注射用益气复脉(冻干)对脓毒症诱导小鼠急性肺损伤的改善作用研究  

Yiqifumai Lyophilized Injection Attenuates Sepsis-induced Acute Lung Injury in Mice

作　　者：潘雪薇 薛漓轩 张佳智 代玉洁 李芳[1,2] 张媛媛 寇俊萍[1,2] Pan Xuewei;Xue Lixuan;Zhang Jiazhi;Dai Yujie;Li Fang;Zhang Yuanyuan;Kou Junping(China Pharmaceutical University,Jiangsu,Nanjing 211198,China)

机构地区：[1]中国药科大学,江苏南京211198 [2]江苏省中药评价与转化重点实验室,江苏南京211198

出　　处：《中国中医急症》2021年第7期1129-1133,1168,共6页Journal of Emergency in Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81773971);中国药科大学“双一流”创新团队项目(CPU2018GF07)。

摘　　要：目的观察注射用益气复脉(冻干)通过调节血管内皮屏障功能对脓毒症所致小鼠急性肺损伤的保护作用及机制。方法C57BL/6J雄性小鼠随机分为假手术组、模型组、益气复脉(1、2、4 g/kg)组与地塞米松组。采用尾静脉注射的方式给予小鼠相应剂量的药物或等体积生理盐水后1 min内采用盲肠结扎穿孔术制备小鼠脓毒症模型。于手术后18 h,采用HE染色观察肺病理形态学变化;通过检测肺湿重干重比,肺泡灌洗液中白细胞、中性粒细胞和淋巴细胞数目,肺组织、血浆及BALF中髓过氧化物酶活力,探讨益气复脉对脓毒症小鼠肺组织的保护作用;用伊文思蓝渗漏法检测肺微血管渗漏情况;用蛋白免疫印迹方法观察TLR4、p-Src蛋白表达;采用免疫荧光法和蛋白免疫印迹检测VE-cadherin和p120-catenin的蛋白表达情况。结果与假手术组相比,模型组小鼠肺病理形态损伤及肺部水肿严重,BALF中白细胞、中性粒细胞和淋巴细胞数目显著升高,肺组织、血浆及BALF中MPO活力显著提高,肺微血管伊文思蓝渗漏增加,肺组织中TLR4和p-Src蛋白表达显著升高,而VE-cadherin和p120-catenin的蛋白表达显著降低(P<0.05或P<0.01)。益气复脉可明显改善模型小鼠肺组织损伤及肺水肿,缓解模型小鼠肺组织病理学改变,减轻肺部水肿,减少BALF中白细胞、中性粒细胞、淋巴细胞数目,抑制血浆、肺组织和BALF中MPO活力的增加,减轻肺微血管伊文思蓝渗漏,其中2 g/kg益气复脉为最优给药剂量,可以抑制TLR4表达的升高及Src磷酸化,恢复VE-cadherin和p120-catenin的表达,恢复细胞间黏附连接(P<0.05或P<0.01)。结论益气复脉可能通过调节TLR4/Src/VE-cadherin/p120-catenin信号途径,保护血管内皮屏障功能,改善脓毒症诱导的小鼠急性肺损伤。Objective:To observe the therapeutic effect of Yiqifumai lyophilized injection(YQFM)on sepsis-induced acute lung injury(ALI)in mice by regulating the vascular endothelial barrier function and to investigate the underlying mechanism.Methods:Male C57 BL/6 J mice were randomly divided into the sham group,model group,YQFM groups(1 g/kg,2 g/kg and 4 g/kg)and Dexamethasone(5 mg/kg)group.Mice were injected with the corresponding dose of drugs or equal volume of normal saline by tail intravenous injection to establish sepsis model by cecal ligation and puncture within 1 min.At 18 h after surgery,HE staining was used to observe lung pathomorphology changes;lung wet weight to dry weight ratio was measured,the number of leukocytes,neutrophils and lymphocytes in alveolar lavage fluid,lung tissue,plasma and BALF Oxidase activity were used to investigate the protective effect of YQFM on lung tissue of septic mice;Evans blue leakage was used to detect pulmonary microvascular leakage;Western blotting was used to detect the TLR4,p-Src protein expression level;immunofluorescence and Western blotting were used to observe the protein expression of VE-cadherin and p120-catenin.Results:Compared with the sham group,the lung pathological damage and pulmonary edema were severe in the model group;the number of white blood cells,neutrophils and lymphocytes in BALF were significantly increased,and the activity of MPO in lung tissue,plasma and BALF was significantly increased.The leakage of pulmonary microvascular Evans blue increased,the expressions of TLR4 and p-Src protein in lung tissue were significantly increased,while the protein expressions of VE-cadherin and p120-catenin were significantly decreased(P<0.05 or P<0.01).YQFM could significantly improve lung tissue damage and pulmonary edema in the model group,relieved pathological changes of lung tissue,reduced lung edema,the number of white blood cells,neutrophils,and lymphocytes in BALF,inhibited the increase of MPO activity in plasma,lung tissue and BALF,and reduce the Evans blue le

关 键 词：脓毒症 急性肺损伤 注射用益气复脉(冻干) 内皮通透性 盲肠结扎穿孔术 

分 类 号：R285.5[医药卫生—中药学]