肝星状细胞HSC-T6体外肝纤维化模型的建立
Establishment of a liver fibrosis model in vitro by hepatic stellate HSC-T6 cells
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摘要: 目的 研究以肝星状细胞HSC-T6为靶细胞,建立体外肝纤维化模型.方法 小鼠腹腔巨噬细胞先后用卡西霉素和脂多糖刺激培养24h制备巨噬细胞条件培养基.肝星状细胞增殖和胶原合成分别采用结晶紫染色法和3H-脯氨酸掺入法测定.结果 血清和巨噬细胞条件培养基可显著促进HSC-T6细胞增殖与胶原合成.IL-1、TNF、EGF、FGF和PDGF均可促进HSC-T6细胞增殖,其中PDGF的促增殖能力最强.TGFβ1可剂量依赖地促进HSC-T6细胞胶原的合成.结论 用血清、巨噬细胞条件培养基、PDGF或TGFβ刺激HSC-T6细胞增殖和胶原合成作为体外肝纤维化模型是可行的.Abstract: Objective To establish a liver fibrosis model in vitro by hepatic stellate HSC-T6 cells. Methods Mouse peritoneal macrophages were primed with calcimycin 10-6mol/L for 8h then elicited by lipopolysaccharides(LPS) 100μg/L for 6h to prepare macrophage conditioned medium(MCM). Proliferative activity and collagen stimulating activity was determined by crystal violet staining assay and[3H]-proline incorporation assay using rat hepatic stellate HSC-T6 cell. Results Serum(0%-20%) and MCM(1:32-1:2) concentration-dependently enhanced HSC-T6 cell proliferation and collagen synthesis. Among IL-1, TNF, EGF, FGF and PDGF, PDGF showed the highest proliferation enhancing activity. TGFβ1 increased HSC-T6 cell collagen synthetic capacity. Conclusion It is feasible to establish an in vitro hepatic fibrosis model selecting HSC-T6 cell proliferation and collagen synthesis as indexes with stimulating factors serum, MCM and cytokines.
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Key words:
- hepatic stellate cells /
- macrophages /
- cytokine /
- fibrosis
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