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嗜酸乳杆菌NCFM处理的肠Caco-2细胞的表达谱分析

[Expression profile analysis of intestinal Caco-2 cells treated with Lactobacillus acidophilus NCFM].

作者信息

Wang Mingna, Zhang Guanghui, Han Xiyan, Yao Liyan, Zhou Yanqiu, Jiang Yujun

机构信息

Key Lab of Dairy Science, Ministry of Education, College of Food Science and Engineering, Northeast Agricultural University, Harbin 150030, China.

出版信息

Wei Sheng Wu Xue Bao. 2009 Sep;49(9):1247-52.

Abstract

OBJECTIVE

Lactobacillus acidophilus NCFM is a probiotic strain that promotes human health. We evaluated the influence of Lactobacillus acidophilus NCFM on the genetic expression patterns in the Caco-2 cells.

METHODS

Caco-2 cells were treated with Lactobacillus acidophilus NCFM for 2h. The total RNA of cells was extracted. Hybridization with Human Genome U133 Plus 2.0 Array was performed. The image scanning and data analysis were performed subsequently. Genes with significant expression changes were selected and analyzed. To support the microarray data, three striking difference genes were studied using Real-time RT PCR.

RESULTS

Microarrays analysis showed that the expression levels of 508 genes were altered as compared with the control 473 of them were up-regulated, and 35 were down-regulated. It was supposed that many genes in Caco-2 cell were induced, so that Lactobacillus acidophilus NCFM could play a part in immune system process, antioxidant activity, biological adhesion, cholesterol absorption and so on. Three striking difference genes CCL2, PTX3 and TNFRSF9 which involved immune system process were validated by Real-time RT PCR. And the results of Real-time RT PCR showed the same expression trend as in microarray.

CONCLUSION

Based on the results of gene expression alterations, the beneficial function of Lactobacillus acidophilus NCFM could be more greatly and deeply understood than ever before, and the mechanism of lactic acid bacteria would be revealed.

摘要

目的

嗜酸乳杆菌NCFM是一种可促进人类健康的益生菌菌株。我们评估了嗜酸乳杆菌NCFM对Caco-2细胞基因表达模式的影响。

方法

用嗜酸乳杆菌NCFM处理Caco-2细胞2小时。提取细胞的总RNA。与人类基因组U133 Plus 2.0芯片进行杂交。随后进行图像扫描和数据分析。选择并分析表达有显著变化的基因。为支持芯片数据,使用实时RT-PCR研究三个显著差异基因。

结果

芯片分析显示,与对照相比,508个基因的表达水平发生了改变,其中473个上调,35个下调。推测Caco-2细胞中的许多基因被诱导,因此嗜酸乳杆菌NCFM可能在免疫系统过程、抗氧化活性、生物粘附、胆固醇吸收等方面发挥作用。通过实时RT-PCR验证了涉及免疫系统过程的三个显著差异基因CCL2、PTX3和TNFRSF9。实时RT-PCR结果显示出与芯片相同的表达趋势。

结论

基于基因表达改变的结果,嗜酸乳杆菌NCFM的有益功能能比以往得到更深入的理解,并且乳酸菌的作用机制将被揭示。

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