中药复方“松友饮”诱导肝癌细胞凋亡的实验研究  

Experimental Research on Apoptosis of Hepatocellular Carcinoma Induced by Chinese Herbal Compound Songyou Yin

作　　者：黄修燕[1] 黄自丽[2] 许永华[2] 王志刚[1] 黄新余[1] 郑起[1] 汤钊猷[3] 

机构地区：[1]上海交通大学附属第六人民医院普外科,上海200233 [2]上海市徐汇区中心医院影像科,上海200031 [3]复旦大学肝癌研究所,上海200032

出　　处：《中华中医药学刊》2014年第2期249-251,共3页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81272401);上海市卫生局中医药科研基金项目(2010L059A);上海市卫生局中医药科研基金项目(2012QJ001A)

摘　　要：目的:研究中药复方"松友饮"对人肝癌细胞凋亡的影响及可能的分子机制,为临床应用提供理论依据。方法:采用流式细胞术、Caspase-3活性实验及Real Time PCR技术,观察2、4、8、10、20、30及40 mg/mL"松友饮"(c1、c2、c3、c4、c5、c6及c7组)对体外培养的人肝癌细胞MHCC97H凋亡的影响,并探索可能的分子机制。结果:流式细胞仪检测结果显示,同对照组相比,干预组细胞凋亡率呈时间依赖性增加,"松友饮"(c4,10 mg/mL)干预72 h后MHCC97H细胞凋亡率增加超过3倍。干预48 h后Caspase-3显著激活,Caspase-3活化呈时间依赖性。"松友饮"浓度增加,转移抑制基因1(Metastasis suppressor 1,MTSS1)抑制效果越明显,"松友饮"(c4,10mg/mL)干预48 h,MTSS1基因表达抑制率为39.0%。结论:"松友饮"诱导肝癌细胞凋亡可能与MTSS1基因表达抑制、Caspase-3活化相关。Objective： To investigate the mechanism of Chinese herbal compound Songyou Yin inducing apoptosis in human hepatocellular carcinoma （HCC）, and to provide the experimental basis of Songyou Yin in treatment for HCC.Methods ： Flow cytometry （ FCM）, Caspase - 3 experiment, and Real - Time PCR were applied to test changes of HCC treated by Songyou Yin at final concentrations of 0 （ cO group, control） ,2,4,8,10,20,30,40 mg/mL （ cl, c2, c3, c4, c5, c6, and c7 groups, respectively） for 24,48,72 h, and the underlying mechanisms were explored. SAS 8.2 software was applied for statistical analysis, and the significance was defined as P 〈 0.05. Results：MHCC97H cells were sensitive to Songyou Yin. Songyou Yin （ c4,10 mg/mL） did not cause MHCC97H cell cycle arrest at GO/G1 until 72 h treatment after the release of synchronized cultures via FCM analysis, but the apoptosis rate of treated cells was higher than that in control in a time -dependent manner throughout the period of study. The in vitro apoptosis index by FCM analysis increased up to about threefold following 72 h of treatment by Songyou Yin （c4,10 mg/mL,P 〈0.05）. We observed that Songyou Yin caused a significant apoptosis increase in MHCC97H ceils, where longer treatment times were more effective. To examine the apoptosis common effectors via the measurement of Caspase -3 in vitro with nontoxic Songyou Yin treatment （c4,10 mg/mL） ,our data revealed that Caspase- 3 was evidently activated after 48 h of treatment, accorded with the inhibited effect on metastasis suppressor 1 （MTSS1） by inhibition rate of 39.0% （P 〈 0.05 ）. Conclusion： Nontoxic herbal compound extract Songyou Yin inducing HCC apoptosis may be correlated with activation of Caspase - 3 and the inhibition effect on MTSS1 expression.

关 键 词：肝细胞癌 中药复方 “松友饮” 凋亡 

分 类 号：R285.5[医药卫生—中药学]