维甲酸核内受体β基因转染对大鼠肝星状细胞增殖和表型的影响

Effect of RARβ transfection on the proliferation and phenotype of rat hepatic stellate cells

目的 研究维甲酸核内受体β（RAR β）转染血小板衍生生长因子（PDGF）激活的HSC后给予相应配体全反式维甲酸（ATRA）对细胞增殖、表型的影响。方法 质粒pCMV-script-RARβ转染经PDGF激活的大鼠HSC, western blot鉴定转染成功后,MTT法、BrdU掺入检测细胞生长情况,免疫细胞化学法检测α-SMA、肌间线蛋白的表达,western blot检测RARβ蛋白表达。结果 受体转染后再给予相应配体可使PDGF激活后HSC的RARβ受体表达升高至少维持144 h,并使其增殖减慢,α-SMA、肌问线蛋白表达减弱,较空载组、PDGF组、未给予配体组、无关配体组差别均有显著性意义。结论 RAR β受体基因转染并给予ATRA可使激活的HSC增殖减慢、表型逆转。

Objective To study the effect of RARβ transfection plus treatment with the corresponding ligand ATRA on the proliferation and phenotype of platelet-derived growth factor (PDGF)-activated hepatic stellate cells (HSC) . Methods PDGF-activated hepatic stellate cells of rats were transfected with eukaryotic expression vector pCMV-script-RAR β, which was verified by western blot. The proliferation of transfected HSC was assayed by BrdU incorporation as well as MTT methods. Their phenotype (α -SMA and desmin) was observed by immunocytochemistry assay with image analysis and RAR β protein expression was detected by western blot. Results Transfection of RAR β gene and treatment with ligand ATRA could increase the expression of RAR β protein for at least 144 h and inhibit the proliferation and the expression of α-SMA and desmin in PDGF-activated HSC. Significant statistical differences were perceived comparing with sham-transfected, only-PDGF treated, non-ligand treated and irrelevant ligand-treated HSC. Conclusions Transfected with RARβ gene as well as using related ligand ATRA could suppress the proliferation and reverse the activation phenotype of activated HSC.