宣肺败毒方调控TLR4/NLRP3/Caspase1信号通路抗小鼠急性肺损伤的作用机制研究  

Mechanism of Xuanfei Baidu Decoction to improve ALI mice by regulating TLR4/NLRP3/Caspase1 signaling pathway

作　　者：邱先哲 黄晨瑶 李彩霞[2] 张淑坤[2] 崔立华[2] 李雯雯 刘国敬 齐育麟 张晗[1,3] 李琳[1,3] 李玉红[1,3] QIU Xianzhe;HUANG Chenyao;LI Caixia;ZHANG Shukun;CUI Lihua;LI Wenwen;LIU Guojing;QI Yulin;ZHANG Han;LI Lin;LI Yuhong(Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;Tianjin Nankai Hospital,Tianjin 300193,China;Tianjin University of Traditional Chinese Medicine,State Key Laboratory of Component-based Chinese Medicine,Tianjin 301617,China)

机构地区：[1]天津中医药大学中医药研究院,天津301617 [2]天津市南开医院,天津300193 [3]天津中医药大学组分中药国家重点实验室,天津301617

出　　处：《天津中医药大学学报》2024年第11期992-999,共8页Journal of Tianjin University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(82141201);国家重点研发计划(2020YFA0708004,2021YFE0200300)。

摘　　要：[目的]基于Toll样受体4(TLR4)/核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)/半胱氨酸蛋白酶-1(Caspase1)信号通路研究宣肺败毒方(XFBD)对IgG免疫复合物(IgG-IC)诱导小鼠急性肺损伤(ALI)的药效作用及机制研究。[方法]采用IgG-IC诱导小鼠构建ALI模型,灌胃XFBD 4 g生药/kg和8 g生药/kg,单次给药;苏木素-伊红染色法(HE)观察肺组织病理变化;乳酸脱氢酶(LDH)试剂盒测定细胞活力;荧光定量PCR法检测小鼠肺组织细胞因子白细胞介素-6(IL-6)、白细胞介素-18(IL-18)、白细胞介素-1β(IL-1β)、单核细胞趋化蛋白-1(MCP-1)以及TLR4/NLRP3/Caspase1信号通路相关因子mRNA表达量;免疫组化法检测小鼠肺组织TLR4、NLRP3和消皮素D(GSDMD)蛋白表达;蛋白免疫印迹法检测小鼠肺组织闭合蛋白(Occludin)、闭锁连接蛋白1(ZO-1)以及TLR4/NLRP3/Caspase1信号通路相关因子蛋白表达水平。[结果]与正常组相比,模型组小鼠肺组织炎性细胞浸润,肺泡间隙增厚,肺组织病理损伤评分和肺组织脏器系数显著升高,LDH及细胞因子表达明显升高,Occludin和ZO-1表达下调,TLR4/NLRP3/Caspase1信号通路相关因子mRNA和蛋白水平显著升高;与模型组相比,XFBD显著缓解急性肺损伤小鼠肺组织炎症细胞浸润,降低细胞因子表达,上调Occludin和ZO-1,显著下调TLR4/NLRP3/Caspase1信号通路相关因子mRNA和蛋白水平。[结论]XFBD可通过调控TLR4/NLRP3/Caspase1信号通路发挥抗小鼠ALI的作用。[Objective]Pharmacodynamic effects and mechanism of Xuanfei Baidu Decoction(XFBD)on IgG-IC-induced acute lung injury(ALI)in mice based on the Toll-like receptor 4(TLR4)/NOD-like receptor family pyrin domain containing 3(NLRP3)/Cysteine aspartate protein hydrolase-1(Caspase1)signaling pathway.[Methods]An ALI model was constructed using IgG-IC-induced mice,and XFBD 4 g raw/kg and 8 g raw/kg were administered by gavage in a single dose;pathological changes in lung tissue were observed by hematoxylin-eosin staining(HE).Cell viability was determined by lactate dehydrogenase(LDH)kit.And the fluorescence quantitative PCR was performed to detect the cytokines that include interleukin-6(IL-6),interleukin-18(IL-18),interleukin-1β(IL-1β)and monocyte chemotactic protein-1(MCP-1)in mouse lung tissue,and TLR4/NLRP3/Caspase1 signaling pathway related factors mRNA expression.Immunohistochemistry to detect the expression of TLR4,NLRP3,and Gasdermin-D(GSDMD)proteins in mouse lung tissues,and western blotting to detect the protein expression of Occludin,(Zonula occluding-1)ZO-1 and TLR4/NLRP3/Caspase1 signaling pathway related factors were detected by immunohistochemistry.[Results]Compared with the control group,mice in the model group had lung tissue inflammatory cell infiltration,alveolar interstitial thickening,significantly higher lung histopathological injury scores and lung tissue organ coefficients,exceedingly higher expression of LDH and cytokines,down-regulated expression of Occludin and ZO-1,and extraordinary higher mRNA and protein levels of factors related to the TLR4/NLRP3/Caspase1 signaling pathway.Compared with the model group,XFBD significantly alleviated inflammatory cell infiltration in lung tissues of mice with acute lung injury,reduced cytokine expression,up-regulated Occludin and ZO-1,and greatly down-regulated mRNA and protein levels of factors related to the TLR4/NLRP3/Caspase1 signaling pathway.[Conclusion]XFBD can exert anti-ALI effects in mice by modulating the TLR4/NLRP3/Caspase1 signaling pathway.

关 键 词：宣肺败毒方 急性肺损伤 TLR4/NLRP3/Caspase1信号通路 炎症 

分 类 号：R285.5[医药卫生—中药学]