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目的在前期研究的基础上,探讨以“补肾调冲法”立法的通脉大生片对大鼠卵巢TGF-β/Smads信号转导通路的调控作用,探索中医补肾调冲法促卵泡发育的机制,从而为补肾调冲法治疗肾虚排卵障碍型不孕提供科学依据。方法选取9日龄SD大鼠,通过给予颈背部皮下注射丙酸睾酮,建立肾虚排卵障碍型不孕大鼠(ASR)模型,根据大鼠阴道脱落细胞涂片的表现,筛选出造模成功的大鼠,随机分为6个组,分别为模型组、通脉大生片高剂量组(简称通高组)、通脉大生片中剂量组(简称通中组)、通脉大生片低剂量组(简称通低组)、来曲唑组(LE)、右归丸组,另设正常组,于大鼠第80日龄,分别给予相应药物进行干预,连续干预5周后股动脉取血并处死动物,迅速剥离大鼠卵巢,每组选取三只大鼠的卵巢组织,立即置于Eppendorf管中,储存于液氮中以备检测。采用免疫印迹法,检测各组大鼠卵巢P-Smad3的表达水平。结果与模型组相比,正常组、通高组、来曲唑组、右归丸组大鼠卵巢P-Smad3的表达水平较高,差异有显著意义(P<0.01),通低组、通中组的P-Smad3水平亦有所升高,差异有统计学意义(P<0.05);通脉大生片各剂量组中,通高组升高P-Smad3水平的效果明显高于通中组、通低组。结论通脉大生片可以提高肾虚排卵障碍性不孕大鼠P-smad3的表达水平,进而改善卵巢局部的微环境,以促进卵泡的生长、发育和促进成熟卵泡破裂及卵子的排出,从而达到治疗肾虚排卵障碍性不孕的作用。
OBJECTIVE: To investigate the regulation of TGF-β / Smads signaling pathway in rat ovary by Tongmaishengchuan tablet with “Bushen tiaochong law” legislation based on the previous study, So as to provide a scientific basis for the treatment of kidney deficiency and ovulation disorder infertility by regulating kidney and regulating blood flow. Methods 9-day-old SD rats were selected and injected with testosterone propionate subcutaneously on the back of the neck to establish a rat model of infertility induced by renal dysplasia (ASR). According to the appearance of vaginal smear in rats, Rats were randomly divided into 6 groups: model group, Tongmaishengsheng high dose group (Tonggao group), Tongmaishengsheng medium dose group (Tongzhong group), Tongmaishengsheng low dose group (LE), Yougui Wan group, another normal group, at the age of 80 days, were given the appropriate drug intervention, continuous intervention 5 weeks after the femoral artery blood and The animals were sacrificed and the ovaries of rats were rapidly dissected. The ovaries of three rats in each group were selected and immediately placed in Eppendorf tubes and stored in liquid nitrogen for testing. Immunohistochemistry was used to detect the expression of P-Smad3 in ovary of each group. Results Compared with model group, the expression of P-Smad3 in ovary of normal group, Tong Gao group, Letrozole group and Yougui Wan group were higher than that in model group (P <0.01) The P-Smad3 level in the middle group was also increased, the difference was statistically significant (P <0.05); Tongmaishengsheng tablet dose group, Tong Gao group increased P-Smad3 levels were significantly higher than the Tongzhong group , Pass low group. Conclusions Tongmaishengsheng Tablet can improve the expression of P-smad3 in rats with renal dysfunction of ovulation disorder, and then improve the micro-environment in ovary to promote the growth and development of follicles and promote the rupture of mature follicles and the release of eggs, so as to achieve the treatment Kidney anovulatory infertility role.