JNK信号通路在异烟肼诱导L-02细胞凋亡中的作用及槲皮素干预作用

The role of JNK signaling pathway in INH induced L-02 apoptosis and intervention of quercetin

目的探讨JNK信号通路在INH诱导L-02细胞凋亡中的作用及槲皮素的干预作用。方法建立INH肝细胞L-02损伤模型,实验分为未处理对照组、INH模型组、槲皮素低剂量及高剂量组。应用MTT法检测L-02细胞存活率,利用Hoechst 33258荧光染色法评价细胞凋亡情况,采用Western blot法检测细胞中Bcl-2、Bax、Caspase 3、Caspase 9和JNK蛋白的表达。结果与未处理对照组相比,INH模型组细胞存活率明显降低,细胞凋亡率明显增高,细胞中Bcl-2蛋白表达明显降低,而Bax、Caspase 3、Caspase 9和p-JNK蛋白表达明显增加。槲皮素能明显增加细胞存活率,明显降低了细胞的凋亡率,槲皮素高剂量组细胞Bcl-2蛋白表达明显增高,而Bax、Caspase 3、Caspase 9和p-JNK蛋白表达明显减少。结论 JNK信号通路在INH诱导L-02细胞凋亡中发挥了重要作用;槲皮素对INH诱导L-02细胞凋亡有保护作用,可能与其调节凋亡相关蛋白的表达,抑制JNK信号通路有关。

Objective To investigate the role of JNK signaling pathway in INH induced L-02 apoptosis and intervention of quercetin. Methods The injury model of liver L-02 cells induced by INH was established. The cells were divided into untreated control group,INH model group,low dose and high dose quercetin groups. Cell survival rate was detected by MTT method. Apoptosis was detected using Hoechst 33258 fluorescent staining; The protein expressions of Bcl-2,Bax,Caspase 3,Caspase 9 and JNK in cells were determined by western blot method. Results Compared with untreated control group,the cell survival rate was significantly lower,the apoptotic rate of INH model group was obviously higher,the Bcl-2 protein expression was significantly lower while Bax,Caspase 3,Caspase 9 and p-JNK protein expressions were markedly increased in INH model group. The cell survival rates of quercetin groups were significantly increased,the apoptotic rates were significantly decreased,the Bcl-2 protein expression was obviously higher,and Bax,Caspase 3,Caspase 9 and p-JNK protein expressions were decreased significantly in high dose quercetin group. Conclusion JNK signaling pathway played an important role in INH induced L-02 apoptosis; Quercetin could obviously protect L-02 cells from apoptosis induced by INH,and the mechanism may be related to regulate the expressions of apoptosis related proteins and to inhibit JNK signaling pathway.