WNT5A基因对卵巢颗粒细胞胰岛素反应性和胰岛素抵抗的调控作用

doi:10.6040/j.issn.1671-7554.0.2021.0304

摘要/Abstract

摘要： 目的探讨WNT5A基因过表达与颗粒细胞胰岛素抵抗发生的相关性。方法选取3周龄(n=15只)和6周龄(n=16只)雌性昆明小鼠,采用免疫组织化学和实时荧光定量聚合酶链反应(qRT-PCR)检测Wnt5a在小鼠卵巢颗粒细胞中的表达特征;通过Forskolin+pma模拟促黄体生成素( LH)峰处理人的卵巢颗粒细胞系24 h,检测LH峰作用后颗粒细胞的WNT5A表达对胰岛素反应性的影响;利用p-EGFP-N1-WNT5A转染KGN细胞系,检测过表达WNT5A与颗粒细胞胰岛素抵抗发生的相关性。结果与动情前期相比,Wnt5a在6周小鼠动情期(n=3,F=8.874,P=0.017)和动情后期(n=3,F=8.874,P=0.001)的卵巢中mRNA转录水平升高,在成熟卵泡颗粒细胞中蛋白表达水平升高。与未促排卵处理相比,促排卵处理可促进卵巢颗粒细胞Wnt5a mRNA转录水平升高(n=3,F=12.093,P=0.003),抑制KGN细胞的胰岛素反应性,而WNT5A则改善其胰岛素反应性。与转染质粒空载处理相比,过表达WNT5A导致胰岛素通路反应性降低,乳酸代谢降低(n=3,F=15.281,P<0.001)。结论卵巢排卵过程中WNT5A的表达水平升高,有利于增强颗粒细胞对胰岛素的反应性;而类似在多囊卵巢综合征(PCOS)患者中WNT5A表达异常升高时,可能会引起PCOS相关的胰岛素抵抗的发生。

关键词: 颗粒细胞, WNT5A基因, 过表达, 胰岛素抵抗

Abstract: Objective To explore the correlation between WNT5A gene overexpression and insulin resistance in granulosa cells. Methods Female Kunming mice aged 3 weeks(n=15)and 6 weeks(n=16)were selected. Expression of WNT5A in mice granulosa cells was detected with immunohistochemistry and real-time quantitative polymerase chain reaction(qRT-PCR). After human granulosa-like tumor cell line(KGN)were treated with Forskolin+pma to simulate luteinizing hormone(LH)peak for 24h, the effect of WNT5A on insulin reactivity was detected. After the KGN cells were transfected with p-EGFP-N1-WNT5A, the correlation between overexpression of WNT5A and insulin resistance was detected. Results Compared with preoestrus mice, oestrus mice (n=3, F=8.874, P=0.017)and postoestrus mice(n=3, F=8.874, P=0.001)had increased mRNA expression of WNT5A, and the protein expression of WNT5A increased in mature follicular granulosa cells. The expression of WNT5A in the granulosa cells of the ovulation-induced immature mice was also significantly increased(n=3, F=12.093, P=0.003). Ovulation induction inhibited insulin reactivity in KGN cells, which was improved by WNT5A. However, overexpression of WNT5A decreased insulin pathway responsiveness and lactate metabolism(n=3, F=15.281, P<0.001). Conclusion The expression of WNT5A benefits the insulin reactivity in granulosa cells during ovulation induction. However, when WNT5A is excessively elevated, such as in polycystic ovarian syndrome(PCOS), it may cause insulin resistance.

Key words: Granulosa cells, WNT5A gene, Overexpression, Insulin resistance

中图分类号:

R715.9

牛群,石婧婧,符江. WNT5A基因对卵巢颗粒细胞胰岛素反应性和胰岛素抵抗的调控作用[J]. 山东大学学报 (医学版), 2021, 59(6): 57-63.

NIU Qun, SHI Jingjing, FU Jiang. Effects of WNT5A gene on insulin reactivity and insulin resistance in granulosa cells[J]. Journal of Shandong University (Health Sciences), 2021, 59(6): 57-63.

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