| 张岩,张亮,周一鸣,赵艳君,果德安,周玉祥,程京.用酿酒酵母全基因组DNA芯片研究盐酸小檗碱的药理作用机制[J].中国中西医结合杂志,2003,(1):48-53 |
| 用酿酒酵母全基因组DNA芯片研究盐酸小檗碱的药理作用机制 |
| Study on Pharmacological Action Mechanism of Berberine Chloride by Genome-wide Expression Profiling of Saccharomyces cerevisiae |
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| DOI： |
| 中文关键词: 酿酒酵母 DNA芯片 盐酸小檗碱 基因差异表达 |
| 英文关键词:Saccharomyces cerevisiae DNA microarray berberine chloride gene differential expression |
| 基金项目:国家自然科学基金资助项目 (No .398890 0 1 ;No .3982 51 0 8;No.30 0 70 91 7) ;国家重点基础研究资助项目 (G1 9990 1 1 6) |
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| 中文摘要: |
| 目的 :利用自己构建的酿酒酵母全基因组DNA芯片 ,分析盐酸小檗碱处理酵母细胞后对其整个基因表达谱的影响 ,在基因水平上分析盐酸小檗碱的药理机制。方法 :用盐酸小檗碱 (10 0 μg/ml)处理酵母细胞 90min后 ,抽提细胞的RNA进行反转录荧光标记cDNA ,与DNA芯片进行杂交。用激光扫描仪检测杂交信号。结果 :共有 5 45个基因〔占全部基因 (6 183个 )的 9%〕受到诱导 ,38个基因 (占全部基因的 0 6 % )受到抑制。被诱导的基因包括与细胞生长、蛋白质合成及应激反应相关的基因。受抑制的基因包括与氧化磷酸化过程相关的基因和好氧基因。与金属离子平衡 ,特别是与铁离子和铜离子平衡有关的基因表达有较大变化。结论 :处理 90min后 ,酵母细胞在转录水平上表现出对盐酸小檗碱处理的适应性。盐酸小檗碱的药理作用可能和引起细胞内的氧浓度变化及氧化还原反应产生的自由基有关。细胞容易对盐酸小檗碱产生抗药性的原因是由于与细胞抗药性相关的基因受到了明显的诱导。 |
| 英文摘要: |
| Objective: To analyse the pharmacological action mechanism of berberine chloride at the molecular level by using the genome wide Saccharomyces cerevisiae DNA microarray constructed in our laboratory. Methods: S cerevisiae was treated with 100 μg/ml berberine chloride for 90 min. RNA was extracted and reverse transcribed to cDNA labeled with fluorescent dye. cDNA was hybridized with the DNA microarray and hybridization signals were detected with laser scanner. Results: Altogether 547 genes (9.2% of all the genes) were up regulated and 38 genes (0.64% of all the genes) were down regulated more than 2 fold. Genes involved in cell growth, protein synthesis and stress response exhibited obvious induction. In the meanwhile, it was also noticed that 15 genes related to oxidative phosphorylation were repressed apart from the aerobic genes. There were obvious expression alterations in the genes involved in ion homeostasis especially in iron and copper. Conclusion: Yeast cell had developed adaptive responses to the berberine brought stress after treatment for 90 min. The pharmacological action mechanism of the berberine were possibly related to the intracellular change of oxygen concentration and the free radicals produced in metal ion participated redox reaction. In addition, the gene expression level of the four pleiortropic drug resistance genes PDR5, PDR15, YOR1 and SNQ2 was increased, which might account for the cellular drug resistance to berberine. |
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