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目的 观察重组大鼠肝再生增强因子 (ALR)对肝星状细胞株IG12增殖及细胞外基质合成的影响 ,探讨其抗肝纤维化作用的可能机制。方法 利用DNA重组技术构建大鼠ALR原核表达质粒PGEX2T alr,转导XL 1经IPTG诱导ALR表达 ,用SDS/PAGE纯化获得重组肝再生增强因子(rALR)。利用胶原酶灌注梯密度离心分离肝星状细胞 ,用有限稀释法建立大鼠肝星状细胞株IG12。于体外观察rALR对IG12及大鼠成纤维细胞株WFB增殖及细胞外基质合成的影响。结果 rALR可显著抑制IG12的增殖及其细胞外基质的合成 ;但对大鼠成纤维细胞株WFB的增殖无显著抑制作用 ,能抑制WFB产生细胞外基质。结论 原核细胞产生的rALR具有生物学活性 ,它能抑制大鼠肝肌纤维样细胞IG12的增殖及细胞外基质的产生 ;对鼠成纤维细胞株WFB的增殖无显著抑制作用 ,能抑制WFB产生细胞外基质。
Objective To observe the effects of recombinant human augmenter of liver regeneration (ALR) on the proliferation and extracellular matrix synthesis of hepatic stellate cell line IG12 and to explore its possible mechanism of anti-hepatic fibrosis. Methods The prokaryotic expression plasmid PGEX2T alr of rat ALR was constructed by DNA recombination technology. The expression of ALR was induced by IPTG in XL1. Recombinant rat liver regeneration factor (rALR) was purified by SDS / PAGE. The hepatic stellate cells were isolated by gradient centrifugation with collagenase and rat hepatic stellate cell line IG12 was established by limiting dilution. To observe the effect of rALR on proliferation and extracellular matrix synthesis of IG12 and rat fibroblast cell line WFB in vitro. Results rALR significantly inhibited the proliferation of IG12 and the synthesis of extracellular matrix. However, it did not inhibit the proliferation of WFB fibroblasts and inhibited the production of extracellular matrix of WFB. CONCLUSION: The rALR produced by prokaryotic cells has biological activity, which can inhibit the proliferation and extracellular matrix production of hepatic myofibroblast-like cells in vitro. It has no significant inhibitory effect on the proliferation of WFB fibroblasts and inhibits the extracellular Matrix.