半夏泻心汤含药血清对胃癌微环境中BMSCs生长增殖的影响  

Effect of Serum Containing Banxia Xiexin Tang on Growth and Proliferation of Bone Mesenchymal Stem Cells in Gastric Carcinoma Microenvironment

作　　者：杨柏林[1] 刘喜平[1] 崔国宁 董俊刚[1] 明海霞[1] 辛宝[2] 吴高峰[1] 

机构地区：[1]甘肃中医药大学基础医学院,兰州730000 [2]陕西中医药大学,陕西咸阳712046

出　　处：《中国实验方剂学杂志》2016年第22期97-102,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81260525);甘肃省属高校基本科研业务费专项资金项目(BH-2011-068)

摘　　要：目的:观察半夏泻心汤及不同配伍药组含药血清对胃癌微环境中骨髓间充质干细胞(BMSCs)生长增殖的影响,并探讨其作用机制。方法:设BMSCs常规培养为空白组;模型组采用transwell小室将人胃癌BCG-823细胞与大鼠BMSCs非接触共培养建立胃癌微环境;半夏泻心汤及不同配伍含药血清组(全方组、辛开组、苦降组、甘补组)在上室内加入体积浓度为10%的相应配伍药含药血清。倒置显微镜观察BMSCs细胞形态、噻唑蓝(MTT)比色法观察BMSCs的增殖率,流式细胞术(FCM)检测BMSCs的细胞周期,蛋白免疫印迹法(Western blot)检测原癌基因(c-Myc)和端粒酶逆转录酶(TERT)蛋白的表达,实时荧光定量PCR检测c-Myc和TERT mRNA的表达。结果:全方组、辛开组、苦降组、甘补组含药血清在4,5,6,7 d对BMSCs的增殖有明显抑制作用,全方组明显优于甘补组(P<0.05);第7天的检测结果显示全方组、辛开组、苦降组、甘补组含药血清均可降低c-Myc和TERT表达水平,升高G1期细胞比例,降低S期细胞比例,全方组明显优于甘补组(P<0.05)。结论:半夏泻心汤及不同配伍药组含药血可抑制胃癌微环境中BMSCs的异常增殖,半夏泻心汤全方为最佳配伍。Objective： To observe the effect of serum containing Banxia Xiexin Tang and its different combinations with other herbs on growth and proliferation of Bone Mesenchymal Stem Cells（BMSCs） in gastric carcinoma microenvironment. Method： To BMSCs were routinely cultured as blank group. In this study,a coculture system of BMSCs and gastric carcinoma cell line BCG-823 was established by using transwell chamber as model group. Groups of serum containing of Banxia Xiexin Tang and its different combinations with other herbs（Quanfang group,Xinkai group,Kujiang group and Ganbu group） were added with the corresponding drugcontaining serums at the concentration of 10%. The phase-contrast microscope was used to the morphology of BMSCs cell,methylthiazolyldiphenyl-tetrazolium bromide（MTT） method was adopted to observe the proliferation rate of BMSCs,flow cytometry（FCM） was applied to test BMSCs cell cycle,c-Myc and expression of telomerase reverse transcriptase（TERT） protein were detected by Western blot,fluorescence Real-time PCR test was used to detect c-Myc and TERT mRNA expressions. Result： Quanfang group,Xinkai group,Kujiang group and Ganbu group medicated serum at 4,5,6,7 d had a significant inhibitory effect on the proliferation of BMSCs,Quanfang group was superior to Ganbu group（P〈 0. 05）,the detection results at the 7thday show that Quanfang group,Xinkai group,Kujiang group and Ganbu group medicated serum can decrease c-Myc and TERT expressions,elevate G1 phase cell proportion,and reduce the proportion of S phase cells,and Quanfang group was superior to Ganbu group（P 〈0. 05）. Conclusion： The serum containing Banxia Xiexin Tang and its different combinations with other herbs inhibit the abnormal proliferation of BMSCs in gastric carcinoma microenvironment,the overall prescription of Banxia Xiexin Tang shows the best compatibility.

关 键 词：骨髓间充质干细胞 半夏泻心汤 胃癌微环境 细胞增殖 原癌基因 端粒酶逆转录酶 

分 类 号：R285.5[医药卫生—中药学]