Abstract：
Podocyte dysfunction has been identified as a crucial pathological characteristic of diabetic nephropathy (DN). However, the regulatory effects of long non-coding RNAs (lncRNAs) in this process have not been fully elucidated. Here, we performed an unbiased RNA-sequencing (RNA-seq) analysis of renal tissues and identified a significantly upregulated long non-coding RNA, ENST00000585189.1 (lncRNA 585189), in patients with DN. Furthermore, lncRNA 585189 was positively correlated with renal insufficiency and was upregulated in both DN patients and high-glucose-induced human podocytes. Gain- and loss-of-function experiments revealed that silencing lncRNA 585189 decreased the production of ROS, rescued aberrant mitochondrial morphology and membrane potential, and alleviated podocyte damage caused by high glucose. Mechanistically, bioinformatics analysis predicted an interaction between lncRNA 585189 and hnRNP A1, which was subsequently confirmed by RIP, pull-down, and EMSA assays. Further investigation revealed that lncRNA 585189 destabilizes the hnRNP A1 protein, leading to the downregulation of its expression. Conversely, hnRNP A1 promoted the expression of lncRNA 585189. Moreover, both RIP and pull-down assays demonstrated a direct interaction between hnRNP A1 and SIRT1, which enhanced SIRT1 mRNA stability. Our findings suggest that lncRNA 585189 suppresses SIRT1 through hnRNP A1, thereby hindering the recovery from mitochondrial abnormalities and podocyte damage. In summary, targeting lncRNA 585189 is a promising strategy for reversing mitochondrial dysfunction and treating DN.
摘要：
足细胞功能障碍已被确定为糖尿病肾病（DN）的重要病理特征。然而,长链非编码RNA(lncRNA)在这一过程中的调节作用尚未完全阐明.这里,我们对肾组织进行了无偏RNA测序（RNA-seq）分析，并确定了显着上调的长链非编码RNA，ENST00000585189.1(lncRNA585189),DN患者。此外，lncRNA585189与肾功能不全呈正相关，并且在DN患者和高糖诱导的人足细胞中均上调。功能的增益和丧失实验表明，沉默lncRNA585189降低了ROS的产生，拯救异常的线粒体形态和膜电位，减轻高糖引起的足细胞损伤。机械上,生物信息学分析预测了lncRNA585189和hnRNPA1之间的相互作用，随后被RIP证实，下拉,和EMSA检测。进一步的研究表明，lncRNA585189使hnRNPA1蛋白不稳定，导致其表达下调。相反，hnRNPA1增进了lncRNA585189的表达。此外，RIP和下拉测定均表明hnRNPA1和SIRT1之间存在直接相互作用,这增强了SIRT1mRNA的稳定性.我们的发现表明，lncRNA585189通过hnRNPA1抑制SIRT1，从而阻碍线粒体异常和足细胞损伤的恢复。总之,靶向lncRNA585189是逆转线粒体功能障碍和治疗DN的有前途的策略。
