益生菌增加IPEC-J2细胞表面半乳糖在抑制RV感染中的作用

摘要

轮状病毒通过识别细胞表面糖链附着于肠上皮细胞,一些肠道内常驻菌群可以修改细胞表面糖链.以3种益生菌上清孵育猪小肠上皮细胞IPEC-J2,猪轮状病毒(PRV)感染细胞.采用凝集素荧光技术测定各处理组IPEC-J2细胞表面半乳糖,检测对应轮状病毒滴度和RV含量变化.结果表明,对照组PRV TCID50· 0.1 mL-1为10-7.15±0.14,干酪乳杆菌组TCID50·0.1mL-1为10-3.8755±0.125,食淀粉乳杆菌PRV TCID50·0.1 mL-1为10-4.125±0.138,枯草芽孢杆菌PRV TCID50· 0.1 mL-1为10-4.16±0.144.3组益生菌上清组在RV感染48 h内,RV量显著低于未处理组.凝集素荧光标记表明,3株益生菌上清均可改变细胞表面半乳糖含量.干酪乳杆菌、食淀粉乳杆菌和枯草芽孢杆菌上清可增加IPEC-J2细胞表面半乳糖含量,提高细胞抗RV感染能力,阻断RV黏附,保护肠道上皮细胞.%Rotaviruses attached to intestinal cells requires glycan recognition.Some bacteria from the gut microflora have been shown to modify cell-surface glycans.In this study,IPEC-J2 cultured cells were incubated with supernatants of probiotics and then infected with rotavirus.The porcine small intestinal epithelial cell line IPEC-J2 was incubated with three probiotic supernatants and then infected with porcine rotavirus (PRV).The surface galactose of IPEC-J2 cells was measured through lectin fluorescence technique,and the virulence of rotavirus was detected in each treatment group.The results showed that the PRV TCID50· 0.1 mL-1 of control group,Lactobacillus casei goup,Lactobacillus amylovorus group and Bacillus Subtilis group were 10-7.15 ± 0.14,10-3.875 ± 0.125,10-4.125 ± 0.138 and 10-4.16 ± 0.144.The RV content of the three probiotics group was significantly lower than that of the untreated group within 48 h of RV infection.Lectin fluorescence labeling indicated that the three probiotics supernatants could change the content of galactose on the surface of the cells.Lactobacillus casei supernatant,Lactobacillus amylovorus supernatant and Bacillus subtilis supernatant could increase the content of galactose on the surface of IPEC-J2 cells,improve the ability of anti-RV infection,inhibit the adhesion of the virus,and thus play a role in the protection of intestinal epithelial cells.