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目的对鱼腥草不同部位提取物诱导SGC-7901细胞凋亡及其机制进行研究。方法将鱼腥草地上茎(AS)、地下茎(SS)、叶(L)的粉末经乙醇提取得浸膏;处理SGC-7901细胞48 h后,采用Alamar blue法检测其对SGC-7901细胞增殖的影响;筛选出较高活性浸膏,光学显微镜、Hoechst33258染色观察细胞形态;DNA Ladder琼脂糖凝胶电泳检测细胞凋亡情况;Caspase-3活性检测试剂盒测定Caspase-3酶活性。Western blot检测凋亡相关因子的表达;Caspase-9抑制剂(Z-LEHD-FMK)验证浸膏调控的信号转导通路。结果各部位浸膏对SGC-7901细胞均有一定的增殖抑制活性,其中SS提取物活性最强;不同浓度的SS提取物作用SGC-7901细胞48 h后,细胞明显凋亡;DNA ladder条带也明显;Caspase-3的活性明显高于对照组;Bax、Bid、Bak、p53、Caspase-9表达上调,Bcl-2表达下调;Caspase-9抑制剂(ZLEHD-FMK)能够逆转SS对SGC-7901细胞的增殖抑制活性。结论 SS对SGC-7901细胞的增殖抑制作用最强,且是通过Caspase-9介导的线粒体凋亡信号转导通路来调控细胞凋亡的。
OBJECTIVE To study the apoptosis of SGC-7901 cells induced by extracts from different parts of Houttuynia cordata and its mechanism. Methods SGC-7901 cells were treated with Ethanol to extract the powder from the stems (AS), rhizomes (SS) and leaves (L) of Houttuynia cordata. The cells were treated with Alamar blue for 48 h The activity of Caspase-3 was detected by Caspase-3 activity assay kit. The higher active extract was screened and the morphology was observed by light microscopy and Hoechst 33258 staining. The apoptosis was detected by DNA Ladder agarose gel electrophoresis. Western blot was used to detect the expression of apoptosis related factors. Caspase-9 inhibitor (Z-LEHD-FMK) was used to verify the signal transduction pathway regulated by extract. Results The extract of each part had certain proliferation inhibitory activity on SGC-7901 cells, of which SS extract had the strongest activity. SGC-7901 cells were significantly apoptotic after treated with different concentrations of SS extract for 48 h. DNA ladder (P <0.05). Caspase-3 inhibitor (ZLEHD-FMK) could reverse the effect of SS on the expression of SGC- 7901 cell proliferation inhibitory activity. Conclusion SS has the strongest inhibitory effect on the proliferation of SGC-7901 cells and regulates apoptosis through Caspase-9-mediated mitochondrial signal transduction pathway.