经口暴露于单端孢霉烯族呕吐毒素(脱氧雪腐镰刀菌烯醇)后小鼠体内细胞因子mRNA表达的差异:剂量反应和时间进程

Differential cytokine mRNA expression in mice after oral exposure to the trichothecene vomitoxin (deoxynivalenol): dose response and time course.

作者信息

Zhou H R, Yan D, Pestka J J

机构信息

Department of Food Science and Human Nutrition, Michigan State University, East Lansing 48824, USA.

出版信息

Toxicol Appl Pharmacol. 1997 Jun;144(2):294-305. doi: 10.1006/taap.1997.8132.

Abstract

Acute oral exposure of B6C3F1 mice to vomitoxin (VT) has been previously shown to induce expression of mRNAs for cytokines that are characteristically produced in lymphoid tissues by macrophage and T cells. The purpose of this study was to evaluate the effects of VT dose on the expression of mRNAs for a cytokine profile consisting of TNF-alpha, IL-1beta, IL-6, IL-12, IFN-gamma, IL-2, IL-4, IL-5, IL-10, IL-12, and TGF-beta and to measure the kinetics of these responses. The effects of a single oral exposure to 0, 0.1, 0.5, 1, 5, and 25 mg/kg BW of VT on cytokine mRNA abundance in spleen and Peyer's patches (indicators of systemic and mucosal immune compartments, respectively) were assessed at 2 hr postexposure using reverse transcriptase-polymerase chain reaction (RT-PCR) in combination with hybridization analysis. Both 5 and 25 mg/kg VT significantly induced the mRNAs for the proinflammatory cytokines IL-1beta, IL-6, and TNF-alpha; the T helper 1 cytokines IFN-gamma and IL-2; and the T helper 2 cytokines IL-4 and IL-10, whereas lower doses had no effect. IL-12p40 mRNA was also induced but not IL-12p35 mRNA. The effects were more pronounced in spleen than in the Peyer's patches. IL-5 and TGF-beta mRNAs were expressed constitutively in spleen and Peyer's patches but were not affected by VT. When cytokine mRNA levels were measured at 1, 2, 4, 8, and 24 hr after oral exposure to 25 mg/kg BW of VT, mRNA expression kinetics varied among cytokines or between spleen and Peyer's patches. The duration of elevated mRNA expression in spleen was 1-8 hr for TNF-alpha, IL-6, IFN-gamma, and IL-10 and was 1-4 hr for IL-1beta, IL-12p40, IL-2, and IL-4. In Peyer's patches, duration periods were 1-8 hr for IL-6, IL-2, and IL-10; 1-4 hr for IL-1beta, IL-12p40, and TNF-alpha; and 1-2 hr for IFN-gamma. Serum levels of TNF-alpha, IL-6, and IFN-gamma were elevated 3 hr after exposure to 25 mg/kg VT, thus suggesting that elevation of splenic and Peyer's patch mRNA abundance correlated with increases in systemic production of these cytokines. Taken together, the results indicate that a single VT exposure rapidly induces gene expression in vivo for a wide range of cytokines with apparently complete recovery occurring after 24 hr. Elevated cytokine expression may play an important role in the pathophysiologic effects of VT and other trichothecenes.

摘要

先前已表明,B6C3F1小鼠经口急性暴露于呕吐毒素(VT)可诱导巨噬细胞和T细胞在淋巴组织中典型产生的细胞因子的mRNA表达。本研究的目的是评估VT剂量对由肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、白细胞介素-12(IL-12)、干扰素-γ(IFN-γ)、白细胞介素-2(IL-2)、白细胞介素-4(IL-4)、白细胞介素-5(IL-5)、白细胞介素-10(IL-10)、IL-12和转化生长因子-β(TGF-β)组成的细胞因子谱mRNA表达的影响,并测量这些反应的动力学。在暴露后2小时,使用逆转录聚合酶链反应(RT-PCR)结合杂交分析,评估单次经口暴露于0、0.1、0.5、1、5和25mg/kg体重的VT对脾脏和派尔集合淋巴结(分别为全身和黏膜免疫区室的指标)中细胞因子mRNA丰度的影响。5和25mg/kg的VT均显著诱导促炎细胞因子IL-1β、IL-6和TNF-α的mRNA表达;辅助性T细胞1细胞因子IFN-γ和IL-2的mRNA表达;以及辅助性T细胞2细胞因子IL-4和IL-10的mRNA表达,而较低剂量则无影响。IL-12p40 mRNA也被诱导,但IL-12p35 mRNA未被诱导。这些影响在脾脏中比在派尔集合淋巴结中更明显。IL-5和TGF-β mRNA在脾脏和派尔集合淋巴结中组成性表达,但不受VT影响。当在经口暴露于25mg/kg体重的VT后1、2、4、8和24小时测量细胞因子mRNA水平时,mRNA表达动力学在不同细胞因子之间或在脾脏和派尔集合淋巴结之间有所不同。脾脏中mRNA表达升高的持续时间,TNF-α、IL-6、IFN-γ和IL-10为1至8小时,IL-1β、IL-12p40、IL-2和IL-4为1至4小时。在派尔集合淋巴结中,IL-6、IL-2和IL-10的持续时间为1至8小时;IL-1β、IL-12p40和TNF-α为1至4小时;IFN-γ为1至2小时。暴露于25mg/kg的VT后3小时,血清中TNF-α、IL-6和IFN-γ水平升高,因此表明脾脏和派尔集合淋巴结mRNA丰度的升高与这些细胞因子全身产生的增加相关。综上所述,结果表明单次VT暴露可在体内迅速诱导多种细胞因子的基因表达,24小时后显然完全恢复。细胞因子表达升高可能在VT和其他单端孢霉烯的病理生理效应中起重要作用。

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