紫草多糖对培养大鼠颗粒细胞生成甾体激素的影响

Effect of Arnebia Euchroma Glycosaminoglycans on Steroidogenesis of Cultured Granulosa Cells from Rat in vitro

目的研究中药紫草的水溶成分——紫草多糖对体外培养颗粒细胞的直接作用。方法分离出大鼠双侧卵巢的颗粒细胞进行体外培养。选取健康未成年雌性SD大鼠160只,其中60只按随机数字表法分为基础组和hCG组,分别采用不同浓度(0.00、0.30、0.75、1.50、3.00、7.50 g.L-1)紫草多糖培养,每组5只;另100只按随机数字表法分为空白对照组、紫草多糖组、hCG组和hCG-紫草多糖组,分别进行不同时间(0、1、2、3、4 h)的培养,每组5只,共计100只。采用放射免疫测定法(RIA)测定不同实验组颗粒细胞培养液中孕酮(Progesterone,P4)和雌二醇(Estradiol,E2)的浓度。结果紫草多糖呈剂量依赖性地抑制hCG刺激下颗粒细胞P4和E2的生成,大剂量(大于3.00 g.L-1)也抑制基础P4和E2的生成,紫草多糖主要使培养液中的P4含量降低,但大剂量也可使细胞内的P4含量降低。紫草多糖作用于颗粒细胞1 h后明显抑制hCG促P4生成的作用,但细胞洗涤后再培养,对hCG的反应性与对照组比较无明显变化。结论紫草多糖可直接作用于体外培养的颗粒细胞,抑制hCG促甾体激素生成的作用,该抑制作用起效时间短,并且是可逆的。

Objective To study the effect of soluble component-Arnebia Euchroma Glycosaminoglycans（AEG） on steroidogenesis of cultured granulosa cells from rat ovary in vitro.Methods Ovarian granulosa cells were isolated and cultured in vitro.Total 160 unmatured female SD rats were selected.60 rats were randomly divided into basic groups and hCG-treated groups,the granulosa cells of these groups were cultured with different concentrations of AEG（0.00,0.30,0.75,1.50,3.00,7.50 g·L^-1）,5 in each group.Then 100 rats were randomly divided into blank control groups,AEG groups,hCG-treated groups,hCG and AEG treated groups,the granulosa cells of these groups were cultured with different time（0,1,2,3,4 h）,5 in each group.Use the method of RIA to evaluate concentrations of P4 and E2 in different experimental groups.Results AEG could inhibit P4 and E2 productions of hCG-treated granulosa cells in dose-dependent manners,and large dosages（〉3.00 g·L^-1） could inhibit basic P4 and E2 productions.AEG could reduce the content of P4 in culture fluid,but large dosages could reduce the content of P4 in cells.After granulosa cells were treated with AEG for 1 hour,the effect of hCG-induced P4 production was obviously inhibited,but compared with control group,the response of cells to hCG had not significant difference by washed these cells.Conclusion AEG could affect granulosa cells cultured in vitro to inhibit hCG-induced steroidogenesis.The inhibition had a rapid onset,and it was reversible.