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目的观察左归丸对免疫性卵巢早衰(premature ovarian failure,POF)模型小鼠卵巢生长分化因子-9(growth differentiation factor-9,GDF-9)/Smad2信号转导的影响,探讨左归丸治疗POF的机制。方法将84只雌性Balb/c小鼠随机分为空白对照组(12只)、模型组(24只)、左归丸组(24只)及己烯雌酚组(24只)。除空白对照组外,其余3组以小鼠透明带多肽3(zona pellucida 3,ZP3)为抗原,皮下多点注射建立POF模型,左归丸组及己烯雌酚组分别给予2.7g/(kg·d)左归丸及0.15mg/(kg·d)己烯雌酚水溶液灌胃,每日1次,每周6次,连续灌胃4、8周。观察各组小鼠卵巢生长卵泡比率;分别采用免疫组化法及Western blot法检测小鼠卵巢组织GDF-9及Smad2蛋白表达水平。结果与空白对照组比较,模型组造模4、8周小鼠卵巢生长卵泡比率及GDF-9、Smad2蛋白表达均明显减少(P<0.05);与模型组同期比较,左归丸及己烯雌酚组小鼠卵巢生长卵泡比率、GDF-9蛋白水平表达明显升高(P<0.05),左归丸组8周Smad2蛋白水平表达升高(P<0.05)。结论POF的发病可能与卵巢GDF-9蛋白表达水平不足,GDF-9/Smad2信号转导功能低下,导致始基卵泡不能有效募集,原位闭锁有关。左归丸能增加卵泡GDF-9蛋白表达,其机制可能是通过GDF-9/Smad2信号转导通路对卵泡生长发育进行调节,从而发挥治疗POF的作用。
Objective To observe the effect of Zuogui pill on the signal transduction of ovary growth differentiation factor-9 (GDF-9) / Smad2 in the mouse model of premature ovarian failure (POF) POF mechanism. Methods Totally 84 female Balb / c mice were randomly divided into blank control group (n = 12), model group (n = 24), Zuogui pill group (24) and diethylstilbestrol group (n = 24). Except for the blank control group, POF model was established by subcutaneously injection of Zona pellucida 3 (ZP3) in the remaining three groups. The mice in Zuogui pill group and diethylstilbestrol group were given 2.7g / (kg · d ) Zuogui Pills and diethylstilbestrol 0.15mg / (kg · d) aqueous solution, once a day, 6 times a week, continuous gavage 4,8 weeks. The ratio of ovarian follicles in each group was observed. The protein expression of GDF-9 and Smad2 in ovary tissue of mice was detected by immunohistochemistry and Western blot respectively. Results Compared with the blank control group, the ratio of ovarian follicle growth and the expression of GDF-9 and Smad2 in model group were significantly decreased at 4 and 8 weeks (P <0.05). Compared with the model group, Zuogui pill and diethylstilbestrol group The ovarian growth follicle ratio and GDF-9 protein level in mice increased significantly (P <0.05), and the expression of Smad2 protein in Zuoguiwan group increased 8 weeks (P <0.05). Conclusions The onset of POF may be related to the lack of expression of GDF-9 protein in ovary and the low signal transduction function of GDF-9 / Smad2, resulting in the inability of primordial follicles to recruit. Zuogui pill can increase the expression of GDF-9 protein in follicles, and its mechanism may be through the GDF-9 / Smad2 signaling pathway on follicular growth and development regulation, thus play a role in the treatment of POF.