Involvement of cytosolic phospholipase A2 alpha signalling pathway in spontaneous and transforming growth factor-beta-induced activation of rat hepatic stellate cells.

BACKGROUND:Hepatic stellate cells (HSCs) are extracellular matrix-producing cells that play a pivotal role in liver fibrogenesis. During liver injury and when cells are placed in vitro, HSCs undergo phenotypic transition from quiescent retinoid-storing cells to activated retinoid-deficient myofibroblast-like cells. Although several mediators including reactive oxygen species, platelet derived growth factor, transforming growth factor-beta (TGF-β) and tumour necrosis factor-alpha (TNF-α) were implicated in HSC activation, the cellular signalling pathways that regulate this process remain incompletely defined. AIMS:The objectives of this study were to evaluate the role of cytosolic phospholipase A(2) alpha (cPLA(2)α) and peroxisome proliferator-activated receptor-beta/delta (PPAR-β/δ) in HSC activation. METHODS:Rat HSCs were isolated, purified, cultured and stimulated with TGF-β1 in the presence or absence of the selective cPLA(2)α inhibitor, arachidonyltrifluoromethyl ketone (AACOCF(3)). The activation status of HSC was evaluated by immunofluorescent staining of alpha-smooth muscle actin (α-SMA) and by measuring the expression of cPLA(2)α, cyclooxygenase 2 (COX-2) and PPAR-β/δ using western blot analysis. RESULTS:Rapid and significant increase in cPLA(2)α expression was observed during activation of HSCs. These events preceded the elevation of PPAR-β/δ and the expression of α-SMA. Elevated expression of cPLA(2)α, but not COX-2, was also observed during TGF-β-induced HSC activation. The TGF-β-induced α-SMA expression was blocked by AACOCF(3). Furthermore, transfection of a cPLA(2)α expression vector enhanced the transcription activity of PPAR-β/δ and the expression of α-SMA in HSCs. CONCLUSION:cPLA(2)α-mediated induction of PPAR-β/δ is a novel intracellular signalling pathway in spontaneous and TGF-β induced activation of HSCs and could be a potential therapeutic target for the treatment of liver fibrosis.

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