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Involvement of hepatocyte nuclear factor-1b in lung bronchial epithelial cell injury induced by 2-chloroethyl ethyl sulfide
KONG Deqin, LIU Sijia, LIU Jianhao, MA Yao, MA Chengfei, ZHAO Yushun, ZHOU Jiaheng, SHI Minjie, LI Jia, LIU Jiangzheng
Carcinogenesis, Teratogenesis & Mutagenesis
2024, 36 (1):
1-8.
DOI: 10.3969/j.issn.1004-616x.2024.01.001
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.
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