【摘要】 目的 :研究莪术油对HSC T6细胞基因表达的影响。方法 :从基因库中查询 5 0种肝纤维化相关基因的mRNA序列 ,用寡核苷酸探针设计软件设计探针 ,在PE890 9DNA合成仪上合成寡核苷酸 ,用OGR 0 4点样仪及醛基化玻片制备成基因芯片。以莪术油不同浓度含药培养液培养HSC T6细胞。根据细胞毒性实验 ,确定细胞存活率在5 0 %以上的药物浓度作为实验所用浓度 ,每组设空白对照 ,分别按 1小时、 6小时、 12小时、 2 4小时 4个时间段收集细胞 ,按操作步骤提取细胞总RNA ,经反转录荧光标记 ,杂交和洗涤 ,用Genepix 40 0 0B扫描仪扫描芯片 ,ImaGene4 2软件进行数据分析和归一化处理 ,使用看家基因和阳性对照对Cy3和Cy5扫描结果进行校正。 结果 :莪术油78 12 5 μg/ml作用HSC T6细胞 2 4小时 ,可使基因TIMP 2、IL 6表达分别下调 2 3、 2 2倍。 结论 :从分子水平揭示了莪术有效成分莪术油的抗肝纤维化机制。更多还原

【Abstract】 Objective: To investigate the effects of curcuma aramatica oil on the gene expression in cultured activated hepatic stellate cells. Methods:The mRNA sequence of 50 liver fibrosis related genes was selected from gene bank, loigonucleotide probes were designed by software of designing oligonucleotide probe.The oligonucleotides were synthesized in PE8909DNA synthesizing instrument, and the oligonucleotide microarray was prepared with OGR 04 droping instrument and aldehyded glass chip.Cultured HSC T6 cells were treated with curcuma aramatica oil. According to the experiment of cells toxicity, took the appropriate concentrations of medicines whose cells livability was higher than 50% as experiment concentrations. The blank comparison each team in the same time was setted up, and gathered the cells at 4 point of 1h、6h、12h、24h, respectively. The total RNA with trizol reagent according to the operation process was distilled, then labeled cDNAs from total RNA with cy3 dUTP and cy5 dUTP.These labeled cDNAs were hybrided to an oligonucleotide microarray, then washed the oligonucleotide microarray a few times. Afterwards, scanned the gene microarray by scanner Genepix 4000B, and analysed difference gene expression of HSC T6 cells with ImaGene 4 2 software. Results:After HSC T6 cells were cultured in medium with curcuma aramatica oil(CAO) 78 125μg/ml for 24h,gene expression of tissue inhibitor of metalloprotenase1(TIMP 1)was down regulated 2 2 folds, TIMP 2 and Interleukin 6(IL 6) were down regulated 2 3、2 2 folds, respectively. Conclusion:These results not only show the possible molecular mechanism of anti hepatic fibrosis of CAO which is component of Ezhu,but also the possibility of TCM research by DNA microarray.更多还原