摘要
目的观察慢病毒介导的BMP-7高表达是否能抑制大鼠HSC-T6细胞发生上皮-间叶转化(epithelial to mesenchymal tran-sition,EMT),并能拮抗TGF-β1的促EMT作用。方法构建大鼠BMP-7慢病毒表达载体,体外感染HSC-T6细胞株,将成功感染BMP-7重组慢病毒的细胞给予TGF-β1(5 ng/mL)或溶酶体处理。Real-time PCR检测α-SMA、S100A4、E-cadherin mRNA转录水平。Western blotting检测α-SMA、S100A4、E-cadherin、Ⅰ型胶原蛋白表达水平。结果 BMP-7慢病毒感染HSC-T6后,无论TGF-β1存在与否,real-time PCR检测到α-SMA、S100A4 mRNA转录下调,E-cadherin转录上调。Western blotting显示α-SMA、S100A4及Ⅰ型胶原蛋白表达下调,E-cadherin蛋白表达上调。实验组与空病毒对照组比较,差异有统计学意义(P<0.05),空病毒对照组与空白组比较,差异无统计学意义。结论 BMP-7能有效拮抗TGF-β1对HSC-T6的促EMT作用,可能成为肝纤维化治疗的新途径。
Objective To observe whether lentivirus-mediated overexpression of BMP-7 inhibits epithelial-mesenchymal transition（EMT） in HSC-T6 with or without TGF-β1 treatment.Methods Lentiviral vectors overexpressing BMP-7 were constructed and transfected into HSC-T6 cells.Transfected cells were incubated with or without TGF-β1（5 ng/mL） treatment for another 48 h.The mRNAs of α-smooth muscle actin（α-SMA）,S100A4 and E-cadherin were analyzed by real-time PCR.The protein of α-SMA,S100A4,E-cadherin and collagenⅠ were determined by western blotting.Results HSCs were successfully infected.Real-time PCR results showed that the expression of E-cadherin was increased while α-SMA and S100A4 were decreased with or without TGF-β1（5 ng/mL） treatment.Similiary,western blotting revealed that overexpression of BMP-7 supressed α-SMA,S100A4 and collagen I expression while enhanced E-cadherin expression in HSC with or without TGF-β1 treatment.Conclusion These data demonstrate that BMP-7 can effectively antagonize TGF-β1 induced EMT,which may provide new therapeutic strategy for liver fibrosis.
出处
《胃肠病学和肝病学杂志》
CAS
2011年第2期128-131,共4页
Chinese Journal of Gastroenterology and Hepatology
基金
国家自然科学基金资助项目(30971332)
上海市浦江人才计划项目(10PJ1407600)
关键词
骨形态发生蛋白-7
肝星状细胞
上皮-间叶转化
Bone morphogenic protein-7
Hepatic stellate cell
Epithelial to mesenchymal transition
