罗伊氏乳杆菌抑制肠上皮细胞炎症反应和凋亡的作用及机制

Inhibitory Effect and Mechanism of Lactobacillus reuteri on Inflammation and Apoptosis of Intestinal Epithelial Cells

目的:探讨罗伊氏乳杆菌对肠上皮细胞炎症反应和凋亡的作用及相关机制。方法:通过免疫荧光和酶联免疫吸附检测罗伊氏乳杆菌对肠上皮细胞炎症反应和凋亡的影响;采用生物信息学分析、定量聚合酶链式反应和双荧光素酶报告基因等实验筛选罗伊氏乳杆菌发挥作用的下游微小核糖核酸(microRNA,miRNA);通过双荧光素酶报告基因和Western blot等实验筛选miR-196a下游靶基因及其功能。结果:罗伊氏乳杆菌能够抑制脂多糖引起的肠上皮细胞炎症反应和凋亡,进一步分析发现罗伊氏乳杆菌通过上调肠上皮细胞miR-196a抑制细胞炎症和凋亡,miR-196a通过靶向抑制程序性细胞死亡因子4(programmed cell death 4,PDCD4)基因表达发挥对肠上皮细胞的保护作用。结论:罗伊氏乳杆菌通过miR-196a靶向抑制PDCD4基因表达,减缓肠上皮细胞炎症反应和凋亡。

Objective:To investigate the effect and mechanism of Lactobacillus reuteri on the inflammatory response and apoptosis of intestinal epithelial cells.Methods:Immunofluorescence and enzyme linked immunosorbent assay(ELISA)were used to detect the effect of L.reuteri on the inflammatory response and apoptosis of intestinal epithelial cells.Bioinformatics analysis,real-time polymerase chain reaction(PCR)and the dual-luciferase reporter gene assay were used to analyze downstream miRNA expression under the action of L.reuteri.The downstream target gene of miR-196a and its function were investigated using the dual-luciferase reporter gene assay and Western blot(WB).Results:L.reuteri inhibited inflammatory responses and apoptosis induced by lipopolysaccharide(LPS)in intestinal epithelial cells by up-regulating miR-196a.miR-196a could protect intestinal epithelial cells by targeting the expression of programmed cell death factor 4(PDCD4).Conclusion:L.reuteri could inhibit the expression of the PDCD4 gene through miR-196a,thereby alleviating the inflammatory response and apoptosis of intestinal epithelial cells.