加味麻杏石甘汤治疗铜绿假单胞菌肺炎大鼠疗效及对NADPH氧化酶-依赖性NETs的调控作用  

The Efficacy of Modified Maxing Shigan Decoction(加味麻杏石甘汤)in Treating Pseudomonas Aeruginosa Pneumonia in Rats and Its Regulatory Effection on NADPH Oxidase-dependent NETs

作　　者：刘奉云 李修齐 王睦天 赵启亮[1,2] 刘凯 尹延重[1,2] 孙宏源 刘旻 LIU Fengyun;LI Xiuqi;WANG Mutian;ZHAO Qiliang;LIU Kai;YIN Yanzhong;SUN Hongyuan;LIU Min(First Teaching Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300381,China;National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion,Tianjin 300381,China;Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China)

机构地区：[1]天津中医药大学第一附属医院,天津300381 [2]国家中医针灸临床医学研究中心,天津300381 [3]天津中医药大学,天津301617

出　　处：《辽宁中医药大学学报》2025年第5期17-21,共5页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：国家自然科学基金(82205185)。

摘　　要：目的观察加味麻杏石甘汤对铜绿假单胞菌(PA)肺炎大鼠的疗效及对NADPH氧化酶-依赖性NETs的调控作用。方法选取雄性SD大鼠160只,随机分为空白组32只和造模组128只。造模组采用气管滴注法建立PA肺炎模型。建模后,所有大鼠随机分为模型组、中药组、模型+肽酰基精氨酸脱亚胺酶4(PAD4)抑制剂组和中药+PAD4抑制剂组,每组32只。造模前,模型+PAD4抑制剂组和中药+PAD4抑制剂组予腹腔注射PAD4抑制剂(Cl-脒)3 d;造模后,空白组、模型组和模型+PAD4抑制剂组予0.9%氯化钠溶液灌胃,中药组和中药+PAD4抑制剂组予等量中药浓缩煎剂灌胃。记录并评估所有大鼠一般状态,分4个时点(造模后6 h、12 h、24 h、72 h)取肺组织,每个时点每组取8只;采用HE染色观察病理改变;采用菌落计数法和酶联免疫法检测PA菌落数,瓜氨酸化组蛋白H3(Cit H3)、弹性蛋白酶(NE)-DNA复合物、PAD4和NADPH氧化酶的量。结果(1)各组大鼠一般状态和肺组织病理评分均以中药组最低,中药+PAD4抑制剂组次之,模型+PAD4抑制剂组最高。(2)各组内PA菌落计数逐渐减低,差异有统计学意义(P<0.05);中药组在6 h和72 h时菌落数显著低于模型+PAD4抑制剂组和模型组(P<0.05)。(3)NETs标记物(Cit H3和NE-DNA)的量,中药组和中药+PAD4抑制剂组内均呈上升再下降趋势;二者在6 h和12 h,中药组显著高于空白组和模型+PAD4抑制剂组(P<0.05);72 h,中药组Cit H3水平显著低于模型组(P<0.05)。(4)组内PAD4和NADPH氧化酶的量在中药组和中药+PAD4抑制剂组差异均无统计学意义(P>0.05),前者呈下降趋势,后者为上升再下降趋势;12 h,与模型+PAD4抑制剂组比较,中药组二者水平均显著增高(P<0.05);72 h,与模型组比较,中药组二者水平均显著降低(P<0.05)。结论加味麻杏石甘汤治疗PA肺炎大鼠疗效显著,其可通过NADPH氧化酶-依赖性途径双向调控肺组织NETs;肺炎形成期促进NETs,增强PMNs杀菌效应,消退期Objective To observe the therapeutic effect of Modified Maxing Shigan Decoction(加味麻杏石甘汤)on Pseudomonas aeruginosa(PA)pneumonia in rats and its regulatory effect on NADPH oxidase-dependent NETs.Methods 160 male SD rats were selected and randomly divided into a blank group with 32 rats and a modeling group with 128 rats.The modeling group established a PA pneumonia model using tracheal instillation.After modeling,all rats were randomly divided into a model group,Chinese medicine group,model+PAD4 inhibitor group,and Chinese medicine+PAD4 inhibitor group,with 32 rats in each group.Before modeling,the model+PAD4 inhibitor group and Chinese medicine+PAD4 inhibitor group were injected with the PAD4 inhibitor(Cl-amidine)at a dose of 12.5 mg/kg for 3 days.After modeling,the blank group,model group and model+PAD4 inhibitor group were given intragastric saline,while the Chinese medicine group and Chinese medicine+PAD4 inhibitor group were given an equivalent amount of concentrated decoction.The general condition of all rats was recorded and evaluated,and lung tissue samples were taken out at four time points(6,12,24,72 h after modeling),with 8 rats per group per time point.Pathological changes were observed using HE staining,and the colony count method and enzyme-linked immunosorbent assay were used to detect the colony counts of PA in lung tissue,the levels citrullinated histone H3(CitH3),neutrophil elastase(NE)-DNA complexes,PAD4 and NADPH oxidase.Results(1)The general condition and lung tissue pathology scores of rats in each group were lowest in the Chinese medicine group,followed by those in the Chinese medicine+PAD4 inhibitor group,and highest in the model+PAD4 inhibitor group.(2)The colony counts of PA gradually decreased in each group,with significant differences(P<0.05);the colony counts in the Chinese medicine group at 6 h and 72 h were significantly lower than those in the model+PAD4 inhibitor group and model group(P<0.05).(3)The levels of NETs markers(CitH3 and NE-DNA)in the Chinese medicine group

关 键 词：加味麻杏石甘汤 铜绿假单胞菌 肺炎 中性粒细胞胞外陷阱 NADPH氧化酶 

分 类 号：R256.1[医药卫生—中医内科学]