四君子汤含药血清上调c-Myc表达促进小肠上皮细胞迁移和增殖  

Effect of Sijunzi Tang in Promoting Migration and Proliferation of Small Intestinal Epithelial Cells by Up-Regulating c-Myc Expression

作　　者：宋厚盼[1] 李如意[1,2] 魏艳霞[1] 李鑫[1] 余黄合[2] 袁振仪[3] 蔡雄[1,3] 黄惠勇[1] 刘翠玲[4] 

机构地区：[1]湖南中医药大学中医诊断学湖南省重点实验室,长沙410208 [2]湖南中医药大学药学院,长沙410208 [3]湖南中医药大学中医方证研究转化医学湖南省高校重点实验室,长沙410208 [4]中国中医科学院中药研究所,北京100700

出　　处：《中国实验方剂学杂志》2016年第22期154-159,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：湖南省科技重大专项(2014FJ1007);湖南省教育厅优秀青年项目(15B179);湖南中医药大学博士科研启动项目(1001-017);湖南中医药大学方剂学省级重点学科开放基金项目(2015-02);湖南中医药大学中西医诊断学国家重点学科开放基金项目(2015ZYZD19)

摘　　要：目的:考察四君子汤含药血清(SJZ)对小肠上皮(IEC-6)细胞损伤后迁移、增殖及c-核蛋白类基因(c-Myc)mRNA和蛋白表达的影响,以探讨四君子汤促进胃肠黏膜损伤修复的作用机制。方法:采用大鼠制备四君子汤含药血清,Tips划痕法建立IEC-6细胞迁移模型,观察四君子汤药物血清对细胞迁移的影响;实时细胞分析仪(RTCA)检测四君子汤含药血清对IEC-6细胞增殖的影响;实时荧光定量PCR(Real-time PCR)法检测c-Myc mRNA表达;蛋白免疫印迹(Western blot)法检测c-Myc蛋白表达。结果:中体积浓度(10%)和高体积浓度(20%)的四君子汤含药血清可促进IEC-6细胞迁移(P<0.01);细胞损伤后12 h,中体积浓度(10%)和高体积浓度(20%)的四君子汤含药血清可促进IEC-6细胞增殖(P<0.01);细胞损伤后24 h和36 h,低(5%),中(10%),高(20%)体积浓度的四君子汤含药血清均可促进IEC-6细胞增殖(P<0.05,P<0.01);进一步研究发现,中体积浓度(10%)和高体积浓度(20%)的四君子汤含药血清可提高与胃肠黏膜损伤修复密切相关的c-Myc mRNA和蛋白表达(P<0.01)。结论:四君子汤修复胃肠黏膜屏障损伤,促进溃疡愈合的作用可能与其提高c-Myc mRNA和蛋白表达,促进黏膜上皮细胞的迁移和增殖有关。Objective： To study the effect of Sijunzi Tang（SJZT）-containing serum on cell migration and proliferation,c-Myc mRNA and protein expression of injured intestinal epithelial（IEC-6） cell,so as to explore the mechanism of SJZT in promoting gastrointestinal mucosal injury repair. Method： Rats were selected to prepare SJZT-containing serum,and IEC-6 cell migration model was established by tips scratch method,so as to observe the effect of SJZT on cell migration. Real-time cell analyzer（RTCA） was used to determine the effect of SJZT-containing serum on IEC-6 cell proliferation,c-Myc mRNA and protein expressions were detected by Realtime PCR and Western blot analysis,respectively. Result： The 10% and 20% SJZT-containing serum stimulated IEC-6 cell migration 8 h after cell damage（P 〈0. 01）; 10% and 20% SJZT-containing serum increased IEC-6 cell proliferation 12 h after cell damage（P 〈0. 01）. 5%,10%,and 20% SJZT-containing serum enhanced IEC-6proliferation both at 24,36 h after damage（P 〈0. 05,P 〈0. 01）. Further studies showed that,10% and 20%SJZT-containing serum increased c-Myc mRNA and protein expressions that were closely correlated with gastrointestinal mucosal injury repair（P 〈0. 01）. Conclusion： Sijunzi Tang-containing serum can repair the injury of gastrointestinal mucosal barrier,the mechanism may be related to its effect in improving mucosal c-Myc mRNA and protein expressions and promoting IEC-6 cell migration and proliferation.

关 键 词：四君子汤 胃肠黏膜损伤 细胞迁移 细胞增殖 c-核蛋白类基因 

分 类 号：R285.5[医药卫生—中药学]