四君子汤含药血清对IEC-6细胞增殖及RhoA、p21、Cdk2表达的影响  

Effects of Sijunzi Decoction-medicated serum on IEC-6 cell proliferation and RhoA, p21 and Cdk2 expressions

作　　者：涂小华 杨光勇 杨欣 邓颖 徐萌萌 魏砚君 万亿 王慧 TU Xiao-hua;YANG Guang-yong;YANG Xin;DENG Ying;XU Meng-meng;WEI Yan-jun;WAN Yi;WANG Hui(School of Basic Medical Sciences,Guizhou University of Traditional Chinese Medicine,Guizhou 550025,China)

机构地区：[1]贵州中医药大学基础医学院,贵州贵阳550025

出　　处：《中成药》2022年第7期2125-2131,共7页Chinese Traditional Patent Medicine

基　　金：国家自然科学基金项目(81760864);贵州省教育厅青年科技人才成长项目(黔教合KY字[2017]173);贵州中医药大学博士启动基金(20160035)。

摘　　要：目的 探究四君子汤含药血清对IEC-6细胞增殖及Rho GTP酶(RhoA)、p21、细胞周期蛋白依赖性激酶2(Cdk2)表达的影响。方法 分别制备大鼠空白血清及四君子汤含药血清,MTT法检测IEC-6细胞增殖,RT-qPCR法检测p21、Cdk2 mRNA表达,Western blot法检测RhoA、p21、Cdk2及磷酸化Cdk2(p-Cdk2)蛋白表达。结果 与对照组比较,10%、20%四君子汤含药血清可促进给药24 h后细胞增殖(P<0.01),提高RhoA蛋白表达,Cdk2 mRNA和蛋白表达(P<0.05,P<0.01);各剂量四君子汤含药血清可促进给药48、72 h后细胞增殖(P<0.01),降低p21 mRNA和蛋白表达(P<0.01),升高p-Cdk2蛋白表达(P<0.05,P<0.01)。与DFMO组比较,各剂量四君子汤含药血清可促进给药48 h后细胞增殖(P<0.01),提高RhoA及p-Cdk2蛋白表达(P<0.01);10%、20%四君子汤含药血清可促进给药72 h后细胞增殖(P<0.01),降低p21 mRNA和蛋白表达(P<0.05,P<0.01),提高Cdk2 mRNA和蛋白表达(P<0.05,P<0.01)。结论 四君子汤含药血清可促进正常小肠上皮细胞增殖,其机制与影响多胺调控RhoA、p21、Cdk2表达有关。AIM To investigate the effects of Sijunzi Decoction-medicated serum on IEC-6 cell proliferation and expressions of Rho GTPases(RhoA),p21 and cyclin-dependent kinase 2(Cdk2).METHODS The SD rats serum with or without Sijunzi Decoction were both subjected to the detection of IEC-6 cell proliferation by MTT assay;the test of mRNA expressions of p21 and Cdk2 by RT-qPCR;the determination of protein expressions of RhoA,p21,Cdk2 and phospho-Cdk2(p-Cdk2)by Western blot.RESULTS Compared with the control group,the groups given 10%and 20%Sijunzi Decoction-medicated serum shared accelerated cell proliferation 24 h after administration(P<0.01),increased RhoA protein expression,and mRNA and protein expressions of Cdk2(P<0.05,P<0.01);all the groups given Sijunzi Decoction-medicated serum displayed promoted cell proliferation 48 h and 72 h after administration(P<0.01),decreased mRNA and protein expressions of p21(P<0.01)and improved p-Cdk2 protein expression(P<0.05,P<0.01).Compared with DFMO group,Sijunzi Decoction-medicated serum at all doses brought forth promoted cell proliferation 48 h after administration(P<0.01)and increased protein expressions of RhoA and p-Cdk2(P<0.01),which at 10%and 20%concentrations increased cell proliferation 72 h after administration(P<0.01),inhibited p21 mRNA and protein expressions and enhanced mRNA and protein expressions of Cdk2(P<0.05,P<0.01).CONCLUSION Sijunzi Decoction-medicated serum promote the proliferation of normal small intestinal epithelial cells,which may be related to the impact of its polyamines on the regulation of RhoA,p21 and Cdk2 expressions.

关 键 词：四君子汤 含药血清 IEC-6细胞 增殖 RHOA P21 CDK2 

分 类 号：R285.5[医药卫生—中药学]