摘要
为了比较硝唑尼特在调控日本脑炎病毒(JEV)和脂多糖(LPS)诱导的炎症反应上的异同,分别用感染复数(MOI)=10的JEV和1μg/mL的LPS体外诱导RAW264.7细胞构建炎症模型并给予不同浓度的硝唑尼特处理。采用实时荧光定量PCR、Western blot和ELISA等方法检测硝唑尼特对病毒复制、多种炎性细胞因子以及c-Jun氨基末端激酶(JNK)通路蛋白的调控情况。结果显示:JEV和LPS诱导后都能显著上调细胞内炎症因子的表达(P<0.01);给予硝唑尼特处理的细胞JEV复制减少,炎症因子如肿瘤坏死因子α(TNF-α)、白介素1β(IL-1β)的mRNA表达下降,但不能有效抑制JEV引起的细胞内诱导型一氧化氮合成酶(iNOS)和环氧合酶2(COX-2)的表达,同时也不能下调磷酸化JNK蛋白(pJNK)的表达;而在LPS诱导的模型中,硝唑尼特对LPS引起的TNF-α、iNOS等炎症因子表达量升高具有显著的抑制作用(P<0.05),且能下调JNK蛋白的磷酸化水平(P<0.05)。以上结果显示了硝唑尼特对JEV和LPS诱导的炎症反应都具有一定的抑制作用,但其调控炎症的方式存在较大差异,这为硝唑尼特通过调节天然免疫而发挥抗多种病原体的作用机制研究提供数据支持。
To compare the modulation of nitazoxanide on inflammatory responses induced by Japanese encephalitis virus(JEV)and lipopo-lysaccharide(LPS),mice were used and treated with JEV(MOI=10)and LPS(1μg/mL)to induce an inflammation model in RAW2647 cells and were given different concentrations of nitazoxanide for treatment.Then,real time PCR,Western blot and ELISA were used to determine the change of viral replication,multiple inflammatory cytokines as well as the expression level of JNK pathway proteins.The results showed that both JEV and LPS significantly up-regulated the expression of intracellular inflammatory cytokines(P<0.01).The cells treated with nitazoxanide were found to have reduced viral replication and the expression levels of TNF-αand IL-1βmRNA.However,the results also showed that nitazoxanide could not effectively inhibit the expression levels of intracellular iNOS and COX-2 caused by JEV and could not downregulate the phosphorylation of JNK protein,either.Yet,in the LPS induced model,nitazoxanide was effective against TNF-αand iNOS(P<0.05),and it downregulated(P<0.05)the phosphorylation of the JNK protein.The above results indicated that nita-zoxanide could inhibit inflammatory responses induced by both JEV and LPS,but there were evident differences in the ways it regulated in-flammation,which would serve as support for studying how nitazoxanide exerts its effect against multiple pathogens by modulating innate im-munity.
作者
苏钰
陈栋梁
马慧
王霄旸
王春梅
周文
张可煜
SU Yu;CHEN Dongliang;MA Hui;WANG Xiaoyang;WANG Chunmei;ZHOU Wen;ZHANG Keyu(Shanghai Veterinary Research Institute,CAAS Key Laboratory of Veterinary Chemical Drugs and Pharmaceutics,Ministry of Agriculture and Rural Affairs,Shanghai 201100,China;Yunnan Agricultural University,Kunming 650000,China)
出处
《畜牧与兽医》
CAS
北大核心
2023年第10期21-28,共8页
Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金项目(31872516,32172913)
国家重点研发计划项目(2018YFE0192600)。
关键词
硝唑尼特
日本脑炎病毒
脂多糖
炎症因子
nitazoxanide
Japanese encephalitis virus
lipopolysaccharide
inflammation cytokines
