双环醇对刀豆蛋白A引起肝损伤小鼠肝脏基因表达谱的影响

Effect of bicyclol on gene expression profiles in mice with liver injury induced by concanavalin A

本实验研究双环醇对刀豆蛋白A(concanavalin A,Con A)静脉注射引起免疫性肝损伤小鼠肝脏基因表达谱变化的影响,探讨双环醇肝保护作用的分子机制。小鼠于注射Con A26.5 mg.kg-1前24、8及1 h分别口服双环醇250 mg.kg-1。测定血清丙氨酸转氨酶(alanine aminotransferase,ALT)及天冬氨酸转氨酶(aspartateaminotransferase,AST)水平,提取小鼠肝脏总RNA,经反转录用Cy3-dUTP和Cy5-dUTP分别标记制备cDNA探针。将cDNA探针与BiostarM-40S小鼠基因表达谱芯片进行杂交,经ScanArray 4000扫描仪扫描芯片并用GenePix Pro 3.0软件进行分析。双环醇可显著抑制刀豆蛋白A引起的血清ALT和AST升高。与刀豆蛋白A对照组相比,双环醇给药组有287条基因发生差异表达,占芯片基因总数的7.00%。其中166条基因表达量明显下调,121条基因表达量明显上调。表达变化的基因主要涉及代谢与细胞色素P450、应激与炎症凋亡、细胞周期调控、信号传导以及再生等相关功能。双环醇对刀豆蛋白A引起小鼠肝损伤肝脏基因表达谱变化具有一定的影响,此结果对今后深入研究双环醇的肝脏保护作用特点和临床应用具有重要意义。

The aim of this study was to investigate the effect of the novel antihepatitis drug bicyclol on the gene expression profiles in concanavalin A （Con A） intoxicated mice by using cDNA microarray analysis. Bicyclol （250 mg·kg^-1） was given orally to mice three doses before Con A intravenous injection （26.5 mg·kg^-1 ）. Serum levels of aminotransferases were examined by biochemical methods. Liver mRNA was extracted and reversely transcribed to cDNA with the incorporation of labeled Cy3-dUTP and CyS-dUTP, separately. The probes were hybridized to the cDNA microarray. The acquired image was scanned and analyzed by Cenepix Pro 3.0 software. Microarray analysis showed that 287 genes exhibited differential expression in bicyclol group, in which 121 genes were up-regulated and 166 genes were down- regulated comparing with that of untreated Con A intoxicated mice. The differential gene expression after bicyclol treatment was involved in the biotransformation, protein synthesis, degradation and circadian rhythm, proliferation and signal transduction. Bicyclol might regulate a series of genes expressions in Con A intoxicated mice.