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微RNA-21和Peli1在自身免疫性早发性卵巢功能不全小鼠体内的表达及意义
杨雨涛,李欣然,谢嘉欣,王袁,马闻擎,付霞霏*
0
(南方医科大学珠江医院妇产科,广州 510280
*通信作者)
摘要:
目的 探讨miRNA-21和其靶基因Pellino-1(Peli1)在自身免疫性早发性卵巢功能不全(POI)模型小鼠体内的表达和意义。方法 将24只雌性BALB/c小鼠随机均分为POI组和对照组(每组12只),分别予皮下注射透明带3(ZP3)多肽和生理盐水。通过卵泡计数、观察发情周期和检测血清雌二醇、卵泡刺激素(FSH)、抗米勒管激素(AMH)水平评估小鼠卵巢功能。采用ELISA测定血清炎症因子γ-干扰素、IL-10水平,流式细胞术分析脾脏调节性T细胞(Treg)比例,生物信息学预测miRNA-21的靶基因,qPCR检测外周血单个核细胞(PBMC)、卵巢组织中miRNA-21表达和PBMC、脾脏Treg中Peli1的mRNA表达。结果 二次免疫6周后,POI组小鼠基本无发情期,而对照组小鼠发情周期均正常。POI组小鼠卵巢中各级卵泡的数量,血清雌二醇、AMH、IL-10水平,脾脏Treg比例,以及PBMC、卵巢组织中miRNA-21表达均低于对照组(P均 < 0.05),而血清FSH、γ-干扰素水平均高于对照组(P均 < 0.01)。生物信息学预测显示Peli1为miRNA-21的靶基因。POI组小鼠PBMC、脾脏Treg中Peli1的mRNA表达均低于对照组(P均 < 0.05)。结论 miRNA-21和其靶基因Peli1在自身免疫性POI模型小鼠中的表达均降低,提示miRNA-21和Peli1可能参与自身免疫性POI的发病。
关键词:  微RNA-21  Pellino-1  早发性卵巢功能不全  自身免疫性
DOI:10.16781/j.CN31-2187/R.20201155
投稿时间:2020-09-01
基金项目:广东省自然科学基金(2020A1515010205,2021A1515010701)
Expression and significance of microRNA-21 and Peli1 in mice with autoimmune premature ovarian insufficiency
YANG Yu-tao,LI Xin-ran,XIE Jia-xin,WANG Yuan,MA Wen-qing,FU Xia-fei*
(Department of Obstetrics and Gynaecology, Zhujiang Hospital of Southern Medical University, Guangzhou 510280, Guangdong, China
*Corresponding author)
Abstract:
Objective To investigate the expression and significance of microRNA-21 (miRNA-21) and its target gene Pellino-1 (Peli1) in mouse model of autoimmune premature ovarian insufficiency (POI).Methods Twenty-four female BALB/c mice were randomly divided into POI group and control group (12 in each group), with zona pellucida 3 (ZP3) polypeptide and the same dose of normal saline subcutaneously injected, respectively. Ovarian function was evaluated by counting follicles, observing estrus cycle and detecting the levels of serum estradiol, follicle-stimulating hormone (FSH) and anti-Müllerian hormone (AMH). The levels of interferon-γ and interleukin (IL)-10 were detected by enzyme-linked immunosorbent assay, the ratio of regulatory T cells (Tregs) of the spleen was detected by flow cytometry, the target gene of miRNA-21 was predicted by bioinformatics, and the expression of miRNA-21 in peripheral blood mononuclear cells (PBMCs) and ovarian tissues and the mRNA expression of Peli1 in PBMCs and Tregs of the spleen were detected by quantitative polymerase chain reaction.Results Six weeks after the second immunization, the mice in the POI group basically had no estrus, while the estrus cycle was normal in the control group. The number of follicles of each grade in the ovaries, the levels of serum estradiol, AMH and IL-10, ratio of Tregs in the spleen, and the expression levels of miRNA-21 in PBMCs and ovarian tissues were significantly lower in the POI group than those in the control group (all P < 0.05), while the levels of serum FSH and interferon-γ were significantly higher than those in the control group (both P < 0.01). Bioinformatics prediction revealed that Peli1 was a target gene of miRNA-21. The mRNA expression levels of Peli1 in PBMCs and splenic Tregs in the POI group were significantly lower than those in the control group (all P < 0.05).Conclusion The expression of miRNA-21 and its target gene Peli1 is significantly decreased in autoimmune POI mouse model, suggesting that miRNA-21 and Peli1 may be involved in the pathogenesis of autoimmune POI.
Key words:  microRNA-21  Pellino-1  premature ovarian insufficiency  autoimmune