Huang Tao, Gao Yang, Cao Yanwei, Wang Qinghai, Dong Zhen
Department of Kidney Transplantation, The Affiliated Hospital of Qingdao University, Qingdao, Shandong Province, PR China.
Department of Anesthesiology, The Affiliated Hospital of Qingdao University, Qingdao, Shandong Province, PR China.
Int Immunopharmacol. 2022 May;106:108614. doi: 10.1016/j.intimp.2022.108614. Epub 2022 Feb 12.
Acute kidney injury (AKI) is a widely pathophysiological state triggered by renal ischemia-reperfusion injury (IRI) during kidney transplant. Circular RNAs (circRNAs) have recently been shown to exert crucial roles in IRI. However, the underlying molecular mechanism is mainly undefined.
Differentially expressed circRNAs between IRI and sham group were identified by analyzing RNA-sequencing data in mice. Next, in vitro functional experiments were carried out to investigate the role of mmu_circ_0000943 in mouse kindey proximal tubule cell (TKPTS) apoptosis, inflammation response and oxidative stress using CCK-8, flow cytometry and ELISA assays, respectively. Moreover, bioinformatic prediction, western blot, luciferase reporter assay and RNA immunoprecipitation (RIP) were performed to examine the network among mmu_circ_0000943, miR-377-3p and early growth response 2 (Egr2).
Mmu_circ_0000943 was upregulated in renal IRI tissues and hypoxia/reoxygenation (H/R)-treated TKPTS cells. Knockdown of mmu_circ_0000943 inhibited cell apoptosis, inflammatory cytokine expression and oxidative stress upon H/R treatment. Mechanistically, co-transfection of siRNA targeting mmu_circ_0000943 and miR-377-3p inhibitor could counteract the anti-IRI effect. Furthermore, mmu_circ_0000943 regulated the expression of Egr2 by sponging miR-377-3p to alleviate H/R-induced TKPTS cell damage.
This study suggested that mmu_circ_0000943 participated in progression of renal IRI by sponging miR-377-3p with Egr2, providing a new insight into AKI treatment.
急性肾损伤（AKI）是一种广泛存在的病理生理状态，由肾移植过程中的肾缺血再灌注损伤（IRI）引发。环状RNA（circRNAs）最近被证明在IRI中发挥关键作用。然而，其潜在的分子机制主要尚不清楚。
通过分析小鼠的RNA测序数据，鉴定IRI组和假手术组之间差异表达的circRNAs。接下来，分别使用CCK-8、流式细胞术和ELISA检测，进行体外功能实验，以研究mmu_circ_0000943在小鼠肾近端小管细胞（TKPTS）凋亡、炎症反应和氧化应激中的作用。此外，进行生物信息学预测、蛋白质免疫印迹、荧光素酶报告基因检测和RNA免疫沉淀（RIP），以研究mmu_circ_0000943、miR-377-3p和早期生长反应2（Egr2）之间的网络关系。
mmu_circ_0000943在肾IRI组织和缺氧/复氧（H/R）处理的TKPTS细胞中上调。敲低mmu_circ_0000943可抑制H/R处理后的细胞凋亡、炎性细胞因子表达和氧化应激。机制上，共转染靶向mmu_circ_0000943的siRNA和miR-377-3p抑制剂可抵消抗IRI作用。此外，mmu_circ_0000943通过海绵吸附miR-377-3p来调节Egr2的表达，以减轻H/R诱导的TKPTS细胞损伤。
本研究表明，mmu_circ_0000943通过与Egr2结合海绵吸附miR-377-3p参与肾IRI的进展，为AKI治疗提供了新的见解。
