1. Anti-fibrotic effect of 6-bromo-indirubin-3'-oxime (6-BIO) via regulation of activator protein-1 (AP-1) and specificity protein-1 (SP-1) transcription factors in kidney cells.
- Author
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Park JS, Jung IA, Choi HS, Kim DH, Choi HI, Bae EH, Ma SK, and Kim SW
- Subjects
- Animals, Cell Line, Connective Tissue Growth Factor drug effects, Connective Tissue Growth Factor genetics, Enzyme Inhibitors pharmacology, Fibrosis, Glycogen Synthase Kinase 3 beta antagonists & inhibitors, Humans, Kidney Diseases pathology, Kidney Tubules, Proximal pathology, Male, Plasminogen Activator Inhibitor 1 drug effects, Plasminogen Activator Inhibitor 1 genetics, Rats, Rats, Sprague-Dawley, Signal Transduction drug effects, Sp1 Transcription Factor drug effects, Sp1 Transcription Factor genetics, Transcription Factor AP-1 drug effects, Transcription Factor AP-1 genetics, Indoles pharmacology, Kidney Diseases drug therapy, Kidney Tubules, Proximal drug effects, Oximes pharmacology
- Abstract
PAI-1 and CTGF are overexpressed in kidney diseases and cause fibrosis of the lungs, liver, and kidneys. We used a rat model of unilateral ureteral obstruction (UUO) to investigate whether 6-BIO, a glycogen synthase kinase-3β inhibitor, attenuated fibrosis by inhibiting PAI-1 and CTGF in vivo. Additionally, TGFβ-induced cellular fibrosis was observed in vitro using the human kidney proximal tubular epithelial cells (HK-2), and rat interstitial fibroblasts (NRK49F). Expression of fibrosis-related proteins and signaling molecules such as PAI-1, CTGF, TGFβ, αSMA, SMAD, and MAPK were determined in HK-2 and NRK49F cells using immunoblotting. To identify the transcription factors that regulate the expression of PAI-1 and CTGF the promoter activities of AP-1 and SP-1 were analyzed using luciferase assays. Confocal microscopy was used to observe the co-localization of AP-1 and SP-1 to PAI-1 and CTGF. Expression of PAI-1, CTGF, TGFβ, and α-SMA increased in UUO model as well as in TGFβ-treated HK-2 and NRK49F cells. Furthermore, UUO and TGFβ treatment induced the activation of P-SMAD2/3, SMAD4, P-ERK 1/2, P-P38, and P-JNK MAPK signaling pathways. PAI-1, CTGF, AP-1 and SP-1 promoter activity increased in response to TGFβ treatment. However, treatment with 6-BIO decreased the expression of proteins and signaling pathways associated with fibrosis in UUO model as well as in TGFβ-treated HK-2 and NRK49F cells. Moreover, 6-BIO treatment attenuated the expression of PAI-1 and CTGF as well as the promoter activities of AP-1 and SP-1, thereby regulating the SMAD and MAPK signaling pathways, and subsequently exerting anti-fibrotic effects on kidney cells., (Copyright © 2021 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)
- Published
- 2022
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