黄芪甲苷调控CD45 PTPase介导抗脓毒症免疫机制的研究  

Anti-Sepsis Immunological Mechanism of Astragaloside IV through Activating CD45 PTPase

作　　者：杨海浩 应赛 顾茜兰 邹楠婷 吴招 张春菲 张浩洪 万春平 YANG Haihao;YING Sai;GU Qianlan;ZOU Nanting;WU Zhao;ZHANG Chunfei;ZHANG Haohong;WAN Chunping(Yunnan University ofChinese Medicine,Kunming 650500,China;Yunnan College of Business Management,Kunming 650106,China;Zhengxiong Country Hospital of traditional medicine,Zhaotong 657200,China)

机构地区：[1]云南中医药大学,云南昆明650500 [2]云南经济管理学院,云南昆明650106 [3]镇雄县中医医院,云南昭通657200

出　　处：《云南中医药大学学报》2024年第1期57-61,78,共6页Journal of Yunnan University of Chinese Medicine

基　　金：国家自然科学基金项目(81960745)。

摘　　要：目的研究黄芪甲苷(Astragaloside IV,ASI IV)抗脓毒症效应及免疫学机制,为黄芪临床应用脓毒症治疗提供科学依据。方法采用盲肠结扎穿刺术(Cecal ligation and puncture,CLP)构建脓毒症小鼠模型,随机分为假手术组、模型组、黄芪甲苷组、Anti-CD45单抗组、Anti-CD45单抗+黄芪甲苷组。H&E染色检测肺组织病理损伤;流式细胞术检测脾脏Th1细胞表达水平;实时荧光定量PCR(Real-time PCR)检测Th1细胞相关基因(IL-2、Tbet、STAT1和STAT4)mRNA的表达。结果与模型组比较,黄芪甲苷干预显著提高脓毒症模型小鼠生存率,减轻模型小鼠肺组织病理损伤。CD45抗体干预后,阻断了黄芪甲苷提高生存率和减轻肺组织病理损伤的作用。机制研究结果表明,黄芪皂苷显著增加模型小鼠Th1细胞(CD4+IFN-γ+)比例,上调Th1细胞相关基因(IL-2、T-bet、STAT1、STAT4)mRNA表达,CD45抗体干预后,减少了黄芪甲苷促脾淋巴细胞Th1细胞比例,阻断黄芪皂苷促Th1细胞相关基因。结论黄芪甲苷通过调控CD45分子,提高Th1细胞介导的免疫反应,介导抗脓毒症效应。Objective The aim of this study was designed to investigate the effect of Astragaloside IV on sepsis and its immunological mechanism.This study will provide a basis for the theoretical foundation of anti-sepsis immunotherapy of Astragalus.Methods CLP(Cecal ligation and puncture)model was established to investigated the anti-septic potential and reveal its underlying mechanisms,the mice with CLP were divide into five groups including sham group,model group,Astragaloside IV group,Anti-CD45 Ab group and Astragaloside II+Anti-CD45 Ab group.Lung injury in sepsis mice was assessed by H&E staining.The percentage of Th1 cells(CD4+IFN-γ+)were detected by flow cytometry.The mRNA expression of Th1 cytokine and transcript factor T-bet were examined by q-PCR analysis.Results Compared with model group,treatment of Astragaloside IV can markedly improve the survival rate and reduce inflammatory lung injury in sepsis mice.However,anti-CD45 Ab treatment intensely blocked anti-septic effect including the enhanced survival rate and lung injure,which induced by Astragaloside IV.Furthermore,expression of Th1 cells(CD4+IFN-γ+)and Th1 cells related gene(IL-2、T-bet、STAT1、STAT4)mRNA expression were markedly increased in Astragaloside IV group comparison with model group.Whereas,the percentage of Th1 cells and Th1 cells related gene were significantly decreased in Astragaloside IV+Anti-CD45 Ab group.Conclusion Astragaloside IV might promote Th1 cell-mediated immune response in sepsis through CD45 protein tyrosine phosphatase activity.This mechanism will provide a basis for the clinical application of Astragalus in treating sepsis.

关 键 词：脓毒症 黄芪甲苷 CD45蛋白酪氨酸磷酸酶 Th1细胞 盲肠结扎穿孔术 

分 类 号：R285.5[医药卫生—中药学]