HSP27 基因表达对仔猪抗大肠杆菌感染的影响。

Key Laboratory for Animal Genetics, Breeding, Reproduction and Molecular Design of Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, PR China.

Key Laboratory for Animal Genetics, Breeding, Reproduction and Molecular Design of Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, PR China; Joint International Research Laboratory of Agriculture & Agri-Product Safety, Yangzhou University, Yangzhou, Jiangsu, PR China.

Gene. 2021 Mar 20;773:145415. doi: 10.1016/j.gene.2021.145415. Epub 2021 Jan 12.

Heat shock protein 27 (HSP27) plays an important role in protecting cells from various stress factors. This study aimed to investigate the function of HSP27 gene and its regulatory mechanism as infected by Escherichia coli (E. coli) at the tissue and cellular levels. Real-time PCR was used to detect the differential expression of HSP27 gene in F18 resistant and sensitive Sutai pigs and the differential expression upon E. coli F18ab, F18ac, K88ac bacterial supernatant, thallus infection and LPS induction in IPEC-J2. In addition, the HSP27 gene overexpression vector was constructed to detect the effect of the HSP27 gene overexpression on the adhesion of E. coli F18 to IPEC-J2, secretion of pro-inflammatory factors, and the expression of the upstream key genes in Mitogen-activated protein kinase (MAPK) pathway. Ribosomal S6 kinase (RSK2) is an important protein in the MAPK pathway. Therefore, the RSK2 gene overexpression vector was constructed and the number of colonies was counted after co-transfection of HSP27 and RSK2 gene. Results revealed that the expression level of HSP27 gene in resistant individuals in 11 tissues was higher than sensitive type. At the cellular level, the relative expression levels of HSP27 gene were increased after F18ab, F18ac bacterial supernatant, F18ab thallus infection, and LPS induction for 4 h (P < 0.01). The adhesion ability of E. coli F18ab to IPEC-J2 was significantly reduced after HSP27 gene overexpression (P < 0.01), and the concentration of pro-inflammatory factors in the HSP27 gene overexpression group was significantly reduced compared with the control group after F18ab infection (P < 0.05). Furthermore, the expression of RSK2 was significantly increased in HSP27 overexpression group upon F18ab infection (P < 0.01). The colonies quantitative results also showed that the number of colonies was significantly reduced after co-transfection of HSP27 and RSK2 gene. We indicated that the high expression of HSP27 gene may resist the inflammatory response caused by exogenous stress and enhance the ability of IPEC-J2 to resist E. coli F18 infection. RSK2 gene in the MAPK pathway may cooperate with HSP27 gene to participate in the immune response of the organism, which provides a theoretical basis for the study of the mechanism of anti-E. coli infection in piglets.

热休克蛋白 27（HSP27）在保护细胞免受各种应激因素方面发挥着重要作用。本研究旨在从组织和细胞水平上探讨 HSP27 基因的功能及其调控机制，当感染大肠杆菌（E. coli）时。实时 PCR 用于检测 F18 抗性和敏感 Sutai 猪中 HSP27 基因的差异表达，以及 E. coli F18ab、F18ac、K88ac 细菌上清液、菌毛感染和 LPS 诱导后 IPEC-J2 中的差异表达。此外，构建 HSP27 基因过表达载体，检测 HSP27 基因过表达对 F18 大肠杆菌与 IPEC-J2 黏附、促炎因子分泌以及丝裂原活化蛋白激酶（MAPK）通路上游关键基因表达的影响。核糖体 S6 激酶（RSK2）是 MAPK 通路中的重要蛋白。因此，构建 RSK2 基因过表达载体，共转染 HSP27 和 RSK2 基因后计数菌落数。结果表明，11 种组织中抗性个体 HSP27 基因的表达水平高于敏感型。在细胞水平上，F18ab、F18ac 细菌上清液、F18ab 菌毛感染和 LPS 诱导 4 h 后 HSP27 基因的相对表达水平均升高（P<0.01）。HSP27 基因过表达后，E. coli F18ab 对 IPEC-J2 的黏附能力显著降低（P<0.01），且 F18ab 感染后 HSP27 基因过表达组促炎因子浓度较对照组显著降低（P<0.05）。此外，F18ab 感染后 HSP27 过表达组 RSK2 的表达显著增加（P<0.01）。菌落定量结果也表明，共转染 HSP27 和 RSK2 基因后，菌落数明显减少。我们表明 HSP27 基因的高表达可能抵抗外源性应激引起的炎症反应，增强 IPEC-J2 抵抗大肠杆菌 F18 感染的能力。MAPK 通路中的 RSK2 基因可能与 HSP27 基因协同参与机体的免疫反应，为仔猪抗大肠杆菌感染机制的研究提供了理论依据。