariS reversed the effects of high glucose on HMrSV5 cells. Conclusions Exoge- nous H2S prevents oxidative stress injury induced by high glucose in human peritoneal mesothelial cells, probably through the inhibition of oxidative stress by exogenous H2S.