摘要
目的研究“补气生津”人参-黄芪与“填精补肾”熟地-山茱萸对皮质激素生成的影响。方法采用0.25-8 g·L^(-1)人参-黄芪与0.25-8 g·L^(-1)熟地-山茱萸水煎醇沉液分别干预小鼠肾上腺皮质瘤细胞(Y1 adrenocortical cell,Y1细胞)48 h,采用MTT比色法(MTT assay,MTT)检测细胞增殖,运用酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测细胞分泌皮质酮含量,实时荧光定量PCR(Real-time Quantitative polymerase chain reaction,qPCR)检测类固醇激素合成酶基因表达,免疫蛋白印迹法(Western blot,WB)检测蛋白表达。结果与空白组(Control,Con)比较,0.25-8 g·L^(-1)人参-黄芪和熟地-山茱萸均抑制Y1细胞增殖(P<0.01);4 g·L^(-1)和8 g·L^(-1)人参-黄芪促进皮质酮分泌(P<0.01),4 g·L^(-1)和8 g·L^(-1)熟地-山茱萸抑制皮质酮分泌(P<0.01);8 g·L^(-1)人参-黄芪增强CYP11A1、CYP21A2、CYP11B1蛋白表达(P<0.05),8 g·L^(-1)熟地-山茱萸抑制CYP11A1、CYP21A2、CYP11B1蛋白表达(P<0.01);8 g·L^(-1)人参-黄芪抑制Star基因表达(P<0.05),而8 g·L^(-1)熟地-山茱萸促进Star基因表达(P<0.01),4 g·L^(-1)与8 g·L^(-1)人参-黄芪和0.25-8 g·L^(-1)熟地-山茱萸均能显著抑制Cyp11a1基因表达(P<0.01),8 g·L^(-1)人参-黄芪和1-8 g·L^(-1)熟地-山茱萸都抑制Cyp21a1基因表达(P<0.05),1-8 g·L^(-1)人参-黄芪和0.25-8 g·L^(-1)熟地-山茱萸均抑制了Cyp11b1基因表达(P<0.05)。与佛司可林(Forskolin,FSK)组比较,人参-黄芪与熟地-山茱萸预处理Y1细胞后,FSK不能诱导Star、Cyp11a1、Cyp21a1、Cyp11b11基因表达(P<0.01)。结论人参-黄芪促进皮质酮分泌,熟地-山茱萸抑制皮质酮分泌,主要通过调节类固醇激素合成酶表达发挥药效。
Objective To study the effects of“Nourish Qi and Produce Fluid”Radix Ginseng-Radix Astragali Seu Hedysari and“Replenish essence and kidney”Rehmanniae Radix Praeparata-Fructus Corni on corticosterone secretion.Methods Y1 mouse adrenocortical tumor cells were treated with 0.05-8 g·L^(-1)total extract of Radix Ginseng-Radix Astragali Seu Hedysari and 0.25-8 g·L^(-1)Rehmanniae Radix Praeparata-Fructus Corni for 48 h.Then,Y1 cells proliferation were tested with MTT method.Corticosterone were tested by ELISA(Enzyme Linked Immunosorbent Assay).mRNA expression was detected by Real time quantitative PCR(Polymerase Chain Reaction)and protein expression were carried out by WB(Western blot).Results Compared with the control group,the proliferation of Y1 cells was inhibited by 0.25-8 g·L^(-1)Radix Ginseng-Radix Astragali Seu Hedysari and Rehmanniae Radix PraeparataFructus Corni(P<0.01);4 g·L^(-1)and 8g·L^(-1)Radix Ginseng-Radix Astragali Seu Hedysari promotes corticosterone secretion(P<0.01);The protein expression of CYP11A1,CYP21A2,CYP11B1(P<0.05)was enhanced by 8 g·L^(-1)Radix Ginseng-Radix Astragali Seu Hedysari,and the Rehmanniae Radix Praeparata-Fructus Corni inhibited the protein expression of CYP11A1,CYP21A2,and CYP11B1(P<0.01);Star gene expression was inhibited by 8 g·L^(-1)Radix Ginseng-Radix Astragali Seu Hedysari(P<0.05),while 8 g·L^(-1)Rehmanniae Radix Praeparata-Fructus Corni promoted Star gene expression(P<0.01).Cyp11a1 gene expression was inhibited by 4 g·L^(-1)and 8 g·L^(-1)Radix Ginseng-Radix Astragali Seu Hedysari,0.25-8 g·L^(-1)Rehmanniae Radix Praeparata-Fructus Corni(P<0.01);Cyp21a1 gene expression was inhibited by 8 g·L^(-1)Radix Ginseng-Radix Astragali Seu Hedysari and 1-8 g·L^(-1)Rehmanniae Radix PraeparataFructus Corni(P<0.05);Cyp11b1 gene expression was inhibited by1-8 g·L^(-1)Radix Ginseng-Radix Astragali Seu Hedysari 0.25-8 g·L^(-1)Rehmanniae Radix Praeparata-Fructus Corni(P<0.05).Compared with theForskolin(FSK)group,after pretreatment of Y1 cells with Radix Ginseng-Radix Astragali Seu Hedysari and Rehmanniae Radix Praeparata-Fructus Corni the gene expression of Star,Cyp11a1,Cyp21a1,and Cyp11b11 could not be induced by FSK(P<0.01).Conclusion Radix Ginseng-Radix Astragali Seu Hedysari promote the secretion of corticosterone,and Rehmanniae Radix Praeparata-Fructus Corni inhibits the secretion of corticosterone,mainly by regulating the expression of steroid hormone synthase.
作者
谢海纳
潘志强
Xie Haina;Pan Zhiqiang(Basic Medical School,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2021年第10期3638-3645,共8页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
上海市科学技术委员会科技创新行动计划(19140905000):糖皮质激素诱发药源性脾肾虚证小鼠模型的创建及其指标评价体系研究,负责人:潘志强。
关键词
人参-黄芪
熟地-山茱萸
中药药对
肾上腺皮质
皮质酮
Y1细胞
Radix Ginseng-Radix Astragali Seu Hedysari
Rehmanniae Radix Praeparata-Fructus Corni
Chinese medicine couplet medicinals
Adrenocortical
Corticosterone
Y1 adrenocortical cell
