Pathogenic E-coli HPI upregulate the expression of inflammatory factors in porcine small intestinal epithelial cells by ubiquitin proteasome pathway
Pathogenic E-coli HPI upregulate the expression of inflammatory factors in porcine small intestinal epithelial cells by ubiquitin proteasome pathway
复制标题
致病性大肠杆菌HPI通过泛素蛋白酶体途径上调猪小肠上皮细胞炎症因子的表达
DOI：
10.1016/j.rvsc.2018.08.009
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发表时间：
2018-10-01
影响因子：
2.4
通讯作者：
Gao, Hong
中科院分区：
文献类型：
--
作者：
Liu, Chaoying;Shan, Chunlan;Gao, Hong
目的探讨致病性大肠埃希菌高致病性岛(HPI)通过泛素蛋白酶体途径对炎性因子表达的影响。首先，成功构建了UBC-SUS-263 shRNA载体，并通过脂质体将其导入猪小肠上皮细胞(IPEC-J2)，沉默泛素基因。分别用HPI+和HPI-菌株感染IPEC-J2细胞。最后检测UBC-SUS-263 shRNA(Ub-shRNA)对IPEC-J2细胞内核因子-kappaB和IkB-α的表达及NE-kappa B、TNF-α和IL-1蛋白水平的影响。结果表明，Ub-shRNA能有效抑制IPEC-J2细胞的泛素化途径。HPI感染IPEC-J2细胞后，与对照组和HPI感染组相比，Ub-hsRNA转染的IPEC-J2细胞中NF-kappa B和I-kappa B-α的mRNA和蛋白水平均显著降低。与HPI感染组相比，HPI+感染组的下游细胞因子如TNF-α和IL-1的表达一直较高。但在正常的IPEC-J2细胞中，无论是HPI还是HPI-，都能诱导核因子-kappa B和I-kappaB-α及其下游细胞因子的表达增加。因此，大肠杆菌HPI可以上调IKB-α的表达，通过泛素化途径促进肿瘤坏死因子-α和IL-1的释放。
To investigate the effects of pathogenic Escherichia coli high pathogenicity island (HPI) on the expression of inflammatory factors via ubiquitin proteasome pathway. Firstly, the UBC-sus-263 shRNA plasmid was successfully established and transfected into porcine small intestine epithelial cells (IPEC-J2) by liposome to silence the ubiquitinntion gene. Then the IPEC-J2 was infected with E. coli HPI+ and HPI- strains, respectively. Finally, the mRNA of intracellular NF-kappa B and IKB-alpha, and the protein levels of NE-kappa B, TNF-alpha and IL-1 in IPEC-J2 cell line transfected with UBC-sus-263 shRNA (Ub-shRNA) were detected. The results showed that the Ub-shRNA was effectively inhibited ubiquitination pathway in the IPEC-J2 cell. After infected with HPI, the mRNA and protein levels of NF-kappa B and I kappa B-alpha were dramatically decreased in Ub-hsRNA transfected IPEC-J2 cells compared to the control and HPI--infected groups. Consistently, the production of downstream cytokines such as TNF-alpha and IL-1 were highly expressed after HPI+-infection than that of HPI--infected groups. However, whether the HPI or HPI-, both could induce increasingly expression of NF-kappa B and I kappa B-alpha and its downstream cytokines in normal IPEC-J2 cells. Thus, the E. coil HPI can upregulate the expression of IKB-alpha to promote the releasing of TNF-alpha and IL-1 via the ubiquitination pathway.