环孢霉素A对葡萄糖氧化酶诱导的HepG2线粒体功能损伤和细胞凋亡的保护作用

doi:10.3969/j.issn.1004-616x.2014.01.003

Carcinogenesis, Teratogenesis & Mutagenesis

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Protective effects of cyclosporine A on glucose oxidase- induced HepG2 cell apoptosis and mitochondria dysfunction

YU Wei-hua，LIU Jiang-zheng，WANG Zhao，LIU Ying，HAI Chun-xu*

(Department of Toxicology， School of Preventive Medicine， the Forth Military Medical University， Xi’an 710032， Shaanxi, China)

Received:2013-10-10 Revised:2013-11-28 Online:2014-01-30 Published:2014-01-30
Contact: HAI Chun-xu，E-mail：cx-hai@fmmu.edu.cn

Abstract

Abstract:

OBJECTIVE: To explore the protective effects of cyclosporine A on glucose oxidase-induced liver cell apoptosis and mitochondria dysfunction. METHODS：Various concentrations of GOX and CsA were applied to HepG2 cells at different time points， and measured the cell viability using MTT assay， and the appropriate doses and time points selected. Pretreatment of HepG2 cells with 10 μmol/L CsA for 2 h， then 25 U/L GOX treatment for 12 h were used as experimental group，and blank control (RPMI-1640 for 14 h)， GOX group as positive control (pretreatment with RPMI-1640 for 2 h， treatment with 25 U/L GOX for 12 h)，CsA group as negative control (pretreatment with 10 μmol/L CsA for 2 h， cultured with RPMI-1640 for 12 h). AnnexinV- FITC apoptosis detection kit was applied to assess apoptosis by flow cytometry； mitochondrial membrane permeability transition was evaluated with the GENMED MPTP fluorescent kits. Mitochondrial membrane potential was observed by means of JC-1 staining and the fluorescence intensity was measured by LSCM. The level of ATP in HepG2 cells was measured by using ATP Assay Kit； and the oxygen consumption was calculated using liquid-phase oxygen measurement system. RESULTS：MTT assay indicated that GOX decreased cell viability in dose and time dependent manners， and 10 μmol/L CsA treatment for 2 h had some protective effect. 25 U/L GOX treatment for 12 h resulted in HepG2 cells apoptosis， associated with MPTP opening， the collapse of mitochondrial membrane potential， and the reductions of the ATP level and oxygen consumption. Pretreatment with CsA could prevent structural and functional injuries of the mitochondrial， decreasing apoptosis rate. CONCLUSION：CsA could protect HepG2 cells against mitochondrial dysfunction and apoptosis.

Key words: glucose oxidase, cyclosporine A, acute liver injury, mitochondrial membrane potential, energy supply

YU Wei-hua，LIU Jiang-zheng，WANG Zhao，LIU Ying，HAI Chun-xu. Protective effects of cyclosporine A on glucose oxidase- induced HepG2 cell apoptosis and mitochondria dysfunction[J]. Carcinogenesis, Teratogenesis & Mutagenesis, doi: 10.3969/j.issn.1004-616x.2014.01.003.

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Protective effects of cyclosporine A on glucose oxidase- induced HepG2 cell apoptosis and mitochondria dysfunction

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