异烟肼通过ROS/Caspase-3信号通路诱导L-02细胞凋亡及槲皮素的保护作用

doi:10.3969/j.issn.1004-616x.2019.01.010

Abstract

Abstract: OBJECTIVE:To investigate involvement of the ROS/Caspase-3 signaling pathway in INH-induced apoptosis in L-02 cells,and the protective effect of quercetin. METHODS:L-02 cells were randomly divided into control and treatment groups. The latter groups were treated with INH (10 mmol/L INH),25 μmol/L quercetin (10 mmol/L INH and 25 μmol/L quercetin) and 50 μmol/L quercetin group (10 mmol/L INH and 50 μmol/L quercetin). Vitality of L-02 cells was detected by the MTT method. LDH activity in supernatant fluid was measured by colorimetric method. Apoptosis of L-02 was determined by Hoechst 33258 fluorescent staining. Mitochondria of L-02 cells,reactive oxygen species (ROS) level and mitochondrial membrane potential (△Ψm) were analyzed with fluorescent probe DCFH-DA and Rho-123. Protein expression of Caspase-3 was analyzed with western blot method. RESULTS:Compared with the controls and with L-02 cells treated with INH and quercetin,cell vitality of the INH group was significantly declined (P < 0.01). Compared with the INH group,cell vitality of the 50 μmol/L quercetin group was markedly increased (P < 0.01). Activities of LDH,cell apoptosis rate and level of mitochondrial ROS in cells of the INH group were significantly increased over the control group (P < 0.01). Activities of LDH,cell apoptosis rates and levels of mitochondrial ROS in the 25 μmol/L and 50 μmol/L quercetin groups were significantly decreased compared with that of the INH group (P < 0.05 or P < 0.01,respectively),and the effects in the 50 μmol/L quercetin group were more obvious. The mitochondrial membrane potential of the INH group was significantly lower than that of the control group (P < 0.01). The mitochondrial membrane potential of the 25 μmol/L and 50 μmol/L quercetin groups were markedly higher than that of the INH group (P < 0.05 or P < 0.01,respectively),and the 50 μmol/L quercetin group was more effective compared with the control group. Protein expression of Caspase-3 in the INH group was markedly increased (P < 0.01),compared with the INH group. Protein expressions of Caspase-3 of the 25 μmol/L and 50 μmol/L quercetin groups were significantly reduced (P < 0.05 or P < 0.01,respectively),and the 50 μmol/L quercetin group had more obvious effect. CONCLUSION:INH induced cell apoptosis in the L-02 cells and ROS/Caspase-3 signaling pathway was involved in this process.Quercetin had a protective effect against the INH-induction of cell apoptosis,the mechanism may be related to reducing ROS release,and inhibiting the ROS/Caspase-3 signaling pathway.

Key words: quercetin, isoniazid, ROS, Caspase-3, apoptosis

CLC Number:

R965

CHEN Tingyu, WANG Dongwei, ZHANG Jinbo, WANG Wei, WANG Jingtao, SHENG Yanliang, LU Chunfeng. Involvement of the ROS/Caspase-3 signaling pathway in isoniazid-induced apoptosis in L-02 cells and the protective effect of quercetin[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2019, 31(1): 53-57,68.

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Involvement of the ROS/Caspase-3 signaling pathway in isoniazid-induced apoptosis in L-02 cells and the protective effect of quercetin

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