苏氨酸对体外培养感染伪狂犬病毒猪空肠上皮细胞免疫相关基因表达的影响

本研究旨在探讨苏氨酸对体外培养感染伪狂犬病毒(PRV)猪空肠上皮细胞免疫相关基因表达的影响.试验选用IPEC-J2细胞为细胞模型,分为4个处理,对照组为未经PRV感染、苏氨酸浓度53.45 mg/L,Ⅰ、Ⅱ、Ⅲ组为试验组,均经PRV感染,苏氨酸浓度分别为53.45、106.90、160.35 mg/L,每组3个重复,每个重复4孔.37℃、5％ CO2培养后1、6、12、24 h每组取1个重复采集细胞,用实时荧光定量PCR方法检测白介素1β(IL-1β)、白介素6(IL-6)、白介素15(IL-15)、肿瘤坏死因子α(TNF-α)及转化生长因子β1( TGF-β1) mRNA表达量.结果表明:1)PRV感染极显著降低了IL-1β mRNA表达量(P =0.001),苏氨酸水平(P=0.830)、时间与苏氨酸水平交互作用(P =0.249)的影响均不显著；2)PRV感染对IL-6mRNA表达量影响不显著(P =0.162),随苏氨酸水平升高,IL-6 mRNA表达量极显著地先升高后降低(P=0.002),时间与苏氨酸水平交互作用的影响显著(P =0.012)；3)PRV感染极显著提高了IL-15 mRNA表达量(P =0.000),提高苏氨酸水平极显著提高了IL-15 mRNA表达量(P=0.000),时间与苏氨酸水平交互作用的影响亦极显著(P =0.000)；4) PRV感染极显著提高了TNF-α mRNA表达量(P=0.000),苏氨酸水平(P =0.113)、时间与苏氨酸水平交互作用(P =0.195)的影响均不显著；5)PRV感染极显著提高了TGF-β1 mRNA表达量(P=0.000),时间与苏氨酸水平交互作用的影响显著(P =0.015),苏氨酸水平的影响不显著(P=0.055).结果提示,补充苏氨酸对感染PRV的IPEC-J2细胞先天性免疫功能具有分子表达水平的调控作用,总体上能够抑制IL-1β、TNF-α、TGF-β1基因表达,加强IL-6、IL-15基因表达,但影响具有时间效应.

Effects of Threonine on Expression of Immune-Related Genes of Porcine Intestinal Epithelial Cell Infected with Pseudorabies Virus in Vitro

The present study was conducted to evaluate the effects of threonine(Thr) on expression of immune-related genes of porcine intestinal epithelial cell infected with pseudorabies virus(PRV) in vitro.IPEC-J2 cells were selected as the cell model,and were divided into four treatments.Cells in the control group were un-infected with PRV and cultured with Thr at the concentration of 53.45 mg/L in culture medium,and cells in experimental groups(groups Ⅰ,Ⅱ and Ⅲ) were infected with PRV and cultured with Thr at concentrations of 53.45,106.90 and 160.35 mg/L in culture medium,respectively.There were 3 replicates in each group and 4 wells per replicate.After cultured in 37 ℃ and with 5% CO2 for 1,6,12 and 24 h,cells in one replicate from each group were collected,respectively.Then mRNA expression levels of interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-15(IL-15),tumor necrosis factor-α(TNF-α) and transforming growth factor-β1(TGF-β1) in those cells were determined by real-time PCR.The results showed as follows: 1) IL-1β mRNA expression level was significantly reduced by PRV infection(P=0.001),however effects of Thr level(P=0.830) and its interaction with time(P=0.249) were not significant;2) IL-6 mRNA expression level was not significantly reduced by PRV infection(P=0.162),while was significantly increased at first and then decreased with the increasing of Thr level(P=0.002),and the interaction between time and Thr level was significant(P=0.012);3) PRV infection significantly increased the mRNA expression of IL-15(P=0.000),and the expression level was significantly increased with the increasing of Thr level(P=0.000),the interaction between time and Thr level was also significant(P=0.000);4) PRV infection significantly increased the mRNA expression level of TNF-α(P=0.000),however effects of Thr level(P=0.113) and its interaction with time(P=0.195) were not significant;5) PRV infection significantly increased the mRNA expression level of TGF-β1(P=0.000),and the interaction between Thr level and time was significant(P=0.015),but the effect of Thr level was not significant(P=0.055).The results indicate that Thr supplementation can regulate the innate immunity function of IPEC-J2 cells infected with PRV at molecular level,and can generally inhibit gene expression of IL-1β,TNF-α and TGF-β1,but enhance gene expression of IL-6 and IL-15,while the effects vary with the time.