滋阴温阳序贯方对卵巢功能减退动物模型的调节作用

Regulatory effect of the prescription for nourishing yin and warming yang in a sequential way on the animal model of ovarian

目的探寻滋阴温阳序贯方对卵巢功能减退动物模型的调节作用。方法将有规律动情周期SPF级SD健康雌性大鼠按照随机数法分为正常对照组、模型组、滋阴温阳序贯方药组(中药组)和西药组,共4组,每组8只。除模型组外均予雷公藤多甙片75 mg/kg混悬液2 mL,每日1次,灌胃,连续14 d,建立卵巢功能减退动物模型。西药组给予戊酸雌二醇片按0.09 mg/kg灌胃给药,中药组给予滋阴温阳序贯方药按10 mL/kg灌胃给药,每天灌胃给药1次,连续2周。分别于给药第1天、第7天和第14天检测卵巢系数变化。采用ELISA法检测大鼠血清促卵泡成熟激素(FSH)、促黄体生成素(LH)、雌二醇(E_(2))和抗缪勒管激素(AMH)的水平;HE检测卵巢组织病理学变化;通过免疫组织化学方法和免疫印迹(Western blot)手段检测IGF-1和SCF的蛋白表达情况。结果给药第14天,大鼠卵巢病理变化,与对照组比较,模型组小鼠各级卵泡数量减少,颗粒细胞完整度破坏;与模型组比较,西药组和中药组小鼠卵巢内可见新生卵泡,颗粒细胞层数增加。卵巢系数比较,西药组和中药组较对照组低,较模型组显著升高,差异有统计学意义(P<0.05)。E_(2)与AMH含量比较,西药组和中药组较模型组显著升高(P<0.05)。FSH和LH浓度含量比较,西药组和中药组较模型组显著降低(P<0.05)。西药组和中药组IGF-1表达量显著高于模型组(P<0.05);SCF表达量显著低于模型组(P<0.05)。结论滋阴温阳序贯方可以通过增加卵巢系数、卵泡数量和质量、改善激素分泌和细胞因子表达,进而改善卵巢功能。

Objective To explore the regulatory effect of the prescription for nourishing yin and warming yang in a sequential way on the animal model of ovarian hypofunction.Methods The SPF SD healthy female rats with a regular estrous cycle were randomly divided into a normal control group,a model group,a Chinese medicine group and a western medicine group,with 8 rats in each group.Except the model group,all rats were given the 2 mL suspension of tripterygium wilfordii polyglycoside tablets (75 mg/kg) by gavage once a day,for 14 consecutive days,to establish the animal model of ovarian hypofunction.The western medicine group was given estradiol valerate tablets (0.09 mg/kg) by gavage,while the Chinese medicine group was given the decoction (10 mL/kg) by gavage,once a day,for two consecutive weeks.The changes of ovarian coefficient were detected on the 1st,7th and 14th day after administration.The serum levels of follicle stimulating hormone (FSH),luteinizing hormone (LH),estradiol (E) and anti-Mullerian hormone (AMH) were measured by ELISA.The histopathological changes of ovary were detected by HE.The protein expression of IGF-1 and SCF was detected by the immunohistochemistry and Western blot.Results The ovarian coefficient of the western medicine group and the Chinese medicine group was lower than that of the control group,yet significantly higher than that of the model group (P<0.05).After the intervention of western medicine and Chinese medicine,the contents of Eand AMH were significantly higher than those in the model group,while the concentrations of FSH and LH were significantly lower than those in the model group (P<0.05).Compared with the control group,The 14th day of administration.The expression of IGF-1 was significantly higher than that of the model group after the intervention of western medicine and Chinese medicine.After the intervention of western medicine and Chinese medicine,the expression of SCF was significantly lower than that of the model group (P<0.05).Conclusion The prescription can improve the ovarian function by increasing the ovarian coefficient,follicle quantity and quality,improving the hormone secretion and cytokine expression.