Circular RNA circRNA_15698 aggravates the extracellular matrix of diabetic nephropathy mesangial cells via miR-185/TGF-β1
Circular RNA circRNA_15698 aggravates the extracellular matrix of diabetic nephropathy mesangial cells via miR-185/TGF-β1
复制标题
DOI：
10.1002/jcp.26959
复制
发表时间：
2019-02-01
影响因子：
5.6
通讯作者：
Wang, Li
中科院分区：
文献类型：
--
作者：
Hu, Wei;Han, Qing;Wang, Li
环状RNA（circRNA）是一种新型非编码RNA，可调节多种疾病的发病机制。然而，circRNA在糖尿病肾病（DN）发病机制中的作用仍然不明确。在本研究中，我们的团队旨在研究 DN 中 circRNA 的表达谱并确定 circRNA 在系膜细胞上的功能。 CircRNA 微阵列分析显示 db/db DN 小鼠中的 circRNA 失调，并且使用实时聚合酶链式反应验证了 circRNA_15698 在暴露于高葡萄糖 (25 mM) 的 db/db 小鼠和小鼠系膜细胞 (SV40-MES13) 中表达上调。功能丧失实验表明，circRNA_15698 敲低显着抑制 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充当miR-185的“海绵”，然后正向调节转化生长因子-β1（TGF-β1）蛋白的表达，表明circRNA_15698/miR-185/TGF-β1途径。进一步的验证实验验证了circRNA_15698/miR-185/TGF-beta 1促进细胞外基质（ECM）相关的蛋白质合成。总之，我们的研究初步探讨了circRNA在系膜细胞和ECM积累中的作用，为DN发病机制提供了新的见解。
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-beta 1 (TGF-beta 1) protein expression, suggesting a circRNA_15698/miR-185/TGF-beta 1 pathway. Further validation experiments validated that circRNA_15698/miR-185/TGF-beta 1 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.