食淀粉乳杆菌对猪A群轮状病毒NSP4在小鼠肠组织表达的影响

Effects of Lactobacillus amylovorus on the expression of non-structural protein NSP4 of porcine group A rotavirus in intestinal tissue of mice

为探究食淀粉乳杆菌对猪轮状病毒(PRV)非结构蛋白NSP4表达的影响,对3日龄清洁级BALB/c小鼠分别在感染PRV前后灌服食淀粉乳杆菌,应用实时荧光定量PCR检测感染病毒后空肠组织中PRVNSP4的mRNA相对表达量,采用比色法测定空肠组织内钙离子含量变化,并通过ELISA定量检测空肠组织中整联蛋白α2β1、磷脂酶C和Dicer的含量。结果显示,饲喂食淀粉乳杆菌第5天和第7天时,预防组和辅疗组中NSP4的mRNA相对表达量显著低于病毒组(P<0.05);除预防组第11天外,感染病毒后各检测时间点预防组和辅疗组的钙离子含量显著低于病毒组(P<0.05);饲喂食淀粉乳杆菌第7天、第9天和第11天时,预防组和辅疗组中整联蛋白α2β1的含量显著低于病毒组(P<0.05);感染病毒后各检测时间点预防组和辅疗组中磷脂酶C和Dicer的含量均低于病毒组。以上结果表明:在小鼠感染轮状病毒前后灌服食淀粉乳杆菌均能显著抑制PRVNSP4的mRNA表达和整联蛋白α2β1的合成,并降低空肠组织中钙离子,磷脂酶C和Dicer的含量,这为研究益生菌抗轮状病毒机制提供参考。

To investigate the effect of Lactobacillus amylovorus on the mRNA expression of non-structural protein NSP4 in mice infected by porcine rotavirus(PRV),three-day-old clean grade BALB/c mice were fed with L.amylovorus before or after PRV infection.The relative mRNA expression levels of NSP4 in the jejunal tissues of mice infected with PRV were detected by real-time fluorescence quantitative PCR.The calcium ion content in the jejunal tissues were determined by colorimetry,and the content of integrinα2β1,phospholipase C and Dicer in the jejunal tissues were quantitatively detected by ELISA.The results showed that the relative mRNA expression levels of NSP4 in prevention and auxiliary treatment groups were significantly lower than that in rotavirus group on the fifth and seventh day after exposed to L.amylovorus(P<0.05).Except for the prevention group on the eleventh day,the calcium ion content in the prevention and auxiliary treatment groups were significantly lower than that in the rotavirus group at each detection time points with infected PRV(P<0.05).All of the content of integrinα2β1 in prevention and auxiliary treatment groups at seven,nine and eleven days post-exposed to L.amylovorus were significantly lower than that in the rotavirus group(P<0.05).The content of phospholipase C and Dicer in prevention and auxiliary treatment groups at each detection time point with infected PRV were lower than that in the rotavirus group.The results indicated that L.amylovorus can significantly inhibit the mRNA expression of PRV NSP4 and the synthesis of integrinα2β1,and can reduce the content of calcium ion,phospholipase C and Dicer,which will provide reference for studying the mechanism of probiotics against rotavirus.