甘草甜素诱导的大鼠肝星状细胞差异表达基因检测

Detection of differentially expressed genes in rat hepatic stellate cells treated with glycyrrhizin

目的:检测甘草甜素诱导的大鼠肝星状细胞的差异表达基因,为探讨甘草甜素抗肝纤维化的作用机制提供分子生物学依据。方法:大鼠肝星状细胞分2组培养,实验组在培养液中加入甘草甜素,终浓度达1.0mmol/L;对照组不作处理。继续培养24h,应用抑制性消减杂交技术构建甘草甜素诱导的肝星状细胞差异表达基因的cDNA文库,将杂交产物与pGEM T载体连接,转化感受态DH5α,阳性克隆用PCR技术扩增后进行测序及同源性分析。结果:成功构建了甘草甜素诱导的肝星状细胞cDNA消减文库。测序分析结果表明,在cDNA消减文库中包含3个铁蛋白重链基因的阳性克隆。结论:甘草甜素能诱导肝星状细胞铁蛋白重链的表达。调控铁蛋白重链基因的表达可能是甘草甜素抗肝纤维化作用的分子生物学机制之一。

Aim： To detect the differentially expressed genes in rat hepatic stellate cells treated with glycyrrhizin. Mothods： Rat hepatic stellate cells were treated with glycyrrhizin and the subtractive cDNA library from hepatic stellate cell was constructed by suppression subtractive hybridization （SSH）. The cDNAs was sequenced and analyzed in GenBank with Blast search after colony P.CR. Results ： The suhtractive cDNAs library of genes up-regulated in hepatic stellate cells treated with glycyrrhizin was constructed successfully. Sequencing analysis showed that there were three positive clones containing ferritin heavy chain gene. Conclusion： Glycyrrhizin could up-regulate the expression of ferritin heavy chain gene in hepatic stellate cells,which may contribute to the anti-fibrosis of glycyrrhizin.