摘要
目的探讨卵巢表达的kisspeptin/kiss1r系统在卵巢早衰小鼠模型中的表达变化及发挥的作用。方法选用6周龄雌性balb/c小鼠,随机分为正常组、对照组和POF组,其中POF组腹腔注射环磷酰胺(70mg/kg,0.01%DMSO),对照组腹腔注射等量的0.01%DMSO。1周后,酶联免疫吸附法检测3组小鼠血清中E2、FSH水平,HE染色观察卵巢的形态学改变,免疫组化检测小鼠卵巢中kisspeptin/kiss1r的表达变化,masson法检测小鼠卵巢中纤维情况。结果 ELISA结果显示与对照组相比,POF模型组小鼠血清中E2水平明显减少,而FSH水平显著升高。POF组小鼠卵巢中生长卵泡数量明显减少,卵泡中颗粒细胞的层数明显减少,闭锁卵泡数量明显增加;除卵巢间质外,卵泡内有大量的胶原纤维存在。免疫组化结果显示与对照组相比,POF小鼠卵巢次级卵泡中kisspeptin/kiss1r系统的阳性细胞数明显减少。结论POF小鼠卵巢表达的kisspeptin可能通过降低卵泡对FSH的敏感性,大量的初级卵泡被募集,最终使卵母细胞耗竭。
Objective To explore the expression of kisspeptin/kiss1 rsystem in the ovary and the role played during the premature ovarian failure model.Methods Female balb/c mice aged 6 weeks were divided randomly into the normal group,the control group and the POF group.POF mice were treated with Cyclophosphamide（70 mg/kg,suspended in 0.01% DMSO）and the control group received 0.01% DMSO by intraperitoneal injection.One week after the treatment,the serum levels of folliclestimulating hormone（FSH）and estradiol（E2）in three groups were assessed by enzyme-linked immunosorbent assay（ELISA）.Ovarian histopathological changes were evaluated by hematoxylin-eosin staining（HE staining）.Protein levels of kisspeptin/kiss1 rsystem in the ovary were evaluated by immunohistochemistry.The collagenous fiber was detected by masson method.Results ELISA result showed that compared with those in the control group,levels of E2 decreased and levels of FSH increased markedly in serum of POF mice.The number of growing follicles and the layer of granulosa cells decreased but atresia follicle increased in the ovaries of POF mice.A large amount of collagen fibers had appeared in the follicle in addition to ovarian stroma.Positive cells of kisspeptin/kiss1 rsystem in the secondary follicles decreased significantly in POF mice.ConclusionKisspeptin,expressed in POF mice ovaries,may make follicle reduce the sensitivity to FSH,so that a large number of primary follicles are raised,and eventually the oocyte is exhausted.
作者
师红美
刘冉冉
张洪芹
SHI Hongmei;LIU Ranran;ZHANG Hongqin(School of Basic Medical Sciences, Binzhou Medical University, Yantai 264003, P. R. China;Department of Radiology, Qilu Hospital of Shandong University（Qingdao};Department of Reproductive Medicine, Yantai Affiliated Hospital of Binzhou Medical University;Department of Histology and Emnryology, School of Basic Medical Sciences, Binzhou Medical University)
出处
《滨州医学院学报》
2017年第6期401-404,470,共5页
Journal of Binzhou Medical University
基金
滨州医学院科技计划项目(BY2016KJ36)
