黄芪多糖离体条件下对H295R细胞维生素D系统相关蛋白表达及睾酮分泌的影响  

Effect of Astragalus Polysaccharide on Protein Expression of Vitamin D System and Secretion of Testosterone in H295R Cells in Vitro

作　　者：杨凡 管连城 柴艺汇 高洁 李文 秦忠 陈云志 Yang Fan;Guan Liancheng;Chai Yihui;Gao Jie;Li Wen;Qin Zhong;Chen Yunzhi(Second Hospital Affiliated to Guizhou University of TCM,Guiyang 550003;chool of Basic Medical Sciences,Guizhou University of TCM,Guiyang 550002;School of Basic Medical Sciences,Guizhou University of TCM,Guiyang 550002)

机构地区：[1]贵州中医药大学第二附属医院,贵阳550002 [2]贵州中医药大学实验中心,贵阳550025 [3]贵州中医药大学基础医学院,贵阳550025

出　　处：《中药药理与临床》2020年第2期132-136,共5页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金项目(编号:81874376、81373517、81660762);贵州省科技计划项目[黔科合基础(2017)1401];贵州省教育厅创新群体重大研究项目[黔教合KY字(2017)041];黔科合平台人才[(2018)5766号-5]。

摘　　要：目的:探讨黄芪多糖对人肾上腺皮质细胞(H295R细胞)维生素D系统及睾酮表达的影响。方法:随机分为空白对照组、维生素D组、黄芪多糖25μg/mL、50μg/mL、100μg/mL组,四唑盐(MTT)法检测不同浓度的黄芪多糖和维生素D对H295R细胞增殖率的影响,ELISA法检测睾酮分泌水平,实时荧光定量PCR(RT-qPCR)检测维生素D受体(VDR)、25-羟基维生素D3-1α-羟化酶(CYP27B)、维生素D3-24-羟化酶(CYP24A)、17β-羟类固醇脱氢酶(17β-HSD)、19α-羟化酶(CYP19A1)mRNA表达,Western blot法检测VDR、CYP27B、CYP24A蛋白表达。结果:黄芪多糖12.5μg/mL浓度,维生素D在0.25×10^-11 mol/L和0.5×10^-11 mol/L浓度,对H295R细胞增殖无显著影响。黄芪多糖200μg/mL,维生素D在2×10^-11 mol/L和4×10^-11 mol/L浓度时对H295R细胞增殖存在一定抑制作用。与空白对照组比较,维生素D组与黄芪多糖100μg/mL组睾酮分泌水平显著升高,VDR、CYP24A、17β-HSD mRNA表达显著上调,CYP27B蛋白及mRNA、CYP19A1 mRNA表达明显下调(P<0.05或P<0.01)。结论:离体条件下,黄芪多糖可能通过调节H295R细胞维生素D系统促进睾酮合成分泌。Objective: To investigate the effects of astragalus polysaccharides on vitamin D system and testosterone expression in human adrenal cortex cells(H295 R cells). Methods: H295 R cells were randomly divided into the control group, vitamin D group, astragalus polysaccharide 25 μg/ml, 50 μg/ml, 100 μg/ml groups. MTT method was used to detect the proliferation effect on H295 R cells of astragalus polysaccharides and vitamin D in different concentrations. The secretion level of testosterone was detected by ELISA. The mRNA expressions of VDR, CYP27 B, CYP24 A, 17β-HSD and CYP19 A1 were detected by RT-qPCR. The protein expressions of VDR, CYP27 B and CYP24 A were detected by Western blot. Results:No significant effect on the proliferation of H295 R cells was observed in Astragalus polysaccharide 12.5 μg/ml, vitamin D 0.25×10^-11 mol/L and 0.5×10^-11 mol/l groups. Certain inhibitory effect was observed in Astragalus polysaccharide 200 μg/ml, vitamin D 2×10^-11 mol/L group and 4×10^-11 mol/l group. Compared with the control group, the levels of testosterone secretion were significantly increased in the vitamin D group and astragalus polysaccharides 100 g/mL group, the mRNA expressions of VDR, CYP24 A and 17β-HSD were significantly up-regulated, the mRNA expressions of CYP27 B and CYP19 A1 were significantly down-regulated(P<0.05). Conclusion: Astragalus polysaccharides can promote the synthesis and secretion of testosterone by regulating the vitamin D system in H295 R cells.

关 键 词：黄芪多糖 H295R细胞 维生素D系统 睾酮 

分 类 号：R285[医药卫生—中药学]