TGF-ß1 induces podocyte injury through Smad3-ERK-NF-κB pathway and Fyn-dependent TRPC6 phosphorylation.
Cell Physiol Biochem
; 26(6): 869-78, 2010.
Article
em En
| MEDLINE
| ID: mdl-21220918
ABSTRACT
TGF-ß1 plays an important role on podocyte injury and glomerular diseases, while the underlying molecular mechanisms are still elusive. Here, the potential role of the ion channel TRPC6 and the proximal signaling was explored in TGF-ß1-treated mouse podocyte. Our results showed that TGF-ß1 significantly increased podocyte apoptosis and induced obvious disorganization of actin filaments in a time-dependent pattern. In TGF-ß1-treated podocyte, TRPC6 protein, especially the phosphorylated TRPC6, and the cytosolic free Ca(2+) level upregulated, which was evidently inhibited by the specific knockdown of TRPC6. TRPC6 knockdown also alleviated TGF-ß1-induced podocyte apoptosis. Moreover, the Src kinase Fyn increased obviously in TGF-ß1-treated podocyte, displaying increment of the active form pY418 and reduction of the inactive form pY530. Immunoprecipitation assay revealed that Fyn interacts with TRPC6 in podocyte. Notably, Fyn knockdown blocked TRPC6 phosphorylation and intracellular Ca(2+) increment following TGF-ß1 stimulation, but not affect the expression of TRPC6 protein. In addition, Western blot showed that TGF-ß1 induced significant activation of p-Smad3, p-ERK and RelA/p65. Importantly, obvious translocation of ERK and RelA/p65 to nuclei was observed in TGF-ß1-treated podocyte, which was reduced by ERK inhibitor U0126. Both U0126 and NF-κB inhibitor PDTC obviously inhibited the increment of TRPC6 protein and the flux of cytosolic free Ca(2+) induced by TGF-ß1. Together, we provide evidences that TGF-ß1 induces podocyte damage by upregulating TRPC6 protein most possibly through Smad3-ERK-NF-κB pathway, in which Fyn-dependent tyrosine phosphorylation of TRPC6 might exert a crucial role on the activation of its channel function.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Sistema de Sinalização das MAP Quinases
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Podócitos
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Proteínas Proto-Oncogênicas c-fyn
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Canais de Cátion TRPC
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Fator de Crescimento Transformador beta1
Limite:
Animals
Idioma:
En
Ano de publicação:
2010
Tipo de documento:
Article