Yao Tao, Zha Dongqing, Gao Ping, Wu Xiaoyan
Department of Nephrology, Zhongnan Hospital of Wuhan University, Wuhan, China.
J Cell Mol Med. 2021 Jan 23;25(13):5940-8. doi: 10.1111/jcmm.16286.
Recent studies indicate that circular RNAs are involved in dysregulation of kidney injury. Nevertheless, the underlying mechanisms remain largely unclear. Therefore, this study sought to investigate the role of circ-USP1 in the pathogenesis of early renal allografts. Thirty-two male C57BL/6J mice aged between 6 and 8 weeks were divided into the sham and allograft groups. Thereafter, the association between miR-194-5p, circ-USP1 and DNMT3A was confirmed using a combination of bioinformatics and the luciferase reporter gene assay. Additionally, the expression of circ-USP1, miR-194-5p and DNMT3A mRNA was detected through qPCR. Afterwards, the Western blot assay was performed to examine the expression of DNMT3A protein. Finally, the TUNEL assay was conducted to determine the rate of apoptosis in DNMT3A cells. The expression of circ-USP1 increased, while that of miR-194-5p decreased in renal allografts. Additionally, silencing circ-USP1 reduced kidney injuries caused by renal allografts in mice. Moreover, miR-194-5p was a target for circ-USP1, and DNMT3A was a target of miR-194-5p. Finally, it was shown that silencing circ-USP1 reduced DNMT3A expression in the kidney of mice that received renal allografts. Circ-USP1 functions as a competing endogenous RNA for miR-194-5p. This occurs in order to regulate DNMT3A expression in kidney injury induced by hypoxia in acute renal allografts.
近期研究表明,环状RNA参与了肾损伤的失调。然而,其潜在机制仍 largely不清楚。因此,本研究旨在探讨环状-USP1在早期肾移植发病机制中的作用。将32只6至8周龄的雄性C57BL/6J小鼠分为假手术组和移植组。此后,结合生物信息学和荧光素酶报告基因检测,证实了miR-194-5p、环状-USP1和DNMT3A之间的关联。此外,通过qPCR检测环状-USP1、miR-194-5p和DNMT3A mRNA的表达。之后,进行蛋白质印迹分析以检测DNMT3A蛋白的表达。最后,进行TUNEL检测以确定DNMT3A细胞中的凋亡率。肾移植中环状-USP1的表达增加,而miR-194-5p的表达降低。此外,沉默环状-USP1可减轻小鼠肾移植引起的肾损伤。此外,miR-194-5p是环状-USP1的靶标,而DNMT3A是miR-194-5p的靶标。最后,结果表明,沉默环状-USP1可降低接受肾移植小鼠肾脏中DNMT3A的表达。环状-USP1作为miR-194-5p的竞争性内源RNA发挥作用。这发生在急性肾移植中缺氧诱导的肾损伤中,以调节DNMT3A的表达。