Kidney fibrosis is a major mechanism underlying chronic kidney disease (CKD). N⁶-methyladenosine (m⁶A) RNA methylation is associated with organ fibrosis. We investigated m⁶A profile alterations and the inhibitory effect of RNA methylation in kidney fibrosis in vitro (TGF-β-treated HK-2 cells) and in vivo (unilateral ureteral obstruction [UUO] mouse model). METTL3-mediated signaling was inhibited using siRNA in vitro or the METTL3-specific inhibitor STM2457 in vivo and in vitro. In HK-2 cells, METTL3 protein levels increased in a dose- and time-dependent manner along with an increase in the cellular m⁶A levels. In the UUO model, METTL3 expression and m⁶A levels were significantly increased. Transcriptomic and m⁶A profiling demonstrated that epithelial-to-mesenchymal transition- and inflammation-related pathways were significantly associated with RNA m⁶A methylation. Genetic and pharmacologic inhibition of METTL3 in HK-2 cells decreased TGF-β-induced fibrotic marker expression. STM2457-induced inhibition of METTL3 attenuated the degree of kidney fibrosis in vivo. Furthermore, METTL3 protein expression was significantly increased in the tissues of CKD patients with diabetic or IgA nephropathy. Therefore, targeting alterations in RNA methylation could be a potential therapeutic strategy for treating kidney fibrosis.

肾脏纤维化是慢性肾脏病（CKD）的主要机制。 N ⁶ -甲基腺苷 (m ⁶ A) RNA 甲基化与器官纤维化相关。我们在体外（TGF-β处理的 HK-2 细胞）和体内（单侧输尿管梗阻 [UUO] 小鼠模型）研究了 m ⁶ A 谱的变化和 RNA 甲基化对肾纤维化的抑制作用。在体外使用 siRNA 或在体内和体外使用 METTL3 特异性抑制剂 STM2457 抑制 METTL3 介导的信号传导。在 HK-2 细胞中，METTL3 蛋白水平随着细胞 m ⁶ A 水平的增加而以剂量和时间依赖性方式增加。在UUO模型中，METTL3表达和m ⁶ A水平显着增加。转录组和 m ⁶ A 分析表明上皮间质转化和炎症相关途径与 RNA m ⁶ A 甲基化显着相关。 HK-2 细胞中 METTL3 的遗传和药理抑制可降低 TGF-β 诱导的纤维化标志物表达。 STM2457 诱导的 METTL3 抑制减轻了体内肾纤维化的程度。此外，患有糖尿病或IgA肾病的CKD患者组织中METTL3蛋白表达显着增加。因此，针对 RNA 甲基化的改变可能是治疗肾纤维化的潜在治疗策略。