基于Caspase11-GSDMD-GSDMD-N通路探讨清瘟败毒饮对脓毒症小鼠肺损伤作用机制  

Exploring the Mechanism of Qingwen Baidu Drink on Lung Injury in Septic Mice Based on the Caspase11-GSDMD-GDMD-N Pathway

作　　者：赵裕沛[1] 白宇[2] 贺彬婵 庞薇[1] 刘梦云[1] 朱益敏[1] ZHAO Yupei;BAI Yu;HE Binchan;PANG Wei;LIU Mengyun;ZHU Yimin(The Second Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210017,China;School of Integrative Medicine,Nanjing University of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]南京中医药大学第二附属医院,江苏南京210017 [2]南京中医药大学中西医结合学院,江苏南京210023

出　　处：《南京中医药大学学报》2025年第3期333-340,共8页Journal of Nanjing University of Traditional Chinese Medicine

基　　金：国家自然科学基金青年科学基金项目(82100088);南京中医药大学自然科学基金(XZR2021057);南京中医药大学第二附属医院重点项目(SEZ202105)。

摘　　要：目的探究清瘟败毒饮治疗脓毒症诱导的肺损伤的作用机制。方法100只C57BL/6小鼠随机分为空白组、模型组、清瘟败毒饮低剂量组、清瘟败毒饮中剂量组、清瘟败毒饮高剂量组,每组20只。用HE染色检查肺组织的病理情况,ELISA法检测血清白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、趋化因子配体10(CXCL10)及血浆凝血因子Ⅲ(F3)的表达水平,qPCR检测肺组织中单核细胞趋化蛋白-1(MCP-1)、环氧化酶-2(COX-2)、干扰素-γ(IFN-γ)mRNA表达水平,血常规分析仪分析血浆中血小板(PLT)数量,免疫荧光分析法检测肺泡组织毛细血管内皮细胞的血管内皮钙黏素(VE-cadherin)、内皮黏附物连接标记物闭合蛋白5(CLDN5)以及周细胞标记物神经元胶原抗原2(NG2),Western blot检测小鼠肺组织中含半胱氨酸的天冬氨酸蛋白水解酶11(Caspase 11)、GSDMD、GSDMD-N的蛋白表达水平。结果与空白组相比,模型组小鼠肺组织呈现明显病理损伤变化,血清IL-1β、TNF-α、CXCL10水平和肺组织MCP-1、COX-2、IFN-γmRNA表达水平明显增高(P<0.01),血浆中PLT数量和F3含量明显降低(P<0.01);肺组织中VE-cadherin、CLDN5、NG2蛋白荧光表达明显增强(P<0.01),Caspase 11、GSDMD-N蛋白表达升高(P<0.01);与模型组相比,清瘟败毒饮各剂量组小鼠肺组织病理损伤均减轻,血清中IL-1β、TNF-α、CXCL10水平和肺组织中MCP-1、COX-2、IFN-γmRNA表达水平明显降低(P<0.05,P<0.01),血浆中PLT数量和F3含量均增高(P<0.05,P<0.01);肺组织中VE-cadherin、CLDN5、NG2蛋白荧光表达均减弱(P<0.05,P<0.01),Caspase11、GSDMD-N/GSDMD蛋白表达均降低(P<0.05,P<0.01)。结论清瘟败毒饮通过抑制GSDMD-N和Caspase 11的活化,减少炎症因子的释放,减少失血和血管屏障功能的损伤,从而对脓毒症引起的肺损伤起到一定的改善作用。OBJECTIVE To explore the mechanism of Qingwen Baidu Drink in treating sepsis-induced lung injury.METHODS One hundred C57BL/6 mice were randomly divided into blank group,model group,Qingwen Baidu Drink low-dose group,Qingwen Baidu Drink medium-dose group,and Qingwen Baidu Drink high-dose group,with 20 mice in each group.HE staining was used to examine the pathological changes of lung tissues.ELISA was used to detect the expression levels of serum interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),chemokine ligand 10(CXCL10)and plasma coagulation factorⅢ(F3).qPCR was used to detect the mRNA expression levels of monocyte chemoattractant protein-1(MCP-1),cyclooxygenase-2(COX-2)and interferon-γ(IFN-γ)in lung tissues.The number of platelets(PLT)in plasma was analyzed by routine blood analysis instrument.Immunofluorescence analysis was used to detect vascular endothelial cadherin(VE-cadherin),endothelial adhesion junction marker occludin 5(CLDN5)and pericyte marker neuronal collagen antigen 2(NG2)in alveolar capillary endothelial cells.Western blot was used to detect the protein expression levels of cysteine-containing aspartate proteinase 11(Caspase11),GSDMD and GSDMD-N in mouse lung tissues.RESULTS Compared with the blank group,the lung tissue of the mice in the model group showed obvious pathological damage.The levels of serum IL-1β,TNF-α,and CXCL10 and the mRNA expression levels of MCP-1,COX-2,and IFN-γin lung tissue were significantly increased(P<0.01),and the number of PLT and the content of F3 in plasma were significantly decreased(P<0.01).The fluorescence expression of VE-cadherin,CLDN5,and NG2 proteins in lung tissue was significantly enhanced(P<0.01),while the expression of Caspase11 and GSDMD-N proteins was increased(P<0.01).Compared with the model group,the pathological damage of the lung tissue of the mice in all doses of Qingwen Baidu Drink groups was alleviated,the levels of serum IL-1β,TNF-α,and CXCL10 and the mRNA expression levels of MCP-1,COX-2,and IFN-γin lung tissue were significantl

关 键 词：脓毒症 肺损伤 F3 Caspase11 GSDMD GSDMD-N 清瘟败毒饮 

分 类 号：R285.5[医药卫生—中药学]