蒲地蓝消炎口服液对甲型流感病毒感染小鼠肺上皮屏障的保护作用及分子机制  

Protective effect and molecular mechanism of Pudilan anti-inflammatory oral liquid on lung injury in mice infected with the influenza A virus

作　　者：万驰恒 杨道文[1,2,3] 孔明慧 卢雅梦[2,4] 邓博 李红艳 左文婷 何迪[2,4,7] 黄一皎 吴凯薇 袁英瑛 邹晓辉 李得民 王在 WAN Chi-heng;YANG Dao-wen;KONG Ming-hui(Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学中日友好医院临床医学院,北京100029 [2]国家呼吸医学中心,呼吸和共病全国重点实验室,国家呼吸疾病临床研究中心,中国医学科学院呼吸病学研究院,中日友好医院呼吸中心,北京100029 [3]中日友好医院中医肺病一部,北京100029 [4]中日友好医院呼吸与危重症医学科,北京100029 [5]中日友好临床医学研究所,北京100029 [6]北京协和医学院,中国医学科学院,北京100730 [7]首都医科大学中日友好临床医学院,北京100029 [8]清华大学基础医学院,北京100084 [9]北京大学中日友好临床医学院,北京100029

出　　处：《中日友好医院学报》2024年第6期338-344,F0002,F0003,共9页Journal of China-Japan Friendship Hospital

基　　金：中医药创新团队及人才支持计划;国家中医药多学科交叉创新团队项目(ZYYCXTD-D-202208);国家重点研发计划(2022YFC3500801)。

摘　　要：目的:探究蒲地蓝消炎口服液对甲型流感病毒感染小鼠肺损伤的保护作用及分子机制。方法:将40只C57BL/6J雄性小鼠随机分为4组,每组10只,分别是对照组、模型组、蒲地蓝消炎口服液低剂量和高剂量组。模型组为H1N1/PR8病毒滴鼻造模。蒲地蓝干预组为造模后连续5d灌胃给药。感染后第5d取血检测,对肺组织进行HE染色和免疫组化染色分析,并通过转录组测序分析肺组织基因表达差异,荧光定量RT-PCR进行验证。结果:蒲地蓝消炎口服液能够减轻肺部炎症及组织损伤;可降低甲流小鼠的中性粒细胞百分数、血小板计数,升高甲流小鼠的淋巴细胞百分比,模型组与低剂量组比较差异有统计学意义(均P<0.05);并维持支气管上皮细胞形态。转录组研究发现,蒲地蓝对于流感病毒感染后部分细胞骨架、细胞连接、细胞外基质及上皮细胞分泌相关基因的表达变化具有逆转作用,荧光定量RT-PCR结果,高剂量组对于Cldn3、Col4a3和Mybpc2基因表达具有调节作用(均P<0.05)。结论:蒲地蓝可能通过调节免疫炎症反应,维持肺上皮细胞基因表达,改善上皮屏障完整性,减轻流感病毒感染引起的小鼠肺损伤。Objective:To investigate the protective effect and molecular mechanism of Pudilan anti-inflammatory oral liquid on lung injury in mice infected with the influenza A virus(IAV).Methods:Forty male C57BL/6J mice were randomly divided into four groups,with 10 mice in each group:control group,model group,lowdose and high-dose Pudilan anti-inflammatory oral liquid group.The model group was created by intranasal instillation of the H1N1/PR8 virus.The Pudilan intervention groups received oral administration for 5 consecutive days after modeling.Results:Pudilan anti-inflammatory oral liquid can reduce lung inflammation and tissue damage caused by IAV infection.It can reduce the percentage of neutrophils and platelet count in the mice infected with IAV,and increase the percentage of lymphocytes(all P<0.05);and maintain the morphology of bronchial epithelial cells.Transcriptome analysis revealed that Pudilan has a reversing effect on the expression changes of genes related to the cell cytoskeleton,cell junctions,extracellular matrix,and epithelial cell secretion following influenza virus infection.Through fluorescence quantitative RT-PCR verification,it was found that the high-dose group has a regulatory effect on the gene expression of Cldn3,Col4a3,and Mybpc2(all P<0.05).Conclusion:Pudilan may regulate the immune inflammatory response,maintain gene expression in lung epithelial cells,improve epithelial barrier integrity,and alleviate lung injury caused by influenza virus infection in mice.

关 键 词：蒲地蓝消炎口服液 甲型流感病毒 肺炎 肺上皮屏障 细胞连接 小鼠 

分 类 号：R373.1-3[医药卫生—病原生物学]