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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 effects of Bushen Tiaojing Recipe and Xiaoyao Pill on the expression of GDF-9, BMP-15, Protein and its mRNA expression, analysis of kidney method, Shugan method induced ovulation possible molecular mechanism. Methods The immature female mice were randomly divided into control group, Bushen group and Shugan group. All mice were injected intraperitoneally with 5 IU / kg of PMSG on the third day, h CG), 5 IU / kg, kidney-tonifying group and liver-clearing group were administrated Bushen Tiaojing Decoction respectively (the first and third days were fed with the second dose of 18.70 g / kg, the second and fifth days were fed with the third dose of 28.57 g / kg) and Xiaoyao Pill (the high dose of 2.34 g / kg on days 1 to 3 and the low dose of 1.17 g / kg on days 4 to 5) to promote follicular development and induce ovulation, real-time quantitative PCR, Western blot, The expression of GDF-9, BMP-15 protein and mRNA in ovary at 0, 8 and 12 h after h CG injection were analyzed by immunohistochemistry. Results Compared with the control group, Bushen Tiaojing Recipe and Xiaoyao Pill could increase the expression of GDF-9 and BMP-15 mRNA and protein in ovulation induced mice 8 h and 12 h after h CG injection (all P <0.05) . Conclusion Bushen Tiaojing Recipe and Xiaoyao Pill can promote the proliferation of follicular granulosa cells, the proliferation of cumulus follicles and the maturation of oocytes by the effect on ovarian oocyte secretion factors GDF-9 and BMP-15 protein and their mRNA expression before ovulation in ovulation induction mice , And then play a role in promoting ovulation induction.