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目的:通过药效评价实验筛选出麦门冬汤合千金苇茎汤(简称金方)的有效部位。方法:MTT法观察不同浓度的金方提取出的8个部位对肺癌NCI-H460细胞和正常人胚肺成纤维细胞HFL-I生长增殖的影响,初步筛选有效部位;平板细胞克隆形成试验,检测有效部位对肿瘤细胞克隆形成能力的影响;同时用流式细胞仪进行细胞周期检测分析,评价有效部位对细胞周期的影响;Hoechst33258染色后在荧光显微镜下观察NCI-H460细胞凋亡的形态变化。结果:不同浓度提取各部位中,6部位对NCI-H460细胞有显著的生长抑制作用;作用72 h后,荧光显微镜下可见NCI-H460细胞核固缩,染色质凝聚等凋亡形态学特征;流式检测显示对NCI-H460细胞主要抑制在S期,并有凋亡细胞出现。结论:麦门冬汤合千金苇茎汤提取各部位中6部位对NCI-H460细胞具有细胞毒作用,可有效抑制肺癌细胞生长,并对人体正常细胞(胚肺成纤维细胞)无显著影响,可以进一步分离筛选组分及单体化合物。
OBJECTIVE: To screen out the effective sites of MaiMenDeng Decoction and QianJinWei Soup (referred to as JinFang) by the efficacy evaluation experiment. Methods: MTT method was used to observe the effects of different concentrations of Jinfang extracted eight parts on the growth and proliferation of lung cancer NCI-H460 cells and normal human embryonic lung fibroblasts, primary screening of effective sites; plate cell clone formation test, detection H4 cells were stained with Hoechst33258 and observed the morphological changes of NCI-H460 cells by fluorescence microscopy. The apoptosis of NCI-H460 cells was observed by flow cytometry (FCM). Results: Six of the extracts from different concentrations had a significant inhibitory effect on NCI-H460 cells. After 72 h, the morphological features of apoptosis of NCI-H460 cells such as nuclear condensation and chromatin condensation were observed under fluorescence microscope. The results showed that NCI-H460 cells mainly inhibited in S phase and apoptotic cells appeared. Conclusion: The combination of MaiMenDeng Decoction and QianJinWei Stem Decoction can effectively inhibit the growth of lung cancer cells and have no significant effect on the normal human embryonic lung (fibroblast) cells. Screening components and monomeric compounds can be further separated.