NPPA/NPR1信号促进小鼠卵泡扩张的作用机制  

Mechanism of NPPA/NPR1 signaling promoting ovarian follicle expansion in mice

作　　者：刘嘉禹 陈晓[1] 郝肖琼 LIU Jiayu;CHEN Xiao;HAO Xiaoqiong(Department of Physiology,Baotou Medical College,Baotou 014040,China;School of Medicine,South China Universi-ty of Technology,Guangzhou 510006,China)

机构地区：[1]包头医学院生理学教研室,内蒙古包头014040 [2]华南理工大学医学院,广东广州510006

出　　处：《中国病理生理杂志》2025年第4期714-722,共9页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.32360181);内蒙古自治区自然科学基金资助项目(No.2022LHMS03012);内蒙古自治区卫生健康科技计划项目(No.202201376)。

摘　　要：目的:探究利尿钠肽A/利尿钠肽受体1(NPPA/NPR1)信号在促黄体素(LH)诱导的小鼠卵泡扩张中的作用及机制。方法:3周龄雌性小鼠随机分为注射孕马血清促性腺激素(e CG)48 h组以及注射e CG后接着注射人绒毛膜促性腺激素(h CG)4和8 h组,每组6只,利用过碘酸希夫(PAS)染色观察卵泡直径变化,qPCR检测小鼠颗粒细胞中Npr1 m RNA表达情况。野生型(WT)小鼠及Npr1基因敲除(Npr1^(-/-))小鼠分别注射eCG 48 h或注射eCG48 h/hCG 4 h,每组6只,利用PAS染色观察卵泡直径变化。利用超数排卵和繁殖实验统计WT小鼠的排卵数及产仔数,排卵实验每组6只,繁殖实验每组7只。获取分离好的小鼠卵泡,随机分为对照组、NPPA组、LH组、LH+NPPA组、LH+NPPA+KT5823[蛋白激酶G(PKG)抑制剂]组及LH+NPPA+PPQ-102[囊性纤维化跨膜转导调节因子(CFTR)抑制剂]组,每组20枚卵泡,另取WT小鼠与Npr1^(-/-)小鼠分别注射eCG 48 h或注射eCG 48 h/h CG 4 h,每组3只,利用体外卵泡培养以及qPCR探究PKG和CFTR信号对NPPA/NPR1诱导的卵泡扩张的影响;利用免疫荧光染色检测WT及Npr1^(-/-)小鼠颗粒细胞增殖能力,每组5只。结果:注射h CG能够促进小鼠卵泡扩张,同时颗粒细胞中Npr1的m RNA表达显著上调(P<0.01);与WT小鼠相比,Npr1^(-/-)小鼠卵泡扩张显著受限,排卵数与产仔数均显著下降(P<0.01);PKG抑制剂KT5823和CFTR抑制剂PPQ-102能够显著抑制NPPA/NPR1诱导的卵泡扩张(P<0.01),且Npr1^(-/-)小鼠颗粒细胞中Pkg mRNA及Cftr mRNA表达与WT小鼠相比均显著下降(P<0.01)。另外,Npr1^(-/-)小鼠颗粒细胞增殖能力显著低于WT小鼠(P<0.01)。结论:LH信号通过NPPA/NPR1信号激活下游PKG、CFTR及颗粒细胞增殖途径,促进小鼠卵泡扩张。AIM:To explore the role and mechanism of natriuretic peptide A(NPPA)/natriuretic peptide receptor 1(NPR1)signaling in luteinizing hormone(LH)-induced ovarian follicle expansion in mice.METHODS:Threeweek-old female mice were injected with pregnant mare serum gonadotropin/equine chorionic gonadotrophin(eCG)48 h before use to stimulate follicle development,or injected with eCG followed by human chorionic gonadotropin(hCG)treatment for 4 or 8 h,with 6 mice in each group.Periodic acid-Schiff(PAS)staining in ovarian sections was used to observe the follicular diameter,and qPCR was used to detect the expression of Npr1 mRNA levels in granulosa cells.Wild-type(WT)and Npr1 knockout(Npr1^(-/-))mice were injected with eCG,or injected with eCG followed by hCG treatment for 4 h.PAS staining was used to observe the follicular diameter,with 6 mice in each group.Superovulation and fertility assays were performed to count the number of ovulated oocytes and pups,with 6 mice in each group subjected to superovulation and 7 mice in each group subjected to fertility assays.The isolated follicles were randomly divided into control,NPPA,LH,LH+NPPA,LH+NPPA+KT5823[protein kinase G(PKG)inhibitor],and LH+NPPA+PPQ-102[cystic fibrosis transmembrane conductance regulator(CFTR)inhibitor]groups,with 20 follicles in each group.In addition,WT and Npr1^(-/-)mice were injected with eCG 48 h or eCG 48 h/hCG 4 h,with 3 mice in each group.In vitro follicle culture and qPCR were used to explore the effects of PKG and CFTR signalling on NPPA/NPR1-induced follicular expansion.The proliferation of granulosa cells in WT and Npr1^(-/-)mice was detected via immunofluorescence staining,with 5 mice in each group.RESULTS:The results revealed that hCG treatment significantly increased the follicle expansion and Npr1 mRNA expression in granulosa cells(P<0.01).The capacity for ovarian follicle expansion,the number of ovulations,and the number of pups per litter were significantly lower in Npr1^(-/-)mice compared with WT mice(P<0.01).In in vitro follicular culture,t

关 键 词：卵泡扩张 利尿钠肽受体1 利尿钠肽A 小鼠 

分 类 号：R339.2[医药卫生—人体生理学] R363.2[医药卫生—基础医学] R711