Circ-ADAM9 Knockdown Reduces Insulin Resistance and Placental Injury in Diabetic Mice via MAPK Pathway Inactivation

IF 2.5 3区医学 Q3 IMMUNOLOGY

American Journal of Reproductive Immunology Pub Date : 2024-11-22 DOI:10.1111/aji.70017

Ai Zhao, Yawen Yang, Yijun Yang, Zhenjing Chi, Yanlan Sun

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引用次数: 0

Abstract

Background

Gestational diabetes mellitus (GDM) significantly risks maternal and neonatal health. Circular RNAs (circRNAs) regulate various diseases but their role in GDM is unclear. We investigated the involvement of circ-ADAM9 in GDM.

Methods

We analyzed circ-ADAM9 expression in GDM-related microarray data (GSE182737) and measured its levels in the blood of GDM patients. In a high-fat diet-induced GDM mouse model, we inhibited circ-ADAM9 expression and tracked blood glucose levels, serum insulin, lipid levels, placental apoptosis, and reactive oxygen species (ROS) levels. Pathological changes in pancreatic tissues and fetal outcomes were also examined. Molecular interactions were explored using bioinformatics tools and validated through luciferase assays, real-time quantitative polymerase chain reaction (RT-qPCR), and Western blotting in high glucose (HG)-induced human trophoblast cells (HTR-8/SVneo). We further investigated the involvement of circ-ADAM9/miR-375/FPR2 axis in HG-induced injury in HTR-8/SVneo cells by assessing cell viability, apoptosis, ROS production, and antioxidant levels.

Results

Both GDM patients and GDM-induced mice exhibited a substantial upregulation of circ-ADAM9. Knockdown of circ-ADAM9 lowered blood glucose, alleviated insulin resistance, improved lipid metabolism, decreased placental apoptosis and ROS levels, and reduced pancreatic lesions in GDM mice. Circ-ADAM9 downregulation also improved fetal viability, weight, and crown-rump length. In HG-induced HTR-8/SVneo cells, circ-ADAM9 overexpression and miR-375 downregulation were evident. Overexpression of miR-375 inhibited circ-ADAM9 activity, substantiating their binding interaction. In GDM mice, circ-ADAM9 deficiency restored miR-375 expression. TargetScanHuman predicted and luciferase assays confirmed the miR-375-FPR2 interaction and elevated FPR2 levels in GDM mice were reduced by circ-ADAM9 silencing. In HG-induced HTR-8/SVneo cells, circ-ADAM9 knockdown restored cell viability, suppressed apoptosis and ROS levels, and enhanced antioxidant enzyme levels. These effects were reversed by miR-375 inhibition or FPR2 overexpression, suggesting circ-ADAM9 upregulates FPR2 expression by sponging miR-375 and modulating the MAPK pathway.

Conclusion

This study is the first to demonstrate the expression and function of circ-ADAM9 in the progression of GDM. Circ-ADAM9 downregulation ameliorates insulin resistance and placental injury in GDM by modulating the miR-375/FPR2 axis and inactivating the MAPK pathway, which may offer a novel therapeutic target for the treatment of GDM.

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Circ-ADAM9 基因敲除通过 MAPK 通路失活减轻糖尿病小鼠的胰岛素抵抗和胎盘损伤

背景：妊娠糖尿病（GDM）严重危害孕产妇和新生儿的健康。环状 RNA（circRNA）可调控多种疾病，但在 GDM 中的作用尚不清楚。我们研究了 circ-ADAM9 在 GDM 中的作用：我们分析了 GDM 相关芯片数据（GSE182737）中 circ-ADAM9 的表达，并测量了 GDM 患者血液中的其水平。在高脂饮食诱导的 GDM 小鼠模型中，我们抑制了 circ-ADAM9 的表达，并跟踪了血糖水平、血清胰岛素、血脂水平、胎盘凋亡和活性氧（ROS）水平。我们还研究了胰腺组织的病理变化和胎儿的结局。我们使用生物信息学工具探索了分子相互作用，并通过荧光素酶测定、实时定量聚合酶链反应（RT-qPCR）和高葡萄糖（HG）诱导的人滋养层细胞（HTR-8/SVneo）的 Western 印迹进行了验证。我们通过评估细胞活力、凋亡、ROS生成和抗氧化剂水平，进一步研究了circ-ADAM9/miR-375/FPR2轴在HG诱导的HTR-8/SVneo细胞损伤中的参与情况：结果：GDM 患者和 GDM 诱导的小鼠都表现出 circ-ADAM9 的大量上调。敲除 circ-ADAM9 能降低 GDM 小鼠的血糖、缓解胰岛素抵抗、改善脂质代谢、减少胎盘凋亡和 ROS 水平并减少胰腺病变。下调 Circ-ADAM9 还能提高胎儿存活率、体重和头臀长。在 HG 诱导的 HTR-8/SVneo 细胞中，circ-ADAM9 的过表达和 miR-375 的下调是显而易见的。miR-375的过表达抑制了circ-ADAM9的活性，证实了它们之间的结合相互作用。在 GDM 小鼠中，circ-ADAM9 的缺乏可恢复 miR-375 的表达。TargetScanHuman 预测和荧光素酶测定证实了 miR-375-FPR2 的相互作用，并且通过沉默 circ-ADAM9 降低了 GDM 小鼠中升高的 FPR2 水平。在 HG 诱导的 HTR-8/SVneo 细胞中，circ-ADAM9 基因敲除可恢复细胞活力，抑制细胞凋亡和 ROS 水平，提高抗氧化酶水平。这些效应被miR-375抑制或FPR2过表达所逆转，表明circ-ADAM9通过疏导miR-375和调节MAPK通路上调FPR2的表达：本研究首次证明了 circ-ADAM9 在 GDM 进展过程中的表达和功能。通过调节miR-375/FPR2轴和失活MAPK通路，下调circ-ADAM9可改善GDM中的胰岛素抵抗和胎盘损伤，这可能为治疗GDM提供一个新的治疗靶点。

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来源期刊

American Journal of Reproductive Immunology 医学-免疫学

CiteScore

6.20

自引率

5.60%

发文量

314

审稿时长

2 months

期刊介绍： The American Journal of Reproductive Immunology is an international journal devoted to the presentation of current information in all areas relating to Reproductive Immunology. The journal is directed toward both the basic scientist and the clinician, covering the whole process of reproduction as affected by immunological processes. The journal covers a variety of subspecialty topics, including fertility immunology, pregnancy immunology, immunogenetics, mucosal immunology, immunocontraception, endometriosis, abortion, tumor immunology of the reproductive tract, autoantibodies, infectious disease of the reproductive tract, and technical news.