健脾活血祛湿方通过干预AQP9表达调控线粒体细胞凋亡通路对荷H22肝癌移植瘤裸鼠的影响研究

目的观察健脾活血祛湿方通过干预水通道蛋白9(AQP9)蛋白靶点调控线粒体细胞凋亡途径对H22肝癌移植瘤裸鼠的影响。方法采用H22肝癌移植瘤裸鼠模型,随机分为空白对照组、阳性药物(CTX)对照组、健脾活血祛湿方低、中、高剂量组,分别给予相应干预后,观察裸鼠一般情况,测量移植瘤、脾脏重量、计算抑瘤率和脾指数;免疫组化检测各组肿瘤细胞AQP9表达;检测Cty-C、Caspase-3活性;TUNEL检测细胞凋亡水平。结果各组成瘤裸鼠生存状态基本良好,与空白组相比,药物高剂量组和阳性对照组抑瘤率明显增高(P<0.05),脾指数降低(P<0.05),中、高剂量组AQP9高表达且肝癌细胞呈现凋亡趋势,高剂量组的Cty-C、Caspase-3活性显著增强(P<0.05),TUNEL显示高剂量组的凋亡细胞数增多(P<0.05)。与阳性对照组相比,AQP9、Cty-C和Caspase-3的活性表达均呈剂量依赖性升高(P<0.05),以上均提示阳性对照组和药物高剂量组有效促进肝癌细胞凋亡。结论健脾活血祛湿方可诱导AQP9高表达,调节线粒体细胞凋亡通路中相关凋亡因子的表达,促进肝癌细胞凋亡,从而抑制肿瘤细胞增殖。AQP9在线粒体细胞凋亡通路中可能起着重要作用。

Effect of Jianpi Huoxue Qushi Decoction on Regulating Mitochondrial Apoptosis Pathway by Intervening AQP9 Expression on Nude Mice with H22 Liver Cancer Transplantation

Objective To observe the effect of Jianpi Huoxue Qushi Decoction on regulating mitochondrial apoptosis pathway by intervening AQP9 protein target on nude mice with H22 liver cancer transplantation.Methods The nude mice models of H22 liver cancer transplantation were randomly divided into blank control group,positive drug(CTX)control group,Jianpi Huoxue Qushi Decoction low,medium and high dose groups.After corresponding intervention,the general conditions of nude mice were observed,and the tumor graft,spleen weight,tumor inhibition rate and spleen index were measured.Immunohistochemistry method was used to detect AQP9 expression in tumor cells of each group.The activities of Cty-C and Caspase-3 were detected.TUNEL was used to detect the level of apoptosis.Results The survival of tumor nude mice in all groups was generally good.Compared with the blank group,the tumor inhibition rate in the high dose group and positive control group increased obviously decreased(P<0.05)and the spleen index decreased significantly(P<0.05).The medium and high dose groups’had high AQP9 expression and showed a trend of apoptosis of liver tumor cells.The activities of Cty-C and Caspase 3 in the high-dose group were significantly enhanced(P<0.05).TUNEL showed high dose group’s number of apoptotic cells increased(P<0.05).Compared with the positive control group,the active expressions of AQP9,Cty-C and caspase-3 were all increased in a dose-dependent manner(P<0.05),suggesting that the positive control group and the high-dose drug group could effectively promote the apoptosis of liver cancer cells.Conclusion Jianpi Huoxue Qushi Decoction can induce the high expression of AQP9,regulate the expressions of related apoptosis factors in the apoptosis pathway of mitochondrial cells,promote the apoptosis of liver cancer cells,and thereby inhibit the proliferation of tumor cells.AQP9 may play an important role in the apoptosis pathway of mitochondrial cells.