中药防风抗肝癌作用及机制的网络药理学和细胞实验研究

目的: 应用网络药理学和细胞实验,探究中药防风提取液的抗肝癌效应及可能机制,为防风在抗肝癌的临床应用提供依据。方法: 应用网络药理学方法筛选防风抗肝癌的有效成分及相应靶点,并建立"药物-成分-靶点"可视化网络;采用富集分析筛选信号通路;利用Autodock Vina软件验证效应成分与肝癌相关靶点结合力,MTT法测定细胞增殖率,流式细胞术检测细胞凋亡率,Western blot检测相关蛋白表达。结果: 通过多种数据库共筛选得到防风抗肝癌的15种有效成分和AKT1、CASP3、TP53、JUN、IL-6前5个核心靶点基因。富集分析显示细胞凋亡通路尤为集中。前5位的有效成分与核心靶点对接,结果显示其结合能均小于-20.92 kJ·mol^-1。与空白对照组比较,24和48 h时防风提取液对HepG-2细胞的抑制率随浓度升高而增大;细胞凋亡率亦升高;同时促使Caspase-3蛋白磷酸化,提高Bax/Bcl-2比值。结论: 防风提取液可抑制人肝癌细胞 HepG-2 的增殖,诱导人肝癌细胞 HepG-2凋亡,且呈剂量、时间依赖性,其作用机制可能是通过上调HepG-2细胞内Bax蛋白表达、下调Bcl-2蛋白并促进Caspase-3蛋白磷酸化发挥作用。

Network pharmacological and cellular experimental studies on the antihepatoma effect and mechanism of traditional Chinese medicine Saposhnikoviae Radix

OBJECTIVE To explore the effect and possible mechanism of Saposhnikoviae Radix extract on hepatocellular carcinoma by using network pharmacology and cell experiment, so as to provide basis for the clinical application of Saposhnikoviae Radixin to anti hepatocellular carcinoma.METHODS The effective components and corresponding targets of Saposhnikoviae Radix for anti hepatocellular carcinoma were screened by network pharmacology, and the "drug-component-target" visual network was established. The apoptosis pathway was screened by enrichment analysis. Autodock Vina software was used to verify the binding force between the effective components and hepatocellular carcinoma related targets. The cell proliferation rate was measured by MTT assay. The apoptosis rate was detected by flow cytometry, and the expression of related proteins was detected by Western blot.RESULTS Fifteen active components and five target genes of AKT1, CASP3, TP53, JUN and IL-6 of Saposhnikoviae Radix for anti hepatocellular carcinoma were screened through a variety of databases. The results showed that the binding energy of the first five active components was less than20.92 kJ·mol^-1. Compared with that of the blank control group, the inhibition rate of Saposhnikoviae Radix extract on HepG-2 cells increased with the concentration increase at 24h and 48h; the apoptosis rate also increased; at the same time, it promoted the phosphorylation of Caspase-3 protein and increased the ratio of Bax/Bcl-2.CONCLUSION Saposhnikoviae Radix extract can inhibit the proliferation of human hepatoma cell line HepG-2 and induce the apoptosis of human hepatoma cell line HepG-2 in a concentrationand time-dependent manner. Its mechanism may be related to up-regulating the expression of Bax protein in HepG-2 cells, down-regulating Bcl-2 protein and promoting the phosphorylation of Caspase-3 protein.