Lv Xuena, Man Chaoxin, Han Linlin, Wang Mingna, Zhang Guanghui, Liu Ying, Yang Shiqin, Xue Yuqing, Jiang Yujun
Key Lab of Dairy Science, Ministry of Education, College of Food Science and Engineering, Northeast Agricultural University, Harbin 150030, China.
Wei Sheng Wu Xue Bao. 2011 Apr;51(4):554-60.
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.
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-kappaB 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.
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-kappaB, and the expression of PTX3 was decreased notably after pretreated with PDTC for 30 min.
Lactobacillus acidophilus NCFM could transiently regulate the immunity and inflammatory mediator factor PTX3 expression through rapidly activating NF-kappaB signaling pathway in Caco-2 cells.
研究嗜酸乳杆菌NCFM处理的肠上皮细胞中免疫和炎症介质因子PTX3的表达，进一步揭示其调控机制。
将Caco-2细胞分别与嗜酸乳杆菌NCFM共培养0、2、4、8、12小时和0、0.5、1、2、4小时，然后提取总RNA和蛋白质。通过实时荧光定量逆转录聚合酶链反应（Real Time RT-PCR）分析PTX3基因的表达。通过蛋白质免疫印迹法（Western Blot）分析核因子κB（NF-κB）的磷酸化水平。在与嗜酸乳杆菌NCFM共培养2小时前，用PDTC预处理Caco-2细胞30分钟，然后提取总RNA，通过实时荧光定量逆转录聚合酶链反应分析PTX3基因的表达。
嗜酸乳杆菌NCFM可诱导Caco-2细胞中PTX3的表达。共培养后4小时PTX3表达达到峰值，然后其表达逐渐减弱。嗜酸乳杆菌NCFM可迅速激活NF-κB的磷酸化，用PDTC预处理30分钟后PTX3的表达显著降低。
嗜酸乳杆菌NCFM可通过快速激活Caco-2细胞中的NF-κB信号通路，瞬时调节免疫和炎症介质因子PTX3的表达。
