Hydrogen sulfide protects H9c2 cardiomyocytes against high glucose-in-duced injury by inhibiting necroptosis
10.3969/j.issn.1000-4718.2016.03.001
- VernacularTitle:硫化氢通过抑制坏死性凋亡对抗高糖引起的H9 c2心肌细胞损伤
- Author:
Weijie LIANG
;
Jieyi HE
;
Wenzhu ZHANG
;
Shenglong YU
;
Jun CHEN
;
Mingcai SONG
;
Jingfu CHEN
;
Dongdan ZHENG
;
Xinxue LIAO
- Publication Type:Journal Article
- Keywords:
Necroptosis;
Hydrogen sulfide;
High glucose;
Cardiomyocytes
- From:
Chinese Journal of Pathophysiology
2016;32(3):385-391
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To study whether hydrogen sulfide (H2S) protects H9c2 cardiomyocytes against high glucose ( HG)-induced injury by inhibiting necroptosis .METHODS:The protein levels of RIP3 ( an indicator of necroptosis ) and cleaved caspase-3 were determined by Western blot .The cell viability was measured by CCK-8 assay.The intracellular le-vels of reactive oxygen species (ROS) were detected by 2’, 7’-dichlorfluorescein diacetate staining followed by photofluo-rography.Mitochondrial membrane potential (MMP) was examined by rhodamine 123 staining followed by photofluorogra-phy.The number of apoptotic cells was observed by Hoechst 33258 nuclear staining followed by photofluorography .RE-SULTS:After the H9c2 cells were treated with HG (35 mmol/L glucose) for 0~24 h, the protein expression of RIP3 in the H9c2 cells was significantly increased at 3 h, 6 h, 9 h, 12 h and 24 h, reaching the maximum level at 24 h.Pretreat-ment of the cells with 400μmol/L NaHS (a donor of H2S) or co-treatment of the cells with necrostatin-1 (Nec-1;a speci-fic inhibitor of necroptosis) considerably blocked the up-regulation of RIP3 protein induced by HG.Moreover, pretreatment with NaHS or co-treatment with Nec-1 obviously inhibited HG-induced injuries , leading to an increase in the cell viability , and decreases in the generation of ROS and MMP loss .On the other hand , pretreatment with NaHS also reduced the num-ber of apoptotic cells and the protein level of cleaved caspase-3 in the HG-treated H9c2 cardiomyocytes .CONCLUSION:H2 S protects H9c2 cardiomyocytes against HG-induced injury by inhibiting necroptosis .
