Protective Effects Of Endogenous Hydrogen Sulfide On Beryllium Sulfate-induced 16HBE Injury And Its Mechanism

Objective: In this experiment,human bronchial epithelial cells 16 HBE were infected with beryllium sulfate(Be SO4),and endogenous hydrogen sulfide(H2S)donor sodium hydrosulfide(Na HS)and lung tissue endogenous H2 S synthase cystathionine were used to be intervened.To observe the damage effect of Be SO4 on 16 HBE cells,and to elucidate the protective effect of endogenous H2 S on Be SO4-induced 16 HBE cell injury and to explore its mechanism.Method: Human bronchial epithelial cells 16 HBE were cultured in vitro and cells were treated with different concentrations of Be SO4.Cell viability was measured by MTT assay,pretreatment of cells with H2 S donor Na HS for 6 h increased the amount of endogenous H2 S,pretreatment of cells with PPG for 6 h inhibited the expression of CSE,reduced the amount of endogenous H2 S,and subsequently treated with Be SO4.In the cells,ROS levels in the cells were measured using the fluorescent probe DCFH-DA,MDA levels were detected using the thiobarbituric acid method,and superoxide dismutase(SOD)and glutathione were detected in the cells.Peptide peroxidase(GSH-PX)levels,8-hydroxydeoxyguanosine(8-OHDG)detected by immunohistochemistry,CSE,phosphatidylinositol 3-kinase(PI3K),total Akt detected by Western blotting(t-Akt),phospho-Akt(p-Akt)protein expression levels.Immunofluorescence was used to detect the expression level of Heme oxygenase-1(HO-1)in cells,and immunofluorescence was used to detect the expression level of nuclear transcription factor(Nrf2).Results: 1.Compared with the control group,Be SO4 treatment 16 HBE cells 24 h,48h,with the increase dose of Be SO4,cell viability decreased,the difference were statistically significant(P<0.05).The half-inhibitory concentration of Be SO4 treated cells for 48 h was 270 ?M.Compared with the control group,after treatment with 16 HBE cells with different concentrations of Be SO4 for 48 hours,the intracellular ROS levels significantly increased,and the differences were statistically significant(P<0.05).When the concentration of Be SO4 was 150 ?M,the ROS levels reached maximum.2.Compared with the control group,the level of ROS,MDA and 8-OHDG were increased in Be SO4 group,Be SO4+Na HS group and Be SO4+PPG group.The H2 S content,SOD,GPx and HO-1 enzyme activity in the cells were all increased.CSE protein expression in the cell was down-regulated,the difference was statistically significant(P<0.05),Na HS intervention significantly up-regulated PI3 K,p-Akt,CSE protein,increased Nrf2 into the nucleus,increased HO-1 expression,reduced oxidative damage induced by Be SO4,PPG intervention significantly aggravated the oxidative damage of cells.Conclusion:1.Be SO4 can reduce 16 HBE cell viability and induce 16 HBE oxidative damage.2.Endogenous hydrogen sulfide protects against Be SO4-induced 16 HBE cell.3.The protective effects of endogenous hydrogen sulfide on Be SO4-induced 16 HBE cell depend on the PI3K/Akt and Nrf2/HO-1 signaling pathway activation.