黄芩苷对自噬介导H6N6禽流感病毒致病损伤的作用  

Effect of Baicalin on Autophagy Mediated Pathogenic Injury of H6N6 Avian Influenza Virus

作　　者：杨鑫[1,2] 单春兰 杨剑 宋旭琴 徐浩 欧德渊 YANG Xin;SHAN Chun-lan;YANG Jian;SONG Xu-qin;XU Hao;OU De-yuan(College of Animal Science,Guizhou University,Guiyang,Guizhou,550025,China;Guizhou Provincial Key Laboratory of Animal Diseases and Veterinary Public Health,Guiyang,Guizhou,550025,China)

机构地区：[1]贵州大学动物科学学院,贵州贵阳550025 [2]贵州省动物疫病与兽医公共卫生重点实验室,贵州贵阳550025

出　　处：《动物医学进展》2023年第9期55-62,共8页Progress In Veterinary Medicine

基　　金：贵州省科学技术基金项目(黔科合基础[2020]1Y136)。

摘　　要：探究黄芩苷对H6N6亚型禽流感病毒诱导小鼠肺脏致病损伤的作用。选用40只昆明小鼠随机分为4组,即对照组、模型组、黄芩苷治疗组、3-MA抑制剂组。通过HE染色观察肺组织病理形态学变化,ELISA法检测血清中IL-1β、IL-2、IL-6及TNF-α的含量,透射电镜检测小鼠肺脏中自噬体的水平,Western blot检测肺组织LC3、Beclin-1及ATG7蛋白表达水平,免疫荧光法检测肺组织LC3蛋白表达水平,免疫组化法检测肺组织LC3、Beclin-1及ATG5蛋白表达水平,RT-qPCR检测肺组织自噬关键基因LC3、Beclin-1、ATG3、ATG5、ATG7及ATG12 mRNA转录水平。结果显示,经黄芩苷治疗后能显著缓解H6N6亚型禽流感病毒对小鼠肺组织的损伤,血清中IL-1β、IL-2、IL-6及TNF-α含量显著下降(P<0.01),肺组织中自噬体数量减少,LC3、Beclin-1、ATG5及ATG7蛋白表达水平显著下降(P<0.01),肺组织中LC3、Beclin-1、ATG3、ATG5、ATG7及ATG12 mRNA转录水平显著下降(P<0.01)。结果表明,黄芩苷能减轻H6N6亚型禽流感病毒诱导小鼠肺脏炎症损伤,同时下调自噬关键因子及自噬体的水平,提示其抗小鼠肺损伤机制可能与调控细胞自噬有关。To investigate the effect of baicalin on lung injury induced by H6N6 avian influenza virus in mice,forty Kunming mice were randomly divided into 4 groups:control group,model group,baicalin treatment group and 3-MA inhibitor group.The histopathological lesions of lung tissues were observed by HE staining.The levels of IL-1β,IL-2,IL-6 and TNF-αin sera were detected by ELISA.The level of autophagosome in lungs of mice was detected by transmission electron microscopy.The protein expressions of LC3,Beclin-1 and ATG7 in lung tissue were detected by Western blot.Immunofluorescence was used to detect the expression of LC3 protein in lung tissue.The protein expression levels of LC3,Beclin-1 and ATG5 in lung tissue were detected by immunohistochemistry.RT-qPCR was used to detect the mRNA transcription levels of autophagy key genes LC3,Beclin-1,ATG3,ATG5,ATG7 and ATG12.The results showed that baicalin could significantly alleviate the injury of lung tissue induced by H6N6 avian influenza virus in mice.Serum levels of IL-1β,IL-2,IL-6 and TNF-αwere significantly decreased(P<0.01);The number of autophagosomes in lung tissue decreased;The protein expression levels of LC3,Beclin-1,ATG5 and ATG7 were significantly decreased(P<0.01);The mRNA transcription levels of LC3,Beclin-1,ATG3,ATG5,ATG7 and ATG12 in lung tissue decreased significantly(P<0.01).The results showed that baicalin could reduce lung inflammatory injury induced by H6N6 avian influenza virus in mice,and down-regulate the levels of autophagy key factors and autophagosomes,suggesting that the mechanism of baicalin against lung injury in mice may be related to the regulation of autophagy.

关 键 词：黄芩苷 H6N6亚型禽流感病毒 自噬 炎症 

分 类 号：S852.659.5[农业科学—基础兽医学]