基于Kisspeptin/GPR54系统探讨新加二甲地黄汤对PCOS模型大鼠卵泡发育的影响OA
Exploring the effect of Xinjia Erjia Dihuang Tang on follicular development in PCOS model rats based on the Kisspeptin/GPR54 system
目的 探讨经新加二甲地黄汤干预后的多囊卵巢综合征(polycystic ovary syndrome,PCOS)模型大鼠的卵泡发育情况及其可能存在的效应机制.方法 筛选 28 只拥有规律动情周期的雌性SD大鼠,随机分为正常对照组、模型组、中药组及西药组,每组 7 只.除正常对照组外,其余三组均连续予来曲唑-羧甲基纤维素混悬液 0.1mg/(kg·d)灌胃,以构建PCOS大鼠模型.自第 22 天起,中药组以新加二甲地黄汤 5.268g/(kg·d)灌胃,西药组以炔雌醇环丙孕酮片0.286mg/(kg·d)灌胃,正常对照组及模型组均以 10ml/(kg·d)蒸馏水灌胃.3 周后比较各组大鼠卵巢系数,苏木精-伊红染色观察各组大鼠卵巢组织形态学改变,对各组大鼠血清激素均采用酶联免疫吸附试验进行检测,蛋白质印迹法检测卵巢亲吻素(kisspeptin,Kp)、G蛋白偶联受体 54(G-protein-coupled receptor 54,GPR54)蛋白表达水平.结果 与正常对照组比较,模型组大鼠卵巢内呈现为囊状扩张和闭锁的卵泡增多,其颗粒细胞层数变少,卵巢系数增大;血清睾酮(testosterone,T)、黄体生成素(luteinizing hormone,LH)、Kp水平升高;血清卵泡刺激素(follicle stimulating hormone,FSH)、雌二醇(estradiol,E2)水平及大鼠卵巢组织中Kp、GPR54 蛋白表达均显著降低(P<0.05).予中药干预后,与模型组比较,中药组大鼠卵巢囊样扩张卵泡数量变少,颗粒细胞的层数增多,存在近成熟的卵泡,并见少量黄体存在;大鼠血清LH、T、Kp水平下降,血清FSH、E2水平及卵巢Kp、GPR54 蛋白表达显著升高(P<0.05).结论 PCOS模型大鼠的卵泡发育情况经新加二甲地黄汤干预后得以改善,该治疗机制可能为通过调控Kisspeptin/GPR54 系统影响FSH和LH的释放,以此影响激素含量,调整卵巢功能,使卵泡发育和排卵能力得以改善.
Objective To investigate the development of follicles in polycystic ovary syndrome(PCOS)model rats treated with Xinjia Erjia Dihuang Tang and its possible mechanism.Methods Twenty-eight female SD rats with regular estrous cycle were selected and randomly divided into normal control group,model group,Chinese medicine group and Western medicine group,with 7 rats in each group.In addition to the normal control group,the other three groups were continuously given 0.1mg/(kg·d)of letrozole-carboxymethyl cellulose suspension to construct the PCOS rat model.Since the 22nd day,the traditional Chinese medicine group was given 5.268g/(kg·d)of Xinjia Erjia Dihuang Tang,the western medicine group was given 0.286mg/(kg·d)of ethinylestradiol cyproterone tablet,and the normal control group and model group were given 10ml/(kg·d)of distilled water.After 3 weeks,the ovarian coefficients of the rats in all groups were compared,and the morphological changes of the ovarian tissues of the rats in all groups were observed by hematoxylin-eosin staining.The serum hormones of the rats in all groups were detected by enzyme-linked immunosorbent assay,and the ovarian kisspeptin(Kp)and G-protein-coupled receptors(GPR54)protein expression levels were detected by Western blot.Results Compared with the normal control group,the follicles in the ovary of the model group showed more cystic dilatation and atresia,and the number of granular cell layers decreased,the ovarian coefficient of the model group was increased;Serum levels of testosterone(T),luteinizing hormone(LH)and Kp were increased in model group;Serum levels of follicle stimulating hormone(FSH)and estradiol(E2)and protein expressions of Kp and GPR54 in ovarian tissue of rats in the model group were significantly decreased(P<0.05).After treatment with Chinese medicine,compared with the model group,the number of follicles in the Chinese medicine group decreased,the layer number of granulosa cells increased,there were nearly mature follicles,and a small amount of corpus luteum existed,serum LH,T and Kp levels were decreased,serum FSH and E2 levels and ovarian Kp and GPR54 protein expressions were significantly increased(P<0.05).Conclusion The development of follicles in PCOS model rats was improved by the intervention of Xinjia Erjia Dihuang Tang.The therapeutic mechanism may be to affect the release of FSH and LH by regulating Kisspeptin/GPR54 system,so as to affect hormone content,adjust ovarian function,and improve follicle development and ovulation ability.
石明晴;王津;徐小雨;蓝关翠
丽水市中医院妇产科,浙江丽水 323000浙江中医药大学第三临床医学院,浙江杭州 310053
临床医学
新加二甲地黄汤多囊卵巢综合征G蛋白偶联受体54Kisspeptin性激素
Xinjia Erjia Dihuang TangPolycystic ovary syndromeG-protein-coupled receptor 54KisspeptinSex hormones
《中国现代医生》 2024 (018)
90-95 / 6
浙江省中医药科学研究基金项目(2021ZB338);浙江省卫生健康科技计划项目(2021RC147)
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