金氏补肾活血方对卵巢储备功能减退大鼠TGF-β1/Smads信号通路及性激素的影响

Effect of Jin’s Bushen Huoxue Decoction on TGF-β1/Smads signal pathway and sex hormones in rats with ovarian reserve dysfunction

目的观察金氏补肾活血方对卵巢储备功能减退模型大鼠的性激素、卵巢组织形态的作用及其调节转化生长因子-β1(TGF-β1)/Smads信号通路的可能机制。方法将50只SD雌性大鼠随机分为5组:空白组、模型组、西药组、中药低剂量组、中药高剂量组,每组10只。除空白组外,其余各组均给予环磷酰胺75 mg/kg腹腔注射,复制卵巢储备功能减退(DOR)大鼠模型。经灌胃治疗后,阴道涂片观察大鼠动情周期变化、酶联免疫法(ELISA)检测血清性激素、苏木素-伊红(HE)染色观察卵巢组织形态、荧光定量聚合酶链反应法(RT-qPCR)检测大鼠卵巢转化生长因子-β1(TGF-β1)、Smad2、Smad3、Smad7的基因表达。结果与空白组比较,膜型组大鼠的动情周期丧失规律性,血清雌二醇(E2)、抗缪勒管激素(AMH)水平显著降低(P<0.01),促卵泡激素(FSH)、促黄体生成素(LH)水平显著升高(P<0.01),卵巢指数、子宫指数明显降低(P<0.05),卵泡发育出现闭锁,卵巢中TGF-β1、Smad2、Smad3的表达量均减少(P<0.01),Smad7表达量增加(P<0.01);与模型组比较,中药低、高剂量组大鼠在给予药物治疗后动情周期逐渐恢复血清性激素、卵巢指数、子宫指数水平明显好转,卵巢中TGF-β1、Smad2、Smad3的表达量均增加(P<0.01),Smad7表达量减少(P<0.01)。结论补肾活血方中药能有效治疗DOR模型大鼠,可能是通过调节TGF-β1/Smads信号通路发挥作用。

Objective To observe the effects of Jin’s Bushen HuoxueDecoction on sex hormones and ovarian histomorphology in rats with ovarian reserve dysfunction,and the possible mechanism of regulating transforming growth factor-β1(TGF-β1)/Smads signal pathway.Methods Fifty female SD rats were randomly divided into 5 groups:blank group,model group,western medicine group,low dose group and high dose group,10 rats in each group.Except blank group,all the other groups were injected with cyclophosphamide 75 mg/kg intraperitoneally to replicate the rat model of ovarian reserve dysfunction(DOR).After intragastric administration,the changes of estrous cycle were observed by vaginal smear,serum sex hormones were detected by ELISA,the morphology of ovary was observed by hematoxylin-eosin(HE)staining,and the gene expressions of transforming growth factor-β1(TGF-β1),Smad2,Smad23,Smad27 were detected by fluorescence quantitative polymerase chain reaction(RT-qPCR).Results Compared with blank group,the estrous cycle of rats in model group lost regularity,the levels of serum estradiol(E2)and anti-Mullerian hormone(AMH)decreased significantly,the levels of follicle-stimulating hormone(FSH)and luteinizing hormone(LH)increased significantly,ovarian index and uterine index decreased significantly,and follicular development appeared atresia.The expression of TGF-β1,Smad2,Smad3 in ovary decreased,while the expression of Smad7 increased.Compared with model group,the estrous cycle of rats in low and high dose groups recovered gradually,the levels of serum sex hormone,ovarian index and uterine index were significantly improved,and the expression of TGF-β1,Smad2,Smad3 in ovary increased,while the expression of Smad7 decreased.Conclusion Bushen Huoxue Decoction can effectively treat DOR model rats,probably by regulating TGF-β1/Smads signal pathway.