摘要
目的:观察预防性应用坤泰胶囊对环磷酰胺致卵巢储备功能低下大鼠血清性激素FSH、LH、E2、AMH水平的影响以及对卵巢GDF-9(生长分化因子9)、Egr-1(早期生长反应基因)基因及蛋白表达的影响,探讨坤泰胶囊对环磷酰胺致卵巢储备功能低下大鼠的治疗作用及作用机制。方法:将60只12周龄SD雌性大鼠随机分为空白组、环磷酰胺模型组(模型组)、坤泰胶囊组(分设高中低剂量)、结合雌激素组。空白组正常喂养;模型组用环磷酰胺腹腔注射法建立卵巢储备功能低下大鼠模型^([1]);各给药组于造模同时分别给予不同剂量的坤泰胶囊及结合雌激素片灌胃。实验期间观察大鼠行为学改变,行阴道涂片观察大鼠动情周期的变化;采用酶联免疫吸附法(ELISA)检测大鼠血清性激素AMH及FSH、LH、E2水平变化;光镜下观察卵巢形态与卵泡数;实时荧光定量(RT-PCR)检测大鼠卵巢GDF-9、Egr-1m RNA表达水平的变化;免疫组化法检测大鼠卵巢GDF-9、Egr-1蛋白表达水平的变化。结果:坤泰胶囊能够降低模型大鼠体内性激素FSH、LH的水平,升高E2及AMH水平,并且能够增加大鼠卵巢中GDF-9、EGR-1蛋白及m RNA表达,修复大鼠受损的卵巢组织结构,改善卵巢血供,减少卵泡闭锁,提高成熟卵泡存在数量,改善大鼠动情周期及行为学变化。结论:预防性应用坤泰胶囊,可有效提升卵巢敏感性,治疗效果与西药结合雌激素片无明显差异。
Objective:To observe the influences of different doses of Kuntai capsules on the level of serum sex hormones FSH,LH,E2,AMH and the ovarian GDF-9(growth differentiation factor-9),Egr-1(early growth response gene-1)gene and protein expression in rats with diminished ovarian reserve function due to cyclophosphamide,and to explore the therapeutic effect and mechanism of Kuntai capsules on rats with diminished ovarian reserve function due to cyclophosphamide Methods:60 female SD rats of 12-weeks old were randomly divided into blank group,cyclophosphamide model group(model group),Kuntai capsule group(high,middle and low dose),and conjugated estrogen group.The blank group was fed normally.The model group was modeled into rat model with diminished ovarian reserve function by intraperitoneal injection of cyclophosphamide.At the same time of modeling,each administration group was given the different doses of Kuntai capsules respectively and the conjugated estrogens tablets by gavage.During the experiment,the behavioral change of rats was observed and the change of rats'estrus cycle was observed by vaginal smear.The level changes of sex hormone AMH and FSH,LH,E2 in rats'serum were detected by enzyme-linked immunosorbent assay(ELISA).The ovarian morphology and follicle number were observed under light microscope.The changes of expression level of rats'ovarian GDF-9(growth differentiation factor-9),Egr-1(early growth response gene-1)and mRNA were detected by real-time fluorescence quantitative(RT-PCR).The changes of protein expression level of rats'ovarian GDF-9 and Egr-1 were measured by immunohistochemistry.Results:Kuntai capsules could lower the level of sex hormone FSH,LH in model rats,enhance the level of E2 and AMH,and increase GDF-9,Egr-1 protein in rats'ovarian and mRNA expression,repair the damaged ovarian tissue structure in rats,improve the blood supply for ovarian,reduce the follicular atresia,increase the existing number of mature follicles,improve the changes of estrous cycle and behavior of rats.Conclusion:In curative effect,Kuntai capsule has not obvious difference from conjugated estrogens tablet of western medicine.
作者
崔亚萌
高慧
徐文君
张飒
李欣捧
CUI Ya-meng;GAO Hui;XU Wen-jun;ZHANG Sha;LI Xin-peng(North China University of Science and Technology,Tangshan,063000,China)
出处
《山西中医》
2018年第3期47-51,共5页
Shanxi Journal of Traditional Chinese Medicine
基金
河北省中医药管理局中医药类科研计划课题(编号:2016068)
国家中医药管理局全国名老中医药专家高慧传承工作室建设项目(编号:2015048157)
河北省省级重大医学科研项目(编号:zyzd2013018)
河北省政府资助临床医学优秀人才培养项目(编号:2014571024)
关键词
卵巢储备功能低下
坤泰胶囊
结合雌激素
性激素
生长分化因子9
早期生长反应基因
免疫组化法
实时荧光定量
实验研究
diminished ovarian reserve function,Kuntai capsule,conjugated estrogens,sex hormone,growth differentiation factor-9,early growth response gene-1,immunohistochemistry,real-time fluorescence quantitative,experimental research
