硫酸吲哚酚(IS)是一种公认的结合蛋白质的肠源性尿毒症毒素，在慢性肾脏病患者体内蓄积可导致和促进慢性肾脏病的发生和发展。本实验探索了IS是否能引起人肾小球足细胞骨架的损伤及其可能的机理。

以体外培养的人肾小球足细胞系作为研究对象。采用台盼蓝拒染法、MTT法检测足细胞活力；采用免疫荧光法检测足细胞骨架纤维状肌动蛋白(F－actin)、骨架蛋白Synaptopodin改变；Western印迹法分析骨架蛋白Synaptopodin蛋白水平；RT－qPCR法检测Synaptopodin mRNA表达；琼脂糖凝胶电泳法分析足细胞内蛋白激酶A(PKA)活性。

IS使足细胞F－actin、Synaptopodin蛋白荧光减弱；IS还下调Synaptopodin的mRNA (P<0.01)。IS刺激使PKA磷酸化增多；PKA通路阻断剂H89可以上调Synaptopodin的蛋白及mRNA表达(P＝0.002)。

IS可下调足细胞骨架F－actin、Synaptopodin表达，PKA信号通路可能部分参与了Synaptopodin表达的调节，本研究结果为防治和干预CKD发生发展提供了新的思路和潜在的治疗靶点。

Indoxyl sulfate (IS) is a recognized protein-binding enterogenous uremic toxin that accumulates in patients with chronic kidney disease (CKD) and can cause and promote the occurrence and development of CKD. This experiment explored whether IS could cause damage to human glomerular podocyte cytoskeleton and its possible mechanisms in vitro.

The human glomerular podocyte cell line cultured in vitro was taken as the research object. The cell viability was examined by trypan blue exclusion and MTT assay. Immunofluorescence was used to detect changes in podocyte cytoskeletal fibrous-actin (F-actin) and synaptopodin; Western blot was used to analyze the protein level of synaptopodin; RT-qPCR was used to detect the mRNA expression of synaptopodin; Agarose gel electrophoresis was used to analyze the protein kinase A (PKA) activity.

Fluorescence experiments showed that IS reduced the F-actin and synaptopodin protein expression of podocytes. IS also down-regulated the expression of synaptopodin mRNA (P<0.01). IS treatment increased PKA phosphorylation. PKA pathway blocker H89 upregulated the synaptopodin expression of both protein and mRNA (P<0.01).

IS could down-regulate the podocyte cytoskeleton expression of F-actin and synaptopodin; The PKA signaling pathway may partly participate in the regulatory mechanism of synaptopodin expression; The results of this study might provide new ideas and potential therapeutic targets for the prevention and intervention of CKD.