逍遥丸含药血清对人卵巢颗粒细胞内分泌功能的影响  

Effect of Xiaoyao Pill Containing Serum on the Endocrine Function of Human Ovarian Granulosa Cells

作　　者：杨宏敏[1] 宋翠淼[2] 周楠[1] 陆君[1] 路帅[1] 马惠荣[1] 段彦苍[1] 徐华洲[2] 杜惠兰[1] 

机构地区：[1]河北医科大学中西医结合学院 [2]河北医科大学基础课教学部

出　　处：《中医杂志》2013年第21期1850-1853,1866,共5页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(30772818)

摘　　要：目的探讨逍遥丸对卵巢颗粒细胞内分泌功能影响可能的分子机制。方法 12只Wistar大鼠随机分为逍遥丸低、中、高剂量组(每日灌服逍遥丸混悬液1ml/100g、2ml/100g、3ml/100g)和对照组(每日灌服生理盐水2ml/100g),连续4天,于末次灌胃后1h大鼠股动脉采血制备低、中、高剂量逍遥丸大鼠含药血清及正常大鼠血清。收集行体外受精-胚胎移植患者卵泡液,提取颗粒细胞原代细胞培养24h,以低、中、高剂量逍遥丸大鼠含药血清以及正常大鼠血清分别与体外培养的人卵巢颗粒细胞共同孵育48h,分为逍遥丸低、中、高剂量组以及对照组。采用放射免疫方法测定培养液中雌二醇(E2)、孕酮(P)、环磷酸腺苷(cAMP)的水平,采用蛋白印迹法和实时定量荧光PCR法分别检测促卵泡素受体(FSHR)蛋白和FSHR mRNA、P450芳香化酶mRNA、过氧化物酶体增殖物激活受体-γ(PPARγ)mRNA在卵巢颗粒细胞的表达。结果与对照组比较,逍遥丸中、高剂量组培养液中E2、cAMP水平明显升高,逍遥丸低、中、高剂量组FSHR蛋白表达明显升高,逍遥丸高剂量组FSHR、P450芳香化酶mRNA表达明显升高,逍遥丸中剂量组PPAR γmRNA表达明显降低(P<0.05或P<0.01)。与逍遥丸低剂量组比较,逍遥丸高剂量组E2、cAMP、FSHR蛋白及mRNA水平明显升高(P<0.01)。结论逍遥丸可能通过增加卵巢颗粒细胞FSHR蛋白及其mRNA的表达,使FSH更好发挥促进卵泡发育、排卵的作用,并通过调节FSHR、cAMP、P450芳香化酶、E2水平促进卵巢颗粒细胞的内分泌功能。Objective To research the possible molecular mechanism of Xiaoyao Pill （Free Wanderer Pill） containing serum on the endocrine function of ovarian granulose cells. Methods Twelve Wistar rats were randomized into Xiaoyao Pill low-dose, middle-dose and high-dose groups （1 ml/100 g, 2 ml/100 g and 3 ml/100 g Xiaoyao Pill suspension was garaged to each rat daily） and control group （each rat was gavaged with 2 ml/100 g normal saline daily）. The administration lasted for 4 days. Xiaoyao Pill low-dose, middle-dose and high-dose rat containing serum and normal rat serum was prepared by collecting blood from the femoral artery lh after the last administration. The follicular fluid of in vitro fertilization and embryo transfer （IVF ET） patients was collected to extract granule cells and culture primary cells for 24 hours. Xiaoyao Pill low-dose, middle-dose and high-dose rat containing serum and normal rat serum was incubated with cultured human ovarian granulosa cells together for 48 hours respectively and divided into Xiaoyao Pill low-dose, middle-dose and high-dose groups and control group. The levels of estradiol （E2）, progesterone （P） and cyclic adenosine monophosphate （cAMP） in culture solution were detected with radioimmunoassay. The Western blot and Real-time PCR were used to detect the expressions of follicle stimulating hormone receptor （FSHR） protein, FSHR mRNA, P450 aromatase mRNA and peroxisome proliferator-activated receptor-γ （PPARγ） mRNA in ovarian granulosa cells. Results Comparing with the control group, the levels of E2 and cAMP in Xiaoyao Pill middle-dose and high-dose groups were significantly increased, the FSHR protein expression in Xiaoyao Pill low-dose, middle-dose and high-dose groups was significantly increased, the expressions of FSHR mRNA and P450 aromatase mRNA in Xiaoyao Pill high-dose group were significantly increased and the PPARγ mRNA expression in Xiaoyao Pill middle-dose group was significantly decreased （P〈0.05 or P〈0.01）. Comparing with Xia

关 键 词：逍遥丸 颗粒细胞 内分泌 卵泡刺激素受体 卵巢功能失调 

分 类 号：R271.9[医药卫生—中西医结合] R285.5[医药卫生—中医妇科学]