Children's Hospital, CO, United States.
Children's Hospital, CO, United States.
Nefrologia (Engl Ed). 2024 Jan-Feb;44(1):90-99. doi: 10.1016/j.nefroe.2023.04.003. Epub 2023 May 5.
Activation of the focal adhesion kinase (FAK) in podocytes is involved in the pathogenesis of minimal change disease (MCD), but the pathway leading to its activation in this disease is unknown. Here, we tested whether podocyte β1 integrin is the upstream modulator of FAK activation and podocyte injury in experimental models of MCD-like injury.
We used lipopolysaccharide (LPS) and MCD sera to induce MCD-like changes in vivo and in cultured human podocytes, respectively. We performed functional studies using specific β1 integrin inhibitors in vivo and in vitro, and integrated histological analysis, western blotting, and immunofluorescence to assess for morphological and molecular changes in podocytes. By ELISA, we measured serum LPS levels in 35 children with MCD or presumed MCD (idiopathic nephrotic syndrome [INS]) and in 18 healthy controls.
LPS-injected mice showed morphological (foot process effacement, and normal appearing glomeruli on light microscopy) and molecular features (synaptopodin loss, nephrin mislocalization, FAK phosphorylation) characteristic of human MCD. Administration of a β1 integrin inhibitor to mice abrogated FAK phosphorylation, and ameliorated proteinuria and podocyte injury following LPS. Children with MCD/INS in relapse had higher serum LPS levels than controls. In cultured human podocytes, β1 integrin blockade prevented cytoskeletal rearrangements following exposure to MCD sera in relapse.
Podocyte β1 integrin activation is an upstream mediator of FAK phosphorylation and podocyte injury in models of MCD-like injury.
足细胞中粘着斑激酶(FAK)的激活参与了微小病变病(MCD)的发病机制,但导致其在该疾病中激活的途径尚不清楚。在这里,我们测试了足细胞β1 整合素是否是 FAK 激活和 MCD 样损伤实验模型中足细胞损伤的上游调节剂。
我们分别使用脂多糖(LPS)和 MCD 血清在体内和体外诱导 MCD 样改变。我们在体内和体外使用特定的β1 整合素抑制剂进行功能研究,并进行组织学分析、Western blot 和免疫荧光,以评估足细胞的形态和分子变化。通过 ELISA,我们测量了 35 名 MCD 或疑似 MCD(特发性肾病综合征 [INS])患儿和 18 名健康对照者的血清 LPS 水平。
注射 LPS 的小鼠表现出形态学(足突融合,光镜下肾小球正常)和分子特征(突触蛋白丢失,nephrin 定位错误,FAK 磷酸化),这些特征与人 MCD 相似。给予小鼠β1 整合素抑制剂可阻断 FAK 磷酸化,并改善 LPS 后蛋白尿和足细胞损伤。复发的 MCD/INS 患儿的血清 LPS 水平高于对照组。在体外培养的人足细胞中,β1 整合素阻断可防止 MCD 血清暴露后细胞骨架的重排。
在 MCD 样损伤模型中,足细胞β1 整合素的激活是 FAK 磷酸化和足细胞损伤的上游介质。