滋阴补阳序贯法联合西药对DOR大鼠卵巢Smad2、Smad3、Smad7 mRNA表达的影响

Effect of Nourishing Yin and Nonifying Yang Sequential Therapy Combined with Western Medicine on Expression Levels of Smad2,Smad3,Smad7 mRNA in Rats with Diminished Ovarian Reserve

目的探讨滋阴补阳序贯法联合西药对卵巢储备功能下降(DOR)大鼠卵巢Smad2、Smad3、Smad7 mRNA表达的影响。方法将40只SD雌性大鼠随机分为空白组、模型组、西药组、滋阴补阳序贯组、联合用药组(西药+滋阴补阳序贯中药),每组8只。采用雷公藤多苷片50mg/(kg·d)灌胃连续2周,建立DOR大鼠模型。其后空白组、模型组给予生理盐水灌胃,连续10d;西药组予人工周期加促排卵治疗;滋阴补阳序贯组在动情前期予滋阴方、动情后期予补阳方灌胃(4.32g/kg);联合用药组在西药组的基础上动情前期予滋阴方,动情后期予补阳方。10d后用ELISA法检测大鼠血清卵泡刺激素(FSH)、黄体生成素(LH)、雌二醇(E2)、抗苗勒管激素(AMH)、转化生长因子-β1(TGF-β1)、抑制素B(INHB)水平,Real-time PCR法检测Smad2、Smad3、Smad7 mRNA的相对含量。结果与模型组比较,西药组、滋阴补阳序贯组和联合用药组FSH及LH水平降低(均P<0.01),联合用药组低于西药组(均P<0.05);各用药组大鼠血清E2、AMH、TGF-β1、INHB水平明显升高(P<0.05,P<0.01),联合用药组高于西药组(P<0.01,P<0.05)。与模型组比较,滋阴补阳序贯组及联合用药组Smad2 mRNA的表达升高,联合用药组Smad3 mRNA的表达升高(均P<0.01),且联合用药组Smad2、Smad3 mRNA均高于西药组(均P<0.01),各用药组Smad7 mRNA表达均下降(均P<0.01)。结论滋阴补阳序贯法联合西药能上调DOR大鼠卵巢Smad2、Smad3 mRNA表达,下调Smad7 mRNA的表达,改善卵巢功能。

Objective To investigate the effect of Nourishing Yin and Nonifying Yang sequential therapy（NYNYST）combined with western medicine on expression levels of Smad2,Smad3,Smad7 mRNA in rats with diminished ovarian reserve（DOR）.Methods Totally 40 female rats were randomly divided into 5 groups,the normal control group,the model group,the western medicine group,the NYNYST group and the combination group（western medicine combined with NYNYST）,8 in each group.The DOR model was established through orally administering tripterygium pill for continuous 2 weeks.The normal control group and the model group were administered with saline for 10 days.The western medicine group was treated with hormone replacement therapy（HRT）and ovarian stimulation.The NYNYST group was administered with Nourishing Yin herbs in proestrus and Nonifying Yang herbs in late estrus and the combination group was administered with combination of Chinese herb and western drugs for 10 days,with the same dose in the former two groups.Serum levels of FSH,LH,E2,anti-Müllerian hormone（AMH）,Transforming growth factor beta 1（TGF-β1）and Inhibin B（INHB）were measured by ELISA.Changes of Smad2,Smad3,Smad7 mRNA in ovaries were detected by real-time PCR.Results Compared with the model group,the serum levels of FSH,LH were decreased significantly in western medicine group,NYNYST group and combination group（P〈0.01）,the serum levels of E2,AMH,TGF-β1 and INHB increased in the rats of the treatment group（P〈0.05,P〈0.01）,and efficacy of the combination group was significantly superior to that of the western medicine group（P〈0.01,P〈0.05）;compared with the model group,Smad2 mRNA increased significantly in NYNYST group and combination group,Smad3 mRNA increased sig-nificantly in combination group（P〈0.01）,the efficacy of combination group was superior to that of the western medicine group（P〈0.05）;compared with the model group,Smad7 mRNA of treatment groups was decreased significantly（P〈0.01）.Conclusion NYTYST combined with western medicine can improve the function of ovaries by up-regulating the expression of Smad2,Smad3 mRNA and down-regulating the expression of Smad7 mRNA in DOR rats.