金银花-连翘药对改善川崎病小鼠血管炎的作用及机制  

Study on the Efficacy of Lonicerae Japonicae Flos-Forsythiae Fructus on Ameliorating Vasculitis in Kawasaki Disease Mouse Model

作　　者：马锦[1] 唐孕佳 宋秀道 吕海涛[2] MA Jin;TANG Yunjia;SONG Xiudao;LYU Haitao(Department of Pharmacy,Children’s Hospital of Soochow University,Suzhou 215025 Jiangsu,China;Department of Cardiology,Children’s Hospital of Soochow University,Suzhou 215025 Jiangsu,China;Chinese Medicine Science and Technology Transformation Center,Suzhou Hospital of Traditional Chinese Medicine Affiliated to Nanjing University of Chinese Medicine,Suzhou 215009 Jiangsu,China)

机构地区：[1]苏州大学附属儿童医院临床药学室,江苏苏州215025 [2]苏州大学附属儿童医院心血管内科,江苏苏州215025 [3]南京中医药大学附属苏州市中医医院中医药科技转化中心,江苏苏州215009

出　　处：《中药新药与临床药理》2025年第3期349-356,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(82270529);苏州市科技局科研项目(SKY2023191,SKYD2023216);苏州市姑苏卫生人才计划项目(GSWS2024093)。

摘　　要：目的探讨金银花-连翘药对改善川崎病小鼠血管炎的作用及机制。方法将C57BL/6小鼠随机分为正常组(5只)、模型组(6只)和中药组(5只)。采用腹腔注射0.2 mL白色念珠菌水溶物(20 mg·mL^(-1))复制川崎病小鼠模型,每日1次,连续5 d。模型复制的同时,中药组小鼠给予金银花-连翘提取物(生药量2.6 g·kg^(-1))灌胃,正常组、模型组灌胃等量PBS。末次注射后,继续喂养14 d,然后取样进行后续实验。采用HE及EVG染色法观察心脏组织病理变化;免疫荧光法检测心脏组织中Ly6G、基质金属蛋白酶9(MMP9)表达水平;细胞因子检测试剂盒检测小鼠外周血中40种细胞因子的表达水平,并筛选差异表达细胞因子,进行GO功能及KEGG通路富集分析。结果(1)与正常组比较,模型组小鼠心脏血管内皮粗糙,结构不完整,细胞排列无序,血管及周围心肌组织可见炎性细胞广泛浸润;心脏血管壁弹力纤维排列紊乱,松散断裂;血管及周围心肌组织中可见大量中性粒细胞广泛浸润,Ly6G表达显著上调(P<0.01);心脏组织中MMP9蛋白表达显著上调(P<0.01)。与模型组比较,中药组小鼠心脏血管内皮结构较完整,炎性细胞浸润减轻;心脏血管壁弹力纤维较规整,未见明显断裂;心脏血管区域的中性粒细胞浸润明显减少,Ly6G表达显著下调(P<0.01);心脏组织中MMP9蛋白表达显著下调(P<0.01)。(2)与正常组比较,模型组有28个差异表达细胞因子,均显著上调(P_(adj)<0.05);与模型组比较,中药组有17个差异表达细胞因子,其中16个显著下调(P_(adj)<0.05),1个显著上调(P_(adj)<0.05)。有14个差异表达细胞因子为正常组vs模型组的上调基因,而在金银花-连翘干预后显著回调。差异表达细胞因子涉及白细胞迁移、细胞因子介导的信号通路、粒细胞趋化性等生物过程;涉及细胞因子活性、细胞因子受体结合、趋化因子活性等分子功能;以及涉及趋化因子信号通路、TNF信号�Objective To explore the effect and mechanism of Lonicerae Japonicae Flos-Forsythiae Fructus on improving vasculitis in Kawasaki disease mouse model.Method C57BL/6 mice were randomly divided into the normal group(n=5),model group(n=6)and Chinese medicine group(n=5).The Kawasaki disease mouse model was replicated by intraperitoneal injection of 0.2 mL Candida albicans water soluble fraction(20 mg·mL^(-1))once a day for 5 days.At the same time of model replication,the mice in the Chinese medicine group were given Lonicerae Japonicae Flos-Forsythiae Fructus extract(crude drug 2.6 g·kg^(-1))by gavage,and the normal group and the model group were given the same amount of PBS by gavage.After the last injection,the rats were fed for 14 days,and then sampled for subsequent experiments.HE and EVG staining were used to observe the pathological changes of heart tissue.The expression levels of Ly6 G and MMP9 in heart tissue were detected by immunofluorescence.The cytokine assay kit was used to detect the expression levels of 40 cytokines in peripheral blood of mice,and the differentially expressed cytokines were screened for GO function and KEGG pathway enrichment analysis.Results(1)Compared with the normal group,the cardiac vascular endothelium of the model group was rough and the structure was incomplete,the cells were disorderly arranged,and the inflammatory cells were widely infiltrated in the blood vessels and surrounding myocardial tissues.Cardiac vascular wall elastic fibers arranged in disorder,loose fracture;a large number of neutrophils were widely infiltrated in the blood vessels and surrounding myocardial tissues,and the expression of Ly6 G was significantly up-regulated(P<0.01).The protein expression of MMP9 in heart tissue was significantly upregulated(P<0.01).Compared with the model group,the cardiac vascular endothelial structure of the mice in the Chinese medicine group was more complete,and the infiltration of inflammatory cells was reduced;the elastic fibers of the cardiac vascular wall were relatively r

关 键 词：金银花-连翘药对 川崎病 血管炎 中性粒细胞 差异表达细胞因子 基质金属蛋白酶9 小鼠 

分 类 号：R285.5[医药卫生—中药学]