补肾法与疏肝法对小鼠卵母细胞质量及卵母细胞骨形态发生蛋白-15表达的影响

Effects of Bushen treatment and Shugan treatment on oocyte mass and bone morphogenetic protein-15 of mice

目的:研究补肾法、疏肝法提高控制性超排卵小鼠卵母细胞质量的作用机制。方法:将300只雌性昆明小鼠随机分为空白对照组,控制性超排卵组(COH组),疏肝低、高剂量组,补肾低、高剂量组,每组50只。按照1mL/100g体质量灌胃10d,空白对照组和COH组给予蒸馏水灌胃,疏肝低、高剂量组分别给予逍遥丸0.3g/mL、0.6g/mL灌胃,补肾低、高剂量组分别给予补肾调经方2.7g/mL、5.4g/mL灌胃。第11天时除空白对照组外其余小鼠腹腔注射PMSG 10IU,48h后腹腔注射HCG 10IU,16h后进行处死。检测各组排卵数及优质卵泡率;实时荧光定量PCR法检测各组小鼠卵母细胞骨形态发生蛋白-15(BMP-15)mRNA表达;免疫组化和Western blot检测各组小鼠卵母细胞BMP-15蛋白表达。结果:各治疗组及COH组平均排卵数显著高于空白对照组(P<0.05);各治疗组优质卵泡率显著高于COH组(P<0.05);各治疗组卵母细胞BMP-15 mRNA及蛋白表达显著高于COH组(P<0.05)。结论:补肾法、疏肝法可提高控制性超排卵小鼠的排卵数和优质卵泡率,其作用机制与调控卵母细胞BMP-15的表达有关。

Objective:To study the effects of Bushen treatment and Shugan treatment on bone morphogenetic protein-15(BMP-15)in superovulated mice.Methods:A total of 300 mice were divided randomly:control group,COH group,Shugan low dose group,Shugan high dose group,Bushen low dose group,Bushen high dose group,50 mice in each group.The control group and the COH group were drenched with distilled water,Shugan low and high dose group were drenched with 0.3g/mL and 0.6g/mL refined oral solution of Xiaoyao Pills,Bushen low and high dose group were drenched with 2.7g/mL and 5.4g/mL refined oral solution of Bushen Tiaojing Recipe,1mL/100g weight for 10 days consecutively.On the eleventh day,the treatment groups and the COH group were dealt with intraperitoneal injection of PMSG 10IU,then intraperitoneal injection of HCG 10IU after 48h.The mice were killed after 16h.The follicles number and the high-quality follicle rate were detected.The mRNA expressions of BMP-15 in oocytes were detected by real-time fluorescent quantitative PCR.The protein expressions of BMP-15 in oocytes were detected by immunohistochemistry and Western blot.Results:The follicles number of the COH group and the treatment groups were increased significantly than the control group(P<0.05).The high-quality follicle rates of the treatment groups were increased significantly than the COH group(P<0.05).The BMP-15 expressions of the treatment groups were increased significantly than the COH group(P<0.05).Conclusion:Shugan treatment and Bushen treatment can improve the follicles number and the high-quality follicle rate of the mice,which may be relevant to up-regulating the expression of BMP-15 in oocytes.