Department of Endocrinology, Xuanwu Hospital, Capital Medical University, Beijing, China.
J Cell Physiol. 2019 Feb;234(2):1469-1476. doi: 10.1002/jcp.26959. Epub 2018 Jul 27.
Circular RNAs (circRNAs) are a novel type of noncoding RNAs that modulate the pathogenesis of multiple diseases. Nevertheless, the role of circRNAs in diabetic nephropathy (DN) pathogenesis is still ambiguous. In the current study, our team aims to investigate the expression profiles of circRNAs in DN and identify the function of circRNA on mesangial cells. CircRNAs microarray analysis revealed dysregulated circRNA in db/db DN mice, and circRNA_15698 was validated to be upregulated in both db/db mice and mouse mesangial cells (SV40-MES13) that were exposed to high glucose (25 mM) using real-time polymerase chain reaction. Loss-of-functional experiments showed that circRNA_15698 knockdown significantly inhibited the expression levels of collagen type I (Col. I), collagen type IV (Col. IV), and fibronectin. Moreover, the cellular localization of circRNA_15698 was mainly in the cytoplasm. Bioinformatics tools and luciferase reporter assay confirmed that circRNA_15698 acted as a 'sponge' of miR-185, and then positively regulated the transforming growth factor-β1 (TGF-β1) protein expression, suggesting a circRNA_15698/miR-185/TGF-β1 pathway. Further validation experiments validated that circRNA_15698/miR-185/TGF-β1 promoted extracellular matrix (ECM)-related protein synthesis. In summary, our study preliminarily investigates the role of circRNAs in mesangial cells and ECM accumulation, providing a novel insight for DN pathogenesis.
环状 RNA(circRNAs)是一种新型的非编码 RNA,可调节多种疾病的发病机制。然而,circRNAs 在糖尿病肾病(DN)发病机制中的作用仍不清楚。在本研究中,我们团队旨在研究 circRNAs 在 DN 中的表达谱,并鉴定 circRNA 对系膜细胞的功能。circRNA 微阵列分析显示 db/db DN 小鼠中存在失调的 circRNA,并用实时聚合酶链反应验证了在 db/db 小鼠和暴露于高葡萄糖(25mM)的小鼠系膜细胞(SV40-MES13)中circRNA_15698 上调。功能丧失实验表明,circRNA_15698 敲低显著抑制了 I 型胶原(Col. I)、IV 型胶原(Col. IV)和纤连蛋白的表达水平。此外,circRNA_15698 的细胞定位主要在细胞质中。生物信息学工具和荧光素酶报告基因检测证实,circRNA_15698 作为 miR-185 的“海绵”,并正向调节转化生长因子-β1(TGF-β1)蛋白表达,提示存在 circRNA_15698/miR-185/TGF-β1 通路。进一步的验证实验验证了 circRNA_15698/miR-185/TGF-β1 促进细胞外基质(ECM)相关蛋白的合成。总之,本研究初步研究了 circRNAs 在系膜细胞和 ECM 积累中的作用,为 DN 的发病机制提供了新的见解。