title: Effects of retinoic acid on proliferation, phenotype and expression of cyclin-dependent kinase inhibitors in TGF-beta1-stimulated rat hepatic stellate cells.
abstract: AIM:To study the molecular mechanisms of retinoic acid (RA)on prolix-feration and expression of cyclin-dependent kinase inhibitors (CKI), i.e.p16, p21 and p27 in cultured rat hepatic stellate cells (HSC) stimulated with transforming growth factor beta 1 (TGF-beta1).METHODS:HSC were isolated from healthy rat livers and cultured.After stimulated with 1mg/L TGF-beta1, subcultured HSC were treated with or without 1nmol/L RA. MTT assay, immunocytochemistry (ICC) for p16, p21, p27 and alpha-smooth muscle actin (alpha-SMA) protein, in situ hybridization (ISH) for retinoic acid receptor beta 2 (RAR-beta2) and p16, p21 and p27 mRNA and quantitative image analysis (partially) were performed.RESULTS:inhibited HSC proliferation (41.50%,P<0.05),decreased the protein level of alpha-SMA (55.09%, P<0.05), and induced HSC to express RAR-beta2 mRNA. In addition, RA increased the protein level of p16 (218.75%, P <0.05) and induced p21 protein expression; meanwhile, p27 was undetectable by ICC in both control and RA-treated HSC. However, RA had no influence on the mRNA levels of p16, p21 or p27 as determined by ISH.CONCLUSION:Up-regulation of p16 and p21 on post-transcriptional level may contribute, in part, to RA inhibition of TGF-beta 1-initiated rat HSC activation in vitro.
authors.forename: Guang-Cun
authors.initials: GC
authors.lastname: Huang
authors.forename: Jin-Sheng
authors.initials: JS
authors.lastname: Zhang
authors.forename: Yue-E
authors.initials: YE
authors.lastname: Zhang