Involvement of hepatocyte nuclear factor-1b in lung bronchial epithelial cell injur y induced by 2-chloroethyl ethyl sulfide
OBJECTIVE:To investigate the involvement of hepatocyte nuclear factor-1b(HNF-1b)in human lung bronchial epithelial cell injury induced by a blister agent,2-chloroethyl ethyl sulfide(CEES).METHODS:The human BEAS-2B cells were treated in culture with various concentrations(0,0.4,0.6,0.8,1.0 and 1.2 mmol/L)of CEES for 24 h.The CCK-8 method was used to detect cell viability,and cell morphology was observed under light microscope.The DCFH-DA and MitoSOX fluorescent probes were used to detect total reactive oxygen species(ROS)and mitochondrial ROS levels,respectively.The protein expression of HNF-1b was assessed by Western blot.A BEAS-2B cell line which overexpresses HNF-1b was constructed by lentiviral infection.After exposure to 1 mmol/L CEES for 24 h,cell viability was determined by the CCK-8 method;apoptosis rate was detected by Annexin V-FITC/PI double staining method;mitochondrial ROS and whole-cell ROS levels were measured by MitoSOX and DHE probes,and mitochondrial membrane potential was detected by JC-1 staining.RESULTS:After exposure of BEAS-2B to 0.6-1.2 mmol/L CEES and compared to the controls,cell viability was reduced(P<0.01),cell morphology showed damage,the levels of total ROS and mitochondrial ROS were increased(P<0.01),and HNF-1b protein expression was significantly down-regulated(P<0.01).After exposure of the cells with over-expressed HNF-1b,the cell viability was significantly increased(P<0.01),apoptosis rate was decreased(P<0.01),mitochondrial membrane potential damage was relieved,and the levels of mitochondrial ROS and total whole-cell ROS were significantly decreased(P<0.01).CONCLUSION:Exposure of the regular BEAS-2B cells to CEES reduced the expression of HNF-1b and caused extensive damage.However,overexpression of HNF-1b in the BEAS-2B cells reduced the CEES-induced cellular damage,apoptosis and mitochondrial dysfunction.These results suggest that the protective effect of HNF-1b may mediated by antioxidation,and activation of HNF-1b may be a new strategy for the treatment of blister agent toxicity.
万方数据
维普
