Department of Nephrology, The Second Xiangya Hospital, Central South University, Changsha, China.
Department of Nephrology, The Third Xiangya Hospital, Central South University, No.138, Tongzipo Road, Changsha, 410013, Hunan Province, China.
Sci Rep. 2023 Jul 8;13(1):11088. doi: 10.1038/s41598-023-37510-5.
To investigate the effects and mechanism of Vitamin D receptor (VDR) signaling on arteriovenous fistula (AVF) endothelial cell injury. Venous tissues of AVF stenosis patients were collected and analyzed, vascular morphology, reactive oxygen species (ROS), and the expression of VDR, P66Shc, fibronectin (FN), collagen-1 (Col-1) were detected. In addition, human umbilical vein endothelial cells (HUVECs) was used in in vitro studies. HUVECs was incubated with transforming growth factor-beta (TGF-β, 50 ng/ml). Aditionally, paricalcitol, VDR overexpression plasmid and Pin1 inhibitor Juglone were used to investigate the regulatory mechanism of VDR in mitochondrial ROS. The parameters of ROS (e.g. MitoSox) and the expression of FN, Col-1 were tested. Moreover, the mitochondrial translocation of P66Shc was analyzed. The expression of VDR was obviously decreased in the venous tissues of AVF stenosis patients. On the contrary, the expression of P66Shc, P-P66Shc, FN, Col-1 and 8-OHdG were increased significantly in the venous tissues of AVF stenosis patients (P < 0.05). In line with this, the level of mitochondrial ROS and the expression of P66Shc, P-P66Shc, FN, Col-1 increased obviously in HUVECs cells under TGF-β condition. Both VDR over-expression plasmid and Pin1 inhibitor Juglone could alleviate TGF-β induced endothelial injury. Mechanistically, VDR overexpression plasmid and Juglone could inhibit the expression of Pin1, and then restrain P66Shc mitochondrial translocation, eventually reduce the level of mitochondrial ROS. Our research indicated that activation of VDR could alleviate venous endothelial cell dysfunction through inhibiting Pin1-mediated mitochondrial translocation of P66Shc and consequently reducing mitochondrial ROS. It suggested that VDR signaling might be an effective target for AVF stenosis treatment.
探讨维生素 D 受体（VDR）信号对动静脉瘘（AVF）内皮细胞损伤的作用及机制。收集并分析 AVF 狭窄患者的静脉组织，检测血管形态、活性氧（ROS）以及 VDR、P66Shc、纤维连接蛋白（FN）、胶原-1（Col-1）的表达。此外，在体外研究中使用人脐静脉内皮细胞（HUVECs）。将 HUVECs 与转化生长因子-β（TGF-β，50ng/ml）孵育。此外，还使用 paricalcitol、过表达 VDR 的质粒和 Pin1 抑制剂 Juglone 来研究 VDR 对线粒体 ROS 的调节机制。测试了 ROS 的参数（例如 MitoSox）以及 FN、Col-1 的表达。此外，还分析了 P66Shc 的线粒体易位。AVF 狭窄患者静脉组织中 VDR 的表达明显降低。相反，AVF 狭窄患者静脉组织中 P66Shc、P-P66Shc、FN、Col-1 和 8-OHdG 的表达明显增加（P < 0.05）。与此一致的是，在 TGF-β 条件下，HUVECs 细胞中线粒体 ROS 水平以及 P66Shc、P-P66Shc、FN、Col-1 的表达明显增加。过表达 VDR 的质粒和 Pin1 抑制剂 Juglone 均可减轻 TGF-β 诱导的内皮损伤。从机制上讲，过表达 VDR 的质粒和 Juglone 可以抑制 Pin1 的表达，从而抑制 P66Shc 的线粒体易位，最终降低线粒体 ROS 水平。我们的研究表明，激活 VDR 可以通过抑制 Pin1 介导的 P66Shc 线粒体易位，从而减少线粒体 ROS，从而减轻静脉内皮细胞功能障碍。这表明 VDR 信号可能是治疗 AVF 狭窄的有效靶点。
