嗜酸乳杆菌NCFM对Caco-2细胞基因表达谱的影响

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

摘要:

摘要：【目的】嗜酸乳杆菌NCFM作为一株具有良好益生功能的模式菌株，采用基因芯片技术对其作用后的宿主细胞基因的变化情况进行分析，在生物、食品等领域具有较大研究价值。【方法】将嗜酸乳杆菌NCFM和Caco-2细胞共同培养2h，提取Caco-2细胞的总RNA，并将RNA反转录成cDNA，与Human Genome U133 Plus 2.0 Array基因表达谱芯片杂交，杂交后进行图像扫描和数据分析，并采用Real-time RT PCR方法对差异表达的基因进行验证。【结果】采用基因芯片方法检测了Caco-2细胞经嗜酸乳杆菌NCFM作用2h后的基因表达变化，发现差异表达基因为508个，其中有473个基因上调，35个基因下调，初步推测Caco-2细胞能诱导多个基因的表达，以发挥嗜酸乳杆菌NCFM的益生功能。并且经Real-time RT PCR验证，表达差异显著的3个免疫调节相关基因CCL2、PTX3和TNFRSF9的确在嗜酸乳杆菌NCFM作用期间高表达。【结论】以上这些结果促进了对嗜酸乳杆菌NCFM益生功能的认识，也为揭示该乳酸菌的作用机理提供了理论基础。

Abstract:

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.