Qu Gaoting, He Tiantian, Dai Aisuo, Zhao Yajie, Guan Dian, Li Shanwen, Shi Huimin, Gan Weihua, Zhang Aiqing
Department of Pediatric Nephrology, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210003, P.R. China.
Department of Pediatrics, Taizhou People's Hospital, Taizhou, Jiangsu 225300, P.R. China.
Exp Ther Med. 2021 Dec;22(6):1469. doi: 10.3892/etm.2021.10904. Epub 2021 Oct 21.
Podocyte apoptosis is a key risk factor for the progression of kidney diseases. MicroRNA (miR)-199b-5p has been shown to be involved in cell apoptosis. However, the molecular mechanisms of miR-199b-5p in podocyte apoptosis remain uncertain. Thus, the present study aimed to investigate whether miR-199b-5p participates in the regulation of podocyte apoptosis and to elucidate the involved mechanisms of this process. A podocyte apoptosis model was constructed using adriamycin (ADR) . miR-199b-5p mimic and inhibitor were transfected in podocytes to change the expression level of miR-199b-5p. RNA expression was examined by reverse transcription-quantitative PCR. Western blotting was used to measure protein expression. Apoptosis was monitored via flow cytometry and detection of apoptosis-associated proteins. The results from the present study demonstrated that miR-199b-5p was upregulated and that regulator of G-protein signaling 10 (RGS10) was downregulated in ADR-stimulated podocytes. Overexpression of miR-199b-5p could inhibit RGS10 expression and stimulate podocyte apoptosis, whereas miR-199b-5p knockdown restored the levels of RGS10 and ameliorated podocyte apoptosis in ADR-induced podocytes. Furthermore, the effects of miR-199b-5p overexpression could be significantly reversed by RGS10 overexpression. In addition, podocyte transfection of miR-199b-5p activated the AKT/mechanistic target of rapamycin (mTOR) signaling, which was blocked following RGS10 overexpression. Taken together, the present study demonstrated that miR-199b-5p upregulation could promote podocyte apoptosis by inhibiting the expression of RGS10 through the activation of AKT/mTOR signaling.
足细胞凋亡是肾脏疾病进展的关键危险因素。微小RNA（miR）-199b-5p已被证明参与细胞凋亡。然而，miR-199b-5p在足细胞凋亡中的分子机制仍不明确。因此，本研究旨在探讨miR-199b-5p是否参与足细胞凋亡的调控，并阐明这一过程的相关机制。使用阿霉素（ADR）构建足细胞凋亡模型。将miR-199b-5p模拟物和抑制剂转染到足细胞中以改变miR-199b-5p的表达水平。通过逆转录定量PCR检测RNA表达。采用蛋白质印迹法检测蛋白质表达。通过流式细胞术和检测凋亡相关蛋白监测细胞凋亡。本研究结果表明，在ADR刺激的足细胞中，miR-199b-5p上调，G蛋白信号调节因子10（RGS10）下调。miR-199b-5p的过表达可抑制RGS10表达并刺激足细胞凋亡，而敲低miR-199b-5p可恢复RGS10水平并改善ADR诱导的足细胞凋亡。此外，RGS10过表达可显著逆转miR-199b-5p过表达的作用。此外，足细胞转染miR-199b-5p可激活AKT/雷帕霉素机制性靶标（mTOR）信号通路，RGS10过表达后该信号通路被阻断。综上所述，本研究表明，miR-199b-5p上调可通过激活AKT/mTOR信号通路抑制RGS10表达，从而促进足细胞凋亡。
