Arecoline-induced pro-fibrotic proteins in LLC-PK1 cells are dependent on c-Jun N-terminal kinase. (17th February 2016)

• Record Type: Journal Article
• Authors: Lin, Sheng-Hsuan; Chiou, Shean-Jaw; Ho, Wan-Ting; Chuang, Chao-Tang; Chuang, Lea-Yea; Guh, Jinn-Yuh
• Is Part Of: Toxicology. Volume 344/346(2016)
• Page Start: 53
• Page End: 60
• Publication Date: 2016-02-17
• Subjects: CKD chronic kidney disease -- TGF-β transforming growth factor-β -- JNK c-Jun N-terminal kinase -- PAI1 plasminogen activator inhibitor-1 -- COX2 cyclooxygenase 2 -- AN areca nut -- ERK extracellular signal-regulated kinase -- DMSO dimethylsulfoxide -- MTT 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium Bromide -- SEM standard errors of the mean; Arecoline -- Proximal tubule -- Renal fibrosis -- TGF-β -- JNK; Toxicology -- Periodicals; Chemicals -- Physiological effect -- Periodicals; 615.9005
• DOI: 10.1016/j.tox.2016.02.004
• Languages: English
• ISSNs: 0300-483X

Abstract

Abstract: Areca nut (AN) chewing is associated with chronic kidney disease (CKD). However, the molecular mechanisms of AN-induced CKD are not known. Thus, we studied the effects of arecoline, a major alkaloid of AN, on proximal tubule (LLC-PK1) cells in terms of cytotoxicity, fibrosis, transforming growth factor-β (TGF-β) and c-Jun N-terminal kinase (JNK). We found that arecoline dose (0.1–0.5 mM) and time (24–72 h)-dependently induced cytotoxicity without causing cell death. Arecoline (0.25 mM) also time-dependently (24–72 h) increased fibronectin and plasminogen activator inhibitor-1 (PAI1) protein expressions. Arecoline (0.25 mM) time-dependently (24–72 h) increased TGF-β gene transcriptional activity and supernatant levels of active TGF-β1. Moreover, arecoline (0.25 mM) activated JNK while SP600125 (a JNK inhibitor) attenuated arecoline-induced TGF-β gene transcriptional activity. SP600125, but not SB431542 (a TGF-β receptor type I kinase inhibitor), attenuated arecoline-induced fibronectin and PAI1 protein expressions. Finally, tubulointerstitial fibrosis occurred and renal cortical expressions of fibronectin and PAI1 proteins increased in arecoline-fed mice at 24 weeks. We concluded that arecoline induced tubulointerstitial fibrosis in mice while arecoline-induced TGF-β and pro-fibrotic proteins (fibronectin, PAI1) are dependent on JNK in LLC-PK1 cells.