论文部分内容阅读
目的研究维甲酸核内受体RXRα基因转染血小板趋化生长因子(PDGF)激活肝星状细胞(HSC)后给予相应配体9-cis-RA对细胞增殖和表型的影响.方法真核表达质粒pcDNA3.1-RXRα转染经血小板趋化生长因子激活的大鼠肝星状细胞,经Western blot鉴定转染成功后用MTT法、BrdU掺入检测细胞生长情况,并用免疫细胞化学法加图像分析检测α-SMA和desmin 的表达.结果受体转染后再给予相应配体可使血小板趋化生长因子激活的肝星状细胞RXRα受体表达升高至少维持168小时并使其增殖减慢;α-SMA及desmin表达减弱,较空载组、PDGF组、未给予配体组、无关配体组的差别均有显著性意义.结论 RXRα受体基因转染并结合使用相应配体9-cis-RA可使激活的肝星状细胞增殖减慢、表型逆转.“,”Objective To study the effect of retinoid X receptor alpha (RXRα) transfection plus treatment with the RXRα ligand, 9-cis-RA, on the proliferation and phenotype of platelet-derived growth factor (PDGF)-activated hepatic stellate cells (HSCs). Methods PDGF activated rat hepatic stellate cells were transfected with eukaryotic expression vector pcDNA3.1- human RXRα, and confirmed by Western blot. Proliferation of transfected HSC was assayed by bromodeoxyuridine (BrdU) incorporation as well as MTT, and the phenotype (α-smooth muscle actin, desmin) was observed by immunocytochemistry with image analysis. Results Transfection of the RXRα gene and treatment with ligand 9-cis-RA of PDGF-activated HSCs extended the increased expression of RXRα protein for at least 168 hours. Cell proliferation and expressions of alpha-smooth muscle actin (α-SMA) and desmin were blocked, compared with groups of sham-transfected, PDGF-activated, no transfection, no ligand treatment, and irrelevant ligand treated HSCs. Conclusion Transfection with the RXRα gene followed by 9-cis-RA ligand treatment will inhibit the proliferation and reverse the phenotype of activated HSC.