加味茵陈蒿汤抑制人肝癌裸鼠皮下移植瘤生长的机制

目的:观察加味茵陈蒿汤对人肝癌裸鼠皮下移植瘤的影响。方法:建立人肝癌裸鼠皮下移植模型,随机分为生理盐水组、加味茵陈蒿汤组各8只,比较各组裸鼠瘤质量情况,流式细胞学技术检测各组瘤体细胞凋亡的分布、线粒体膜电位的分布、Caspase-3和Caspase-9的分布;透射电镜检测各组瘤体细胞形态;蛋白印迹法检测各组瘤体线粒体凋亡相关蛋白的表达。结果:加味茵陈蒿汤组的平均瘤质量均低于生理盐水组,差异有统计学意义(P<0.05)。流式细胞学技术检测显示:加味茵陈蒿汤组干预的皮下移植瘤的凋亡率高于生理盐水组、加味茵陈蒿汤组发生线粒体膜电位的比率高于生理盐水组、加味茵陈蒿汤组的Caspase-3和Caspase-9的活性均高于生理盐水组,差异均有统计学意义(P<0.05)。透射电镜显示:加味茵陈蒿汤组皮下移植瘤细胞出现细胞浆肿胀、凋亡。Western blot检测分析显示:加味茵陈蒿汤干预后的肿瘤组织中Caspase-3、Caspase-9和Bax蛋白的表达水平明显上升,Bcl-2和Bcl-x蛋白的表达水平明显降低,差异均有统计学意义(P<0.05)。结论:加味茵陈蒿汤可抑制人肝癌裸鼠皮下移植瘤生长,其作用机制可能通过活化线粒体途径诱导肿瘤细胞凋亡而实现。

Mechanism of Modified Yinchenhao Decoction Inhibiting Subcutaneous Transplantation Tumor of Human Hepatocarcinoma in Nude Mice

Objective:To observe the effect of modified Yinchenhao Decoction on subcutaneous transplantation of human hepatocellular carcinoma in nude mice.Methods:A subcutaneous transplantation model of human hepatocarcinoma was established.The rats were randomly divided into normal saline group and Jiawei Yinchenhao decoction group.The tumor mass of each group was compared.The apoptosis of each group was detected by flow cytometry.The distribution of mitochondrial membrane potential,the distribution of Caspase-3 and Caspase-9;the morphology of tumor cells in each group were detected by transmission electron microscopy;the expression of mitochondrial apoptosis-related proteins in each group was detected by Western blotting.Results:The average tumor mass of modified Yinchenhao Decoction group was lower than that of normal saline group,and the difference was statistically significant(P<0.05).Flow cytometry showed that the apoptotic rate of subcutaneous transplantation treated with modified Yinchenhao Decoction group was higher than that of normal saline group and modified Yinchenhao Decoction group.The ratio of mitochondrial membrane potential was higher than that of normal saline group and modified Yinchenhao soup.The activities of Caspase-3 and Caspase-9 in the group were higher than those in the saline group,and the differences were statistically significant(P<0.05).Transmission electron microscopy showed that cytoplasmic swelling and apoptosis occurred in the subcutaneously transplanted tumor cells of modified Yinchenhao Decoction group.Western blot analysis showed that the expression levels of Caspase-3,Caspase-9 and Bax in the tumor tissues were significantly up-regulated after the intervention of modified Yinchenhao Decoction,and the expression levels of Bcl-2 and Bcl-x proteins were significantly decreased(P<0.05).Conclusion:Modified Yinchenhao Decoction can inhibit the growth of subcutaneous transplantation in human liver cancer nude mice,and its mechanism may be induced by activating mitochondrial pathway to induce tumor cell apoptosis.