Zhu Yanji, Xie Guiling, Qi Fangyan, Tang ShenWei, Xun Wenlong
Department of Pediatrics, People's Hospital of Rizhao, Rizhao 276800, PR China.
College of Clinical Medicine, Jining Medical University, Jining 272000, PR China.
Tissue Cell. 2025 Apr;93:102647. doi: 10.1016/j.tice.2024.102647. Epub 2024 Dec 1.
Podocyte injury is considered one of the main causes of kidney diseases occurrence and development. We have demonstrated that Ecto-5'-Nucleotidase (CD73) upregulated during podocyte injury, yet its function in podocyte is still unclear. Mouse podocytes cell line (MPC5) were exposed to the adriamycin (ADR, 0.25 μg/ml) to establish the model of podocytes injury, as well as low expression CD73 with lentivirus transfected shRNA. CD73 expression was verified by western blot and immunofluorescence assay. Cytokines (IL-1β, IL-18), apoptosis and apoposis-related protein (Bax, Caspase-3, Desmin) levels were measured using ELISA assay, Flow cytometry and Western blot, respectively. CD73, the cytokines of IL-1β and IL-18, apoptosis rate and the expression of Bax, Caspase-3 and Desmin were significantly increased in ADR group compared with the control group. Moreover, we also successfully constructed a CD73 down-expressed podocytes cell line. However, in comparsion with the ADR group, the cytokines of IL-1β and IL-18, apoptosis rate and the expression of Bax, Caspase-3 and Desmin protein were remarkably lowered in the ADR+CD73 shRNA group. These findings demonstrate that CD73 alleviates podocyte damage by reducing the inflammation and increasing apoptosis.
足细胞损伤被认为是肾脏疾病发生和发展的主要原因之一。我们已经证明，外切5'-核苷酸酶（CD73）在足细胞损伤时上调，但其在足细胞中的功能仍不清楚。将小鼠足细胞系（MPC5）暴露于阿霉素（ADR，0.25μg/ml）以建立足细胞损伤模型，并用慢病毒转染的shRNA降低CD73的表达。通过蛋白质免疫印迹和免疫荧光测定法验证CD73的表达。分别使用酶联免疫吸附测定法、流式细胞术和蛋白质免疫印迹法测量细胞因子（IL-1β、IL-18）、细胞凋亡及凋亡相关蛋白（Bax、Caspase-3、结蛋白）的水平。与对照组相比，ADR组中CD73、IL-1β和IL-18细胞因子、细胞凋亡率以及Bax、Caspase-3和结蛋白的表达均显著增加。此外，我们还成功构建了CD73表达下调的足细胞系。然而，与ADR组相比，ADR+CD73 shRNA组中IL-1β和IL-18细胞因子、细胞凋亡率以及Bax、Caspase-3和结蛋白的表达均显著降低。这些发现表明，CD73通过减轻炎症和增加细胞凋亡来减轻足细胞损伤。
