摘要
目的观察黄芪总皂苷对小肠上皮细胞(IEC-6)迁移和增殖的影响,并从多胺及其调控的细胞增殖相关蛋白(转录因子c-Myc、Rho-GTP酶RhoA和细胞周期依赖激酶Cdk2)表达角度研究其作用机制。方法从黄芪中提取黄芪总皂苷(皂苷含量88.8%)作为受试药,高效液相色谱仪蒸发光检测(HPLC-ELSD)获得受试药色谱图;移液器吸头划痕法检测细胞迁移率;MTT法检测细胞增殖;Western Blot法检测相关蛋白表达。结果黄芪总皂苷(25、50、100 mg·L^(-1))对细胞迁移无明显影响(与空白组比较,P>0.05)。黄芪总皂苷(50 mg·L^(-1)或100 mg·L^(-1))有促进细胞增殖作用(与空白组比较,P <0.05),且能逆转二氟甲基鸟氨酸(DFMO,多胺合成抑制剂)所致的细胞增殖抑制;可提高c-Myc、RhoA和Cdk2蛋白表达,逆转DFMO所致的c-Myc、RhoA和Cdk2蛋白表达抑制。结论黄芪总皂苷可促进IEC-6细胞增殖,其作用机制与影响多胺调控的增殖相关蛋白表达有关。
Objective This study aims to evaluate the effects of total saponins from Astragalus membranaceus on cellmigration and proliferation in intestinal epithelial cells(IEC-6),and to elucidate the potential mechanism throughpolyamines-mediated proliferation related proteins(transcription factor c-Myc,Rho GTPase RhoA and cell cycle-dependent kinase Cdk2). Methods The total saponins(88.8% purity) extracted from Astragalus membranaceus wasused as an administrated drug. High performance liquid chromatography with evaporative light detection(HPLC-ELSD) was performed to obtain the chromatogram of the administrated drug. Pipette tip scratch method was used todetect cell migration. MTT assay was carried out to determine cell proliferation. Western Blot analysis was used toevaluate the expression of proliferation related proteins. Results Treatment with total saponins from Astragalus membranaceus(25,50 and 100 mg·L^-1) had no effect on cell migration(P > 0.05,compared with blank group).However,total saponins from Astragalus membranaceus(50 mg·L^-1or 100 mg·L^-1) significantly promoted cellproliferation(P < 0.05,compared with the blank group),and reversed the inhibition of cell proliferation inducedby Difluoromethylornithine(DFMO, an inhibitor of polyamine synthesis). Total saponins from Astragalus membranaceus could increase the expression of c-Myc,RhoA and Cdk2 protein,and reverse the inhibition of c-Myc, RhoA and Cdk2 protein expression induced by DFMO. Conclusion Total saponins from Astragalus membranaceus promotes cell proliferation in IEC-6 partly through regulating the expression of polyamines-mediatedproliferation related proteins.
作者
朱易平
李茹柳
时玉霞
王东旭
梁雪丹
胡玲
陈蔚文
ZHU Yiping;LI Ruliu;SHI Yuxia;WANG Dongxu;LIANG Xuedan;HU Ling;CHEN Weiwen(Piwei Institute,Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China)
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2019年第1期60-65,共6页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
国家自然科学基金项目(81673940)
广州市科技计划项目(201607010335)
广州中医药大学"一流学科研究重点项目"(广中医规划[2018]6号)
关键词
黄芪总皂苷
小肠上皮细胞
细胞增殖
CDK2
RHOA
c-Myc
Total saponins from Astragalus membranaceus
intestinal epithelial cells
cell proliferation
Cdk2
RhoA
c-Myc
作者简介
朱易平,女,硕士研究生,研究方向:益气健脾方药作用机理及作用物质基础研究。Email:1054204652@qq.com;通信作者:李茹柳,博士研究生导师,教授,研究方向:益气健脾方药作用机理及作用物质基础研究。Email:lrl@gzuem.edu.cn。
