嗜酸乳杆菌NCFM对Caco-2细胞中PTX3基因表达的影响

Expression of PTX3 gene in Caco-2 cells treated with Lactobacillus acidophilus NCFM

摘要:

摘要：【目的】研究嗜酸乳杆菌NCFM对肠道上皮细胞中免疫与炎症介质因子PTX3表达的影响，并进一步揭示其调节机制。【方法】嗜酸乳杆菌NCFM与Caco-2细胞共培养0、2、4、8和12 h，提取细胞RNA，采用Real Time RT-PCR方法检测PTX3基因的表达。嗜酸乳杆菌NCFM与Caco-2细胞共培养0、0.5、1、2和4 h，提取细胞蛋白质，采用Western Blot方法检测NF-κB的磷酸化水平；用NF-κB的特异性抑制剂PDTC预处理Caco-2细胞30 min，然后加入嗜酸乳杆菌NCF

Abstract:

Abstract：[Objective] To study the expression of immunity and inflammatory mediator factor PTX3 in intestinal epithelial cells treated with Lactobacillus acidophilus NCFM and further to reveal the regulatory mechanism. [Methods] Caco-2 cells were cocultured with Lactobacillus acidophilus NCFM for 0, 2, 4, 8, 12 h and 0, 0.5, 1, 2, 4 h respectively, then the total RNA and protein were extracted. The expression of PTX3 gene was analyzed by Real Time RT-PCR. The phosphorylation levels of NF-κB was analyzed by Western Blot. Caco-2 cells were pretreated with PDTC for 30 min before cocultured with Lactobacillus acidophilus NCFM for 2 h, then the total RNA was extracted and the expression of PTX3 gene was analyzed by Real Time RT-PCR. [Results] Lactobacillus acidophilus NCFM could induce the expression of PTX3 in Caco-2 cells. The PTX3 expression peaked at 4 h after coculture. Then its expression gradually waned out. Lactobacillus acidophilus NCFM could rapidly activate the phosphorylation of NF-κB, and the expression of PTX3 was decreased notably after pretreated with PDTC for 30 min. [Conclusion] Lactobacillus acidophilus NCFM could transiently regulate the immunity and inflammatory mediator factor PTX3 expression through rapidly activating NF-κB signaling pathway in Caco-2 cells.