Role of Akt/mTOR signaling pathway in ulinastatin-induced reduction of oxidative stress injury to cardiomyocytes of rats
10.3760/cma.j.issn.0254-1416.2017.05.033
- VernacularTitle:Akt/mTOR信号通路在乌司他丁减轻大鼠心肌细胞氧化应激损伤中的作用
- Author:
Yuting WANG
;
Haiyan ZHOU
;
Hui TENG
- Keywords:
Trypsin inhibitors;
Protein-serine-threonine kinases;
Oxidative stress;
Myocytes;
Cardiac
- From:
Chinese Journal of Anesthesiology
2017;37(5):637-640
- CountryChina
- Language:Chinese
-
Abstract:
Objective To evaluate the role of protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway in ulinastatin-induced reduction of oxidative stress injury to cardiomyocytes of rats.Methods H9c2 cells were inoculated in 96-well plates at a density of 104 cells/well.The cells were divided into 5 groups using a random nomber table:control group (group C,n =12),H2O2 group (n=12),H2O2 plus ulinastatin group (group HU,n=12),H2O2 plus ulinastatin plus LY294002 group (group HUL,n=9) and H2O2 plus LY294002 group (group HL,n=9).In group H2O2,H2O2 (final concentration 200 μ mol/L) was added,and the cells were incubated for 12 h in the incubator.In HU,HL and HUL groups,ulinastatin (final concentration 50 U/ml),LY294002 (final concentration 10 μmol/L),and ulinastatin (final concentration 50 U/ml) plus LY294002 (final concentration 10 μmol/L)were added,respectively,the cells were incubated for 2 h,H2O2 (final concentration 200 μmol/L) was then added,and the cells were incubated for 12 h in the incubator.The cell survival rate was measured by methyl thiazolyl tetrazolium assay.The levels of superoxide dismutase (SOD) and malondialdehyde (MDA) were determined by using water soluble tetrazolium-1 and thiobarbituric acid colorimetric assays.Cell apoptosis was measured by flow cytometry,and apoptosis rate (AR) was calculated.The expression of Akt,phosphorylated Akt (p-Akt),mTOR,phosphorylated mTOR (p-mTOR),activated caspase-3 and activated poty-ADP-ribose polymerase (PARP) was detected by Western blot.The p-Akt/AKT and pmTOR/mTOR ratios were calculated.Results Compared with group C,the cell survival rate was significantly decreased,AR was increased,the SOD activity was decreased,the MDA content was increased,the expression of activated caspase-3 and PARP was up-regulated (P＜0.05),and no significant change was found in p-Akt/AKT ratio or p-mTOR/mTOR ratio in group H2O2 (P＞0.05).Compared with group H2O2,the cell survival rate was significantly increased,AR was decreased,the SOD activity was increased,the MDA content was decreased,the p-Akt/AKT and p-mTOR/oTOR ratios were increased,and the expression of activated caspase-3 and PARP was down-regulated in group HU (P＜0.05).Compared with group HU,the cell survival rate was significantly decreased,AR was increased,and the expression of activated caspase-3 and PARP was up-regulated in group HUL (P＜0.05).Conclusion The mechanism by which ulinastatin reduces oxidative stress injury to cardiomyocytes is related to activation of Akt/mTOR signaling pathway in rats.
