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.
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)
出处
《中成药》
CAS
CSCD
北大核心
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)。