党参、甘草糖提取物对小肠上皮细胞迁移多胺信号通路的影响  

Effect of Codonopsis and Glycyrrhizae saccharide extracts on polyamine-dependent signaling pathway during cell migration in IEC-6

作　　者：李茹柳[1] 陶玉珠[1] 曾丹[1] 赵世清[2] 林传权[1] 陈蔚文[1] 

机构地区：[1]广州中医药大学脾胃研究所,广东广州510405 [2]广州中医药大学中药学院,广东广州510006

出　　处：《中国药理学通报》2016年第2期245-251,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 30772753;81173254);中央财政支持地方高校发展专项资金项目"重大难治性脾胃病防治协同创新平台"[财教(2013)338号];华南中医药协同创新中心-中医药防治脾胃病;脑病创新研究团队项目(No E1-KFD015141K03)

摘　　要：目的观察益气健脾中药党参、甘草的糖提取物对小肠上皮细胞(IEC-6)迁移过程多胺介导钾通道激活信号通路的影响,探讨党参、甘草促进胃肠黏膜损伤修复的作用机制。方法在IEC-6细胞迁移模型上,于正常或抑制多胺(加入DFMO)时,观察党参、甘草糖提取物对该信号通路的作用:(1)Western blot检测钾通道蛋白Kv1.1蛋白表达;(2)流式细胞仪检测细胞膜电位;(3)激光共聚焦显微镜检测细胞内Ca^(2+)水平;(4)Western blot检测Ca^(2+)下游指标RhoA蛋白表达。结果党参、甘草糖提取物在细胞迁移过程可以:(1)提高Kv1.1蛋白表达水平,改善DFMO所致Kv1.1蛋白表达下降;(2)提高细胞膜电位超极化水平,逆转DFMO所致细胞膜电位去极化;(3)提高细胞内Ca^(2+)水平,党参糖提取物可逆转DFMO所致Ca^(2+)水平降低;(4)提高RhoA蛋白表达水平,改善DFMO所致RhoA蛋白表达降低。结论党参、甘草糖提取物促进IEC-6细胞迁移的作用机制与其影响多胺介导钾通道激活信号通路有关。Aims To observe the effect of saccharide extracts of Yiqijianpi herb Codonopsis and Glycyrrhizae on polyamine-dependent activation of K^+ channels signal pathway during cell migration and to investigate their mechanism of promoting restoration in gastrointestinal mucosal injuries.Method The study was based on IEC-6 cell migration model.While in a normal poly amine level or poly amine was inhibited by DFMO,the effect of Codonopsis saccharide extracts and Glycyrrhizae saccharide extracts on polyamine-dependent activation of K^+ channels signal pathway during cell migration was observed.(1) K^+ channel protein Kv1.1was determined by Western blot.(2) Membrane potential was measured by Flow Cytometer.(3) Laser scanning confocal microscope was used for measuring[Ca^(2+)]cyt.(4) The expression of RhoA,which is Ca^(2+) downstream protein,was determined by Western blot.Results During cell migration,Codonopsis and Glycyrrhizae saccharide extracts could:(1) improve the expression of Kv1.1 protein and ameliorate the decrease of kv1.1 protein expression by DFMO;(2) increase membrane hyperpolarization and reverse membrane depolarization resulted by DFMO;(3) improve intracellular[Ca^(2+)]cyt,while Codonopsis could reverse the decrease of[Ca^(2+)]cyt caused by DFMO;(4) improve the expression of RhoA protein,reversing its decline caused by DFMO.Conclusion Codonopsis and Glycyrrhizae saccharide extracts can promote cell migration in IEC-6 cell,which is correlated with their effect on polyamine-dependent activation of K^+channels signal pathway.

关 键 词：党参 甘草 糖提取物 小肠上皮细胞 迁移 作用机制 

分 类 号：R-332[医药卫生] R284.1