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目的观察重组肝再生增强因子(rALR)、丹参及苦参对大鼠纤维细胞增殖及细胞外基质(ECM)合成的影响.方法利用 DNA 重组技术构建大鼠肝再生增强因子原核表达质粒 PGEX2T-alr,转导 XL-1,IPTG 诱导肝再生增强因子蛋白表达,用 SDS/PAGE 纯化获得 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.于体外观察了 rALR、丹参及苦参对 IG12及大鼠成纤维细胞株 WFB 增殖及 ECM 合成的影响.结果用酸性磷酸酶法测定细胞增殖表明 rALR、丹参及苦参均可显著抑制 IG12的增殖(培养48h,GST 组A:2.207±0.234,苦参组A:1.797±0.158,丹参组A:1.808±0.176,rALR组A:1.779±0.294),丹参及苦参可显著抑制 WFB 的增殖而rALR 对 WFB 的增殖无明显影响(培养72h,GST 组 A:0.940±0.030,苦参组 A:0.797±0.036,丹参组 A:0.821±0.052,rALR 组 A:0.897±0.050).rALR、丹参及苦参可不同程度地抑制 IG12、WFB 合成细胞外基质,但以 rALR 的抑制作用较好.结论原核细胞产生的 rALR 具有生物学活性,rALR、丹参及苦参对 IG12及 WFB 的增殖及 ECM 合成具有不同程度的抑制作用,最佳的联合用药方式有待进一步研究.
Objective To observe the effects of rALR, Salvia miltiorrhiza and Sophora flavescens on the proliferation and extracellular matrix (ECM) synthesis of rat fibroblasts.Methods The prokaryotic expression plasmid PGEX2T-alr , Transduced XL-1 and induced by IPTG, and purified them by SDS / PAGE to obtain rALR.The hepatic stellate cells were isolated by ladder density centrifugation with collagenase and the rat hepatic stellate cell line IG12 The effects of rALR, Salvia miltiorrhiza Bge and Sophora flavescens on the proliferation and ECM synthesis of IG12 and rat fibroblast cell line WFB were observed in vitro.Results The proliferation of ECF was observed by acid phosphatase assay.The results showed that rALR, Salvia miltiorrhiza, (P <0.01), proliferation (cultured for 48h, GST group A: 2.207 ± 0.234, Sophora group A: 1.797 ± 0.158, Salvia group A: 1.808 ± 0.176, rALR group A: 1.779 ± 0.294). Salvia miltiorrhiza and Sophora flavescens could significantly inhibit the proliferation of WFB rALR had no significant effect on the proliferation of WFB (cultured 72h, group A: 0.940 ± 0.030, group A: 0.797 ± 0.036, group A: 0.821 ± 0.052, group A: 0.897 ± 0.050) Sophora flavescens can inhibit IG12, WFB synthesis to varying degrees Extracellular matrix, but the inhibitory effect of rALR is better.Conclusion The prokaryotic cells produce rALR with biological activity, rALR, Salvia miltiorrhiza and Sophora flavescens can inhibit the proliferation and ECM synthesis of IG12 and WFB to varying degrees, the best combination Medication needs further study.