摘要
目的观察甘露消毒丹及其挥发油对流感病毒感染小鼠血清中白细胞介素4(IL-4)、肿瘤坏死因子α(TNF-α)、干扰素α/β(IFN-α/β)含量及肺组织中非结构蛋白1(NS1)、维甲酸诱导基因Ⅰ(RIG-Ⅰ)mRNA、核因子κB(NF-κB)蛋白的影响,以探讨甘露消毒丹及其挥发油抗病毒作用的分子机制。方法选择SPF级BALB/c小鼠36只,雌雄各半,随机分为6组,分别为正常组、模型组、灌胃组、滴鼻组、灌胃滴鼻合用组、奥司他韦组,每组6只。除正常组外,其余5组以流感病毒FM1-6-E2滴鼻造模。于造模后第7天,摘眼球放血处死小鼠,通过透射电子显微镜对肺组织进行检测以及图像分析,采用ELISA法测定血清中IL-4、TNF-α、IFN-α/β含量,qRT-PCR检测各组小鼠肺组织中NS1、RIG-ⅠmRNA的表达,Western blot方法测定各组小鼠肺组织NF-κB蛋白的表达。结果 (1)流感病毒感染后模型小鼠肺组织超微结构可见Ⅱ型细胞坏死、肺泡结构破坏,肺泡腔内纤毛增粗、脱落,形成肺部炎症;各治疗组均可改善肺泡Ⅱ型上皮细胞超微结构的异常改变。(2)流感病毒感染小鼠后,血清中IL-4、TNF-α、IFN-α/β的含量及肺组织中的NS1、RIG-ⅠmRNA、NF-κB蛋白表达均高于正常组,模型组与正常组比较,差异显著;与模型组比较,灌胃组、滴鼻组、灌胃滴鼻合用组、奥司他韦组血清中IFN-α/β含量显著升高,IL-4、TNF-α含量及NS1 mRNA、RIG-ⅠmRNA、NF-κB蛋白表达明显下调(P<0.05),甘露消毒丹各治疗组不同剂型抗流感病毒作用差异有统计学意义(P<0.05),灌胃滴鼻合用组>灌胃组>滴鼻组。结论甘露消毒丹具有抗流感病毒感染的作用,通过抑制RIG-Ⅰ/NF-κB信号传导通路的活化,阻止炎症的病理反应,提高免疫因子含量,降低流感病毒引起的免疫炎性损伤。
To observe the effects of Ganlun Xiaodu micropill(GXM)and its volatile oil on the levels of IL-4,TNF-α,IFN-α/βand NS1mRNA,RIG-ⅠmRNA and NF-κB protein in lung tissue of mice infected with influenza virus(IV),in order to explore the molecular mechanism of the antiviral effect of GXM and its volatile oil.MethodsA total of36SPF BALB/c mice,half female and half male,were randomly divided into6groups,with6rats in each group.They were normal group(A group),model group(B group),gavage group(C group),intranasal group(D group),combination group(E group)and oseltamivir group(F group).Except the normal group,the remained5groups were modeled with influenza virus FM16E2.On the seventh day after the model,the mice were killed by picking up the eyeball and releasing blood.The lung tissue was detected by transmission electron microscope and the image was analyzed.The content of IL-4,TNF-αand IFN-α/βin serum was determined by ELISA,and the expression of NS1and RIG-ⅠmRNA in lung tissue of mice was detected by qRT-PCR.Western blot method was used to determine the expression of NF-κB protein in lung tissues of mice.Results(1)The ultrastructure of lung tissue in the model mice after IV infection showed necrosis of typeⅡcells,destruction of alveolar structure,thickening and shedding of cilium in the alveoli,and pulmonary inflammation was formed.All the treatment groups were improved in the ultrastructural abnormal change of the alveolar typeⅡepithelial cells.(2)After IV infection in mice,the content of IL-4,TNF-αand IFN-α/βin serum and the expression of NS1mRNA,RIG-ⅠmRNA and NF-κB protein in lung tissues were all higher than those in A group.The difference was significant between B group and A group;Compared with the B group,the serum levels of IFN-α/βin the serum of C,D,E and F groups increased significantly,while the content of IL-4,TNF-αand the expression of NS1mRNA,RIG-ⅠmRNA and NF-κB protein were obviously down regulated(P<0.05).There was significant difference in the anti IV effect among different dosage forms of GXM treatment groups(P<0.05),the effect being E group>C group>D group.ConclusionGXM has the function of anti IV infection.It inhibits the pathological reaction of inflammation,enhances the content of immune factors and reduces the immune inflammatory injury induced by IV by inhibiting the activation of RIG-Ⅰ/NF-κB signaling pathway.
作者
刘光华
刘娟
高畅
张聪聪
贾晓妍
LIU Guanghua;LIU Juan;GAO Chang;ZHANG Congcong;JIA Xiaoyan(Medical Department of Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China)
出处
《中国中西医结合儿科学》
2017年第6期461-466,共6页
Chinese Pediatrics of Integrated Traditional and Western Medicine
基金
国家自然基金青年项目(81403293)
