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The activation of rat hepatic stellate cells stimulated with hepatitis B virus in vitro
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摘要: 目的观察乙型肝炎病毒(HBV)对大鼠肝星状细胞(HSC)是否有作用。方法用Friedman方法分离大鼠HSC,蔗糖梯度浓度法纯化HBV。用不同浓度HBV刺激HSC 24 h,检测上清液中的前胶原蛋白Ⅲ。选前胶原蛋白Ⅲ分泌增加的细胞作为研究对象,检测其细胞内C/EBPs、PPARγ、RAR表达情况,了解HBV是否影响HSC活化通路。结果 HBV可以呈浓度依赖方式刺激大鼠HSCs分泌前胶原蛋白Ⅲ,在浓度为3.0×105拷贝/ml时可以使前胶原蛋白Ⅲ分泌明显增加。C/EBPs的表达明显减少,而RAR受体表达明显增加、PPARγ表达无变化。结论 HBV对大鼠HSC具有直接激活作用,促其合成Ⅲ型前胶原蛋白,这可能与其刺激C/EBP表达减少和RAR表达增加有关。Abstract: Objective To observe the effects of HBV on rat hepatic stellate cells (HSC) .Methods HSC were isolated by the method of Friedman and HBV was purified by concentrated sucrose gradient.HSC were stimulated with different concentrations of HBV for 24 hrs and the supernatant procollagen Ⅲ was found to be detected.Cells with increased procollagen Ⅲ secretion were selected to check for the intracellular C/EBPs, PPARγ, RAR expression to investigate if HBV had affected the HSC activation pathway.Results Secretion of procollagen III by HBV stimulated rat HSC was concentration dependent.At the concentration of 3.0 × 105 copies/ml, secretion of procollagen Ⅲ increased significantly.The expression of C/EBPs decreased significantly, while the RAR receptor expression was increased and PPARγ remained unchange.Conclusion HBV has a direct stimulus on HSC to promote the synthesis of procollagen Ⅲ, which may be associated with C/EBP decrease and RAR increase.
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Key words:
- hepatitis B virus /
- hepatocytes
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