海藻多糖对仔猪小肠上皮细胞紧密连接蛋白及核转录因子-κB信号通路相关基因表达的影响

Effects of Seaweed Polysaccharides on Expression of Tight Junction Protein and Related Genes in Nuclear Transcription Factor⁃κB Signaling Pathway in Porcine Epithelial Cells

本试验旨在研究不同浓度海藻多糖对仔猪小肠上皮细胞(IPEC-J2细胞)紧密连接蛋白及核转录因子-κB(NF-κB)信号通路相关基因表达的影响。IPEC-J2细胞加入浓度分别为0(对照)、60、120和240μg/mL的海藻多糖处理培养基培养24 h,分析海藻多糖对IPEC-J2细胞机械屏障和免疫屏障相关基因以及蛋白表达的影响。结果表明:1)与对照组相比,60、120μg/mL的海藻多糖可以显著上调紧密连接蛋白闭锁蛋白(occludin)和闭合小环蛋白-1(ZO-1)的mRNA相对表达量(P<0.05);但240μg/mL海藻多糖组occludin和ZO-1的mRNA相对表达量与对照组无显著差异(P>0.05)。与对照组相比,120和240μg/mL的海藻多糖可以显著上调封闭蛋白-1(claudin-1)的mRNA相对表达量(P<0.05)。与对照组相比,120、240μg/mL的海藻多糖可以显著上调claudin-1和ZO-1的蛋白相对表达量(P<0.05)。2)与对照组相比,60、120和240μg/mL的海藻多糖均显著降低了髓样分化蛋白88(MyD88)和p65的mRNA相对表达量(P<0.05),240μg/mL的海藻多糖显著降低了Toll样受体-4(TLR-4)和核转录因子-κB抑制蛋白α(IκBα)的mRNA相对表达量(P<0.05)。由此可知,海藻多糖可以上调IPEC-J2细胞紧密连接蛋白occludin、claudin-1和ZO-1的mRNA相对表达量,降低IPEC-J2细胞NF-κB信号通路相关基因的表达,对改善仔猪小肠上皮细胞屏障功能具有积极的作用。

This study aimed to evaluate the effects of different concentration of seaweed polysaccharides on ex⁃pression of tight junction protein and related genes in nuclear transcription factor⁃κB(NF⁃κB)signaling path⁃way in porcine epithelial cells(IPEC⁃J2 cells).The IPEC⁃J2 cells were treated with seaweed polysaccharides culture medium with concentrations of 0(control),60,120 and 240μg/mL,cultured for 24 h,and the effects of seaweed polysaccharides on expression of mechanical barrier and immune barrier related gene and protein in IPEC⁃J2 cells were analysed.The results showed as follows:1)compared with the control group,60 and 120μg/mL seaweed polysaccharides significantly upregulated the mRNA relative expression levels of occludin and zonula occluden⁃1(ZO⁃1)(P<0.05);but no significant differences were observed in the mRNA relative expression levels of occludin and ZO⁃1 between control group and 240μg/mL seaweed polysaccharides group(P>0.05).Compared with control group,120 and 240μg/mL seaweed polysaccharides significantly upregulated the mRNA relative expression level of claudin⁃1(P<0.05).Compared with control group,120 and 240μg/mL seaweed polysaccharides significantly upregulated the protein relative expression levels of clau⁃din⁃1 and ZO⁃1(P<0.05).2)Compared with control group,60,120 and 240μg/mL seaweed polysaccha⁃rides significantly decreased the mRNA relative expression levels of myeloid differentiation protein 88(MyD88)and p65(P<0.05),and 240μg/mL seaweed polysaccharides significantly decreased the mRNA relative expression levels of Toll like receptor⁃4(TLR⁃4)and nuclear transcription factor inhibitor proteinα(IκBα)(P<0.05).In conclusion,seaweed polysaccharides can upregulate the mRNA relative expression lev⁃els of occludin,claudin⁃1 and ZO⁃1 in IPEC⁃J2 cells,and decrease the expression of NF⁃κB signaling pathway related gene in IPEC⁃J2 cells,and have positive effects on improv the barrier function of porcine epithelial cells.