重组大鼠肝再生增强因子对肝星状细胞株IG12的作用

Effects of recombinant augmenter of liver regeneration protein on hepatic stellate cell line IG12

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 augmenter of liver regeneration (ALR) on the proliferation of he patic stellate cell line IG12 and ECM synthesis. Methods Prokaryotic expression plasmid PGEX 2T/alr was constructe d with DNA recombinant technique, the PGEX 2T-alr plasmid was transformed into E .c oli ML-1, and recombinant ALR(rALR) was induced with IPTG and purified with SDS /PAG E. Hepatic stellate cell was isolated with collagenase perfusion and ladder dens ity centrifuge. Hepatic stellate cell line IG12 was cloned with limited diluted method. The effects of rALR on hepatic stellate cell line IG12 and rat fibroblas t cell line WFB were studied. Results The rALR inhibits the prolif eration of IG12 but not WFB. The rALR can inhibits the production of hyaluronic acid,laminin,PⅢP,P ⅠP, and Ⅳ-C of both IG1 2 and WFB. Conclusion rALR is biologically active, and its effects are cell specific . It can inhibit the proliferation and the production of extracellular matrix of hepatic stellate cell line IG12. The rALR can inhibit the production of extrace llular matrix, but not the proliferation of WFB.