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补肾调经方、逍遥丸对促排卵小鼠排卵前卵巢卵母细胞分泌因子表达的影响
Effects of Bushen Tiaojing Fang and Xiaoyao Wan on the expression of oocyte secreted factors in mouse ovary before ovulation

作  者: (段彦苍); (宋翠淼); (贺明); (杜惠兰); (刘亚华);

机构地区: 河北中医学院中西医结合学院,河北050200

出  处: 《北京中医药大学学报》 2017年第8期635-640,共6页

摘  要: 目的探讨补肾调经方、逍遥丸对促排卵小鼠排卵前卵巢中影响排卵率的重要因子卵母细胞分泌因子生长分化因子-9(GDF-9)、骨形态发生蛋白-15(BMP-15)蛋白及其mRNA表达的影响,分析补肾法、疏肝法诱发排卵可能的分子机制。方法将性未成熟雌性小鼠随机分为对照组、补肾组和疏肝组,均于第3天腹腔注射血促性素(PMSG)5 IU/只、第5天腹腔注射绒促性素(h CG)5 IU/只,补肾组、疏肝组分别灌服补肾调经方(其中第1~3天灌服Ⅱ号方18.70 g/kg,第4~5天灌服Ⅲ号方28.57 g/kg)和逍遥丸(其中第1~3天灌服高剂量2.34 g/kg,第4~5天灌服低剂量1.17 g/kg)促进卵泡发育并诱发排卵,实时荧光定量PCR、Western blot、免疫组化分析注射h CG后0、8、12h卵巢GDF-9、BMP-15蛋白及其mRNA的表达。结果与对照组比较,补肾调经方、逍遥丸均可使促排卵小鼠在注射h CG后8、12 h GDF-9、BMP-15 mRNA及其蛋白的表达均升高(均P<0.05)。结论补肾调经方、逍遥丸通过对促排卵小鼠排卵前卵巢卵母细胞分泌因子GDF-9、BMP-15蛋白及其mRNA表达的影响,促进卵泡颗粒细胞增殖、卵丘扩散、卵母细胞成熟,进而发挥促进诱发排卵的作用。 Objective To investigate the possible mechanisms of ovulation induced by supplementing kidney and smoothing liver by observing the effects of Bushen Tiaojing Fang( Kidney-tonifying Menstrua- tion-regulating Formula, BSTJF) and Xiaoyao Wan (Free Wanderer Pill, XYW) on the expression of growth differentiation factor-9 ( GDF-9 ), bone morphogenetic protein ( BMP-15 ) mRNA and its protein expression. Methods Immature female mice were randomly divided into normal control group, kidney- tonifying(BSTJF) group and liver-soothing(XYW) group. All groups received intraperitoneal injection of 5 IU pregnant mare serum gonadotrophin(PMSG) at day 3 and 5 IU human chorionic gonadotropin(hCG) at day 5 ; mice in kidney-tonifying group and liver-soothing group received oral gavage of BSTJF ( 18.70 g/kg BSTJF II from day 1 to day 3 ; 28.57 g/kg BSTJF III from day 4 to day 5 ) or XYW (2.34 g/kgXYW from day 1 to day 3 ; 1.17 g/kg XYW from day 4 to day 5) respectively. GDF-9, BMP-15mRNA and its protein expression in ovary were measured with real-time fluorescence quantitative PCR, Western blot, and immunohistochemical analysis at the 0,8,12 hours after hCG injection. Results Compared with the normal control group, BSTJF and XYW increased the expressions of GDF-9, BMP-15mRNA and its protein at 8 and 12 hours after hCG injection(P 〈0. 05). Conclusion Bushen Tiaofing Fang and Xiaoyao Wan may induce ovulation by promoting proliferation of ovarian granular ceils, expansion of cu- mulus oophorus, and maturity of oocyte through increasing the expression of GDF-9, BMP-15mRNA and its protein.

关 键 词: 补肾调经方 逍遥丸 排卵 小鼠

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