Yu Shengyou, Zhu Weixue, Yu Li
Department of Pediatrics, Guangzhou First People's Hospital, Guangzhou Medical University, Guangzhou, 510180, Guangdong Province, China.
Department of Pediatrics, Guangzhou First People's Hospital, School of Medicine, South China University of Technology, Guangzhou, Guangdong Province, P. R. China.
Open Life Sci. 2024 Sep 9;19(1):20220958. doi: 10.1515/biol-2022-0958. eCollection 2024.
This study aimed to clarify the role of rapamycin in the PINK1/Parkin signaling pathway in mitophagy in podocytes and the role of voltage-dependent anion channel 1 (VDAC1) in the PINK1/Parkin signaling pathway in mouse glomerular podocytes. For this purpose, podocytes were cultured with rapamycin and observed using microscopy. The apoptosis rate of podocytes was detected by flow cytometry. Changes in the mitochondrial membrane potential were measured. The autophagy-related proteins VDAC1, PINK1, Parkin, and LC3 were detected, and mitochondrial autophagosomes were observed via transmission electron microscopy. In the present study, we demonstrated that the number of podocytes treated with rapamycin was significantly reduced. Compared with those in the control group, the apoptosis rate of podocytes and the degree of mitochondrial membrane potential depolarization were significantly higher. We also found the expression levels of VDAC1, PINK1, Parkin, and LC3 were significantly increased. In the rapamycin-treated group, the numbers of swollen mitochondria and mitochondrial autophagosomes were significantly higher. Finally, we showed that rapamycin can upregulate the expression of VDAC1, PINK1, Parkin, and LC3 in glomerular podocytes, which is correlated with mitophagy. VDAC1 is involved in mitophagy and is related to the PINK1/Parkin signaling pathway, serving as an indicator of mitophagy in podocytes.
本研究旨在阐明雷帕霉素在足细胞线粒体自噬的PINK1/Parkin信号通路中的作用，以及电压依赖性阴离子通道1（VDAC1）在小鼠肾小球足细胞PINK1/Parkin信号通路中的作用。为此，用雷帕霉素培养足细胞并通过显微镜观察。采用流式细胞术检测足细胞凋亡率。测量线粒体膜电位的变化。检测自噬相关蛋白VDAC1、PINK1、Parkin和LC3，并通过透射电子显微镜观察线粒体自噬体。在本研究中，我们证明用雷帕霉素处理的足细胞数量显著减少。与对照组相比，足细胞凋亡率和线粒体膜电位去极化程度显著更高。我们还发现VDAC1、PINK1、Parkin和LC3的表达水平显著增加。在雷帕霉素处理组中，肿胀线粒体和线粒体自噬体的数量显著更多。最后，我们表明雷帕霉素可上调肾小球足细胞中VDAC1、PINK1、Parkin和LC3的表达，这与线粒体自噬相关。VDAC1参与线粒体自噬并与PINK1/Parkin信号通路相关，是足细胞线粒体自噬的一个指标。
