猪瘟病毒对ST细胞NF-κB信号通路炎性因子表达量的影响

Effect of CSFV Infection on mRNA Expressions of NF-κB Signaling Pathway Related Factors in ST Cells

探讨猪瘟病毒(CSFV)感染猪睾丸上皮细胞系(ST)对核因子-κB(NF-κB)信号通路相关炎性因子的表达。分别收集CSFV感染4、8、12、16、24、48 h的ST细胞,建立正常对照组和CSFV感染各试验组。荧光定量PCR(qPCR)法检测NF-кB通路相关炎性因子CHUK、IKBKB、NFKBIA、RelA基因mRNA表达量的变化,Western blot检测p65蛋白在核中的表达。NFKBIA、IKBKB基因的转录水平差异显著(P<0.05);RelA基因的表达差异不显著(P>0.05);CHUK基因在ST细胞感染CSFV后,转录水平差异显著(P<0.05),Western blot试验显示,p65入核。CSFV感染ST细胞后,NF-κB的经典信号通路和非经典信号通路的均被激活,初步探索了机体感染CSFV对NF-κB通路上、下游的影响,为猪瘟的有效防控提供理论依据。

In order to explore the classical swine fever virus(CSFV) infection of porcine testicular epithelial cells(ST) on nuclear factor-kappa B(NF-κB) signaling pathway, an in vitro infection as a model was established, ST cells infected with CSFV at 4,8,12,16,24,48 hours were collected to establish a normal control group and CSFV infected experimental groups.Fluorescence quantitative PCR(qPCR) method was used to detect the changes of mRNA expressions of NF-кB pathway-related inflammatory genes CHUK,IKBKB,NFKBIA and RelA.Western blot was used to detect the expression of p65 protein in the nucleus.The results showed that the transcriptional levels of NFKBIA and IKBKB genes were significantly higher(P<0.05),and the expression of RelA gene did not change significantly(P>0.05).After CSFV infection of ST,the mRNA expression of CHUK gene was significantly higher(P<0.05),both the classical and non-classical signaling pathways of NF-κB were activated.The study preliminarily explored the mechanism by which classic swine fever virus affects the body's NF-κB signaling pathway, and provided a theoretical basis for effective prevention and control of classic swine fever.