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[目的]观察消瘤汤含药血清对人肝癌细胞HepG2凋亡相关因子表达的影响,进一步证实该方对肿瘤细胞的抑制作用,并初步探讨其诱导肝癌细胞凋亡的作用机制。[方法]给予昆明种小鼠中药煎剂灌胃3d后,制备消瘤汤鼠含药血清,将其作用于HepG2细胞,用四氮唑盐法(MTT法)检测含药血清对HepG2细胞增殖的抑制作用,通过流式细胞技术测定肿瘤细胞的凋亡情况,应用免疫组化法检测半胱氨酸蛋白酸3(Caspase-3)、B细胞淋巴瘤2(Bcl-2)蛋白表达水平。[结果]消瘤汤鼠含药血清作用于HepG2细胞72h后,对肿瘤细胞生长有明显抑制作用。流式细胞仪检测可见含药血清组在细胞G1/G0期前出现明显的凋亡峰,即“亚二倍体峰”。凋亡率为22.3%,明显高于对照组的1.05%(P<0.05)。免疫组化结果表明Caspase-3在含药血清干预后,表达明显增加,而Bcl-2在含药血清干预后,表达明显降低。[结论]消瘤汤可能通过激活Caspase-3及抑制Bcl-2的活性,从而诱导肝癌细胞凋亡,有效地抑制肝癌细胞生长,达到抗肿瘤的作用。
[Objective] To observe the effect of Xiaoyu Tang containing serum on the expression of apoptosis-related factor of HepG2 in human hepatoma cells, and to further confirm its inhibitory effect on tumor cells and to explore its mechanism of action. [Methods] Kunming mice were drenched with traditional Chinese medicine decoction for 3 days, and the drug-containing serum of Xiaooliang Tang was prepared and treated in HepG2 cells. The proliferation of HepG2 cells was detected by MTT assay The apoptosis of tumor cells was determined by flow cytometry. The protein expression of Caspase-3 and Bcl-2 was detected by immunohistochemistry. [Result] The tumor-inhibiting serum tumor-bearing serum in HepG2 cells was significantly inhibited after 72 h. Flow cytometry showed that the drug-containing serum group showed significant apoptotic peak before G1 / G0 phase, which was “sub-diploid peak”. The apoptosis rate was 22.3%, which was significantly higher than 1.05% of the control group (P <0.05). The results of immunohistochemistry showed that the expression of Caspase-3 was significantly increased after intervention with serum, while the expression of Bcl-2 was significantly decreased after intervention with serum. [Conclusion] Xiaoli decoction may induce the apoptosis of hepatocellular carcinoma cells by activating Caspase-3 and inhibiting the activity of Bcl-2, effectively inhibiting the growth of hepatocarcinoma cells and achieving the anti-tumor effect.