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以超速离心技术制备大鼠肝刺激因子。采用无血清培养技术培养大鼠肝细胞并观察肝刺激因子作用肝细胞后不同时刻细胞表皮生长因子(epidermalgrowthfactor,EGF)受体mRNA表达的变化,以探讨该因子作用的分子机制。结果显示:肝刺激因子可诱导EGF受体mRNA的表达升高,在给药后2~6小时达高峰。肝刺激因子与EGF合用可明显刺激受体mRNA的表达。Northern杂交分析结果表明,受体mRNA表达增加主要是三种转录本形式中的两种,即5.6和10.0kb所造成。提示该因子可在转录水平调节EGF受体活性。
Rat liver stimulator was prepared by ultracentrifugation. The rat hepatocytes were cultured with serum-free culture and the changes of epidermal growth factor (EGF) receptor mRNA expression at different time points after hepatic stimulation were observed to explore the molecular mechanism of this factor. The results showed that the expression of EGF receptor mRNA was induced by hepatic stimulating factor and peaked at 2-6 hours after administration. Hepatic stimulating factor combined with EGF can obviously stimulate the expression of receptor mRNA. Northern blot analysis indicated that increased receptor mRNA expression was mainly caused by two of the three transcript forms, 5.6 and 10.0 kb. Suggesting that this factor may regulate EGF receptor activity at the transcriptional level.