食淀粉乳杆菌联合芦丁干预PoRV感染后肠道紧密连接蛋白及其相关因子变化

Changes of intestinal tight junction protein and its related factors afterLctobacillus amylovorus combined with rutin intervention after PoRV infection

通过益生菌联合多酚共同对抗猪轮状病毒(Porcine rotavirus,PoRV)感染,利用食淀粉乳杆菌和芦丁干预PoRV感染清洁级中国昆明鼠乳鼠,分为正常对照组(Control组)、病毒组(RV组)、食淀粉乳杆菌预防组(SR组)、芦丁1 mg·kg^(-1)预防组(LR组)、芦丁20 mg·kg^(-1)预防组(HR组)、食淀粉乳杆菌联合1 mg·kg^(-1)芦丁预防组(SLR组),应用实时荧光定量PCR和免疫组化检测Occludin和Claudin-4 mRNA相对表达量及蛋白分布,ELISA定量检测RV、胰蛋白酶、PAR-2、TGF-β和MMP-9含量。结果表明,RV组病毒含量最高,RV组Occludin mRNA相对表达量显著低于其他组,Claudin-4 mRNA相对表达量显著高于其他预防组,Claudin-4蛋白在肠道组织中分布不均;其他组Claudin-4 mRNA相对表达量均降低,但蛋白分布无改变。RV组胰蛋白酶、PAR-2、TGF-β和MMP-9含量均高于其他组;SLR组TGF-β含量低于SR组,MMP-9含量低于SR组。说明食淀粉乳杆菌和芦丁通过增加Occludin蛋白表达,抑制Claudin-4蛋白过表达及异常分布,下调胰蛋白酶、PAR-2、TGF-β及MMP-9含量,改善感染PoRV后肠道内相关因子异常分泌,减弱肠道损伤。

In this study,probiotics and polyphenols were used to jointly fight the infection of porcine rotavirus(PoRV),and Lactobacillus amylovorus and rutin were used to intervene the infection of PoRV in clean grade Kunming mice.The mice were divided into normal control group(Control group),virus group(RV group),Lactobacillus amylovorus prevention group(SR group),rutin 1 mg·kg^(-1) prevention group(LR group),rutin 20 mg·kg^(-1) prevention group(HR group)Lactobacillus amylovorus combined with 1 mg·kg^(-1) rutin prevention group(SLR group)was tested.The relative expression and protein distribution of Occludin and Claudin-4 mRNA were detected by real-time fluorescent quantitative PCR and immunohistochemistry,and the contents of RV,trypsin,PAR-2,TGF-βand MMP-9 were detected by ELISA.The results showed that the content of virus in RV group was the highest,the relative expression of Occludin mRNA in RV group was significantly lower than that in other groups,and the relative expression of Claudin-4 mRNA was significantly higher than that in other prevention groups.The distribution of Claudin-4 protein in intestinal tissue was uneven;other groups could reduce the relative expression of Claudin-4 mRNA,but the protein distribution did not change.The contents of trypsin,PAR-2,TGF-βand MMP-9 in RV group were higher than those in other groups;the contents of TGF-βand MMP-9 in SLR group was lower than that in SR group.It showed that Lactobacillus amylovorus and rutin inhibited the overexpression and abnormal distribution of Claudin-4 protein by increasing the expression of Occludin protein,down-regulate the contents of trypsin,PAR-2,TGF-βand MMP-9,and improve the abnormal secretion of related factors in the intestine after infection with PoRV to weaken the intestinal injury.