表没食子儿茶素没食子酸酯对H9N2猪流感病毒诱导小鼠肺损伤中Toll样受体-4表达的影响

doi:10.11843/j.issn.0366-6964.2015.08.021

摘要/Abstract

摘要：

旨在探讨表没食子儿茶素没食子酸酯（EGCG）对猪源H9N2流感病毒感染诱导小鼠肺损伤及氧化应激相关信号通路Toll样受体（TLR）-4表达的影响。将6～8周龄、雌性SPF级BALB/c小鼠随机分为病毒感染致急性肺损伤(H9N2)组、H9N2+EGCG组、模拟感染（Mock）组、Mock+EGCG以及TLR-4抑制剂Eritoran5564(H9N2+E5564)对照组，观察各组小鼠肺组织病理学变化，测定肺湿/干重比；测定肺组织内MPO、T-SOD、抗HO•能力、MDA、IL-1β和TNF-α的含量；Western blot和RT-PCR方法检测肺内TLR-4 mRNA及蛋白质的表达。结果表明，H9N2组小鼠精神沉郁、呼吸困难、体重下降明显；肺组织学表现为肺泡壁水肿、炎性细胞浸润、出血为特征的弥漫性肺组织损伤。EGCG干预后，与H9N2组相比，EGCG治疗组小鼠临床症状较轻，一定程度上降低了死亡率，并明显延长小鼠存活时间(P<0.01)；其肺组织损伤程度较轻，肺湿/干重比极显著下降(P<0.01)；MPO和MDA的含量显著降低，与之相反T-SOD及抗HO•能力升高；同时，EGCG显著降低肺组织内IL-1β和TNF-α的含量。肺组织内TLR-4 mRNA以及蛋白表达显著降低； E5564呈现与EGCG相似的干预效果，显著降低TLR-4 mRNA以及蛋白表达，降低肺组织MPO、MDA、IL-1β和TNF-α的含量，减少T-SOD的消耗以及提高抗HO•能力。EGCG明显缓解小鼠肺损伤过程，其机制可能与其影响活性氧自由基的产生或清除进而显著降低TLR-4的表达有关，提示其在辅助预防和干预H9N2-SIV诱导的肺损伤方面具有潜在的应用前景。

Abstract:

The present research investigated effect of epigallocatechin gallate (EGCG) on toll-like receptor（TLR）-4 expression in lung injury induced by H9N2 swine influenza virus infection in mice.A total of 400 female BALB/c (SPF) mice were randomized into five groups and treated as follow：(1) H9N2 group，(2) H9N2+EGCG treatment group，(3) mock control，(4) mock+EGCG and (5) pharmacological inhibitor of TLR-4 Eritoran5564 (H9N2+E5564).The lung histopathology，lung water content，and MPO activity，OH• scavenging activity，the content of MDA and cytokines (IL-1β and TNF-α)，and the mRNA and protein expression of TLR-4 in the lung tissue were observed or investigated at 2，4，6，8，and 14 days after inoculation.The results showed：(1) H9N2 virus-infected mice presented depression，dyspnea and weight loss dramatically；Histopathologically，alveolar and interstital edema，hemorrhage and inflamatory cell infiltration were observed in H9N2-infected mice.(2) Compare to that of H9N2-inected mice，the EGCG treatment alleviated clinical signs and the histological lesion，prolonged survival time and decreased mortality (35% vs 65%，P<0.05).It also inhibited MPO activity，decreased MDA content，and increased T-SOD level and OH• scavenging activity.Moreover，the lung wet weight to dry weight ratio (P<0.05) and the content of IL-1β and TNF-α in lung tissue significantly decreased compare with that of H9N2-inected mice.(3) The EGCG treatment markedly down-regulated the levels of mRNA and protein of TLR-4 in the lungs of H9N2-infected mice (P<0.05).Similarly，the E5564 inhibited the expression of TLR-4 mRNA and protein in lung tissue of H9N2 virus infected mice；It also decreased dramatically the levels of MPO，MDA，IL-1β and TNF-α，but the levels of T-SOD and OH• scavenging activity were higher than H9N2 group.These data demonstrated that EGCG down-regulated remarkably the levels of mRNA and protein of TLR-4 and effected the produce the levels of MPO，MDA，IL-1β and TNF-α during H9N2 viral infection，thus supporting the use of EGCG for managing ALI induced by H9N2-SIV influenza in future.

中图分类号:

S855.3

徐彤，王存连，徐明举，张瑞华，刘宝剑，魏东，王国华. 表没食子儿茶素没食子酸酯对H9N2猪流感病毒诱导小鼠肺损伤中Toll样受体-4表达的影响[J]. 畜牧兽医学报, 2015, 46(8): 1438-1446.

XU Tong，WANG Cun-lian，XU Ming-ju，ZHANG Rui-hua，LIU Bao-jian，WEI Dong，WANG Guo-hua. Effect of Epigallocatechin Gallate on Toll-like Receptor-4 Expression in Mouse Model of Acute Lung Injury Induced by H9N2 Swine Influenza Virus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(8): 1438-1446.

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