title: Cyclosporine A protects podocytes via stabilization of cofilin-1 expression in the unphosphorylated state.
abstract: Podocyte foot process (FP) is dysregulated in nephrotic syndrome. The effacement of podocyte FPs typically arises following perturbations in the actin cytoskeleton. Recent data suggest that the effects of calcineurin (CaN) inhibitor cyclosporine A (CsA) are independent of its effects on T-cells, and CsA has been identified as stabilizing the actin cytoskeleton through stabilizing synaptopodin in podocytes, and thereby directly reducing proteinuria. Other studies showed that CsA could regulate cofilin-1 directly within tubular epithelial cells. However, whether synaptopodin is the only target of CsA or whether the antiproteinuric role of CsA is played by regulating cofilin-1 in podocytes has not been studied. In the present study, changes in the expression and distribution of nephrin, synaptopodin, cofilin-1 and phosphorylated cofilin-1 (pho-cofilin-1) were detected in both puromycin aminonucleoside (PAN) induced nephrotic rats treated with CsA and cultured podocytes exposed to PAN with/without CsA. Cofilin-1, synaptopodin mRNA was knocked down or combined by siRNA to investigate whether cofilin-1 was critical for the protective effect of CsA and whether the effect of CsA on cofilin-1 was independent of its effect on synaptopodin. We found that CsA reduced proteinuria and repaired FP effacement of PAN-induced nephropathy, restored expression of nephrin, synaptopodin, cofilin-1, pho-cofilin-1 both in vivo and in vitro. CsA also repaired actin cytoskeleton impaired by PAN in vitro. The protective effect of CsA was diminished when cofilin-1 was knocked down compared to negative control. Synaptopodin knocked down had no effect on cofilin-1. The protective effect of CsA decreased significantly when cofilin-1 and synaptopodin were simultaneously knocked down compared to only cofilin-1 knock down. In conclusion, the antiproteinuric effect of CsA is derived from the stabilization of the podocyte actin cytoskeleton by upregulating expression of cofilin-1, which was independent of its effect on synaptopodin.
authors.affiliation: Department of Pediatrics, Peking University First Hospital, Beijing 100034, China.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Pediatrics, Peking University First Hospital, Beijing 100034, China.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Pediatrics, Peking University First Hospital, Beijing 100034, China.
authors.forename: Xuejuan
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Pediatrics, Peking University First Hospital, Beijing 100034, China.
authors.forename: Xuejing
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Pediatrics, Peking University First Hospital, Beijing 100034, China.
authors.forename: Suxia
authors.initials: S
authors.lastname: Wang
authors.affiliation: Department of Pediatrics, Peking University First Hospital, Beijing 100034, China djnc_5855@126.com.
authors.forename: Jie
authors.initials: J
authors.lastname: Ding
keywords: Proteinuria; cofilin-1; cyclosporine A