乌药调控p38MAPK/ERK通路对脂多糖诱导ARDS小鼠的肺保护作用  

Original Article Protective effect of Lindera aggregata on lipopolysaccharide-induced mice acute respiratory distress syndrome by regulating p38MAPK/ERK pathway

作　　者：陆明峰 范璐 徐敏 嵇玲 徐继扬 Lu Mingfeng;Fan Lu;Xu Min;Ji Ling;Xu Jiyang(Department of Emergency Medicine,Clinical Medical College,Yangzhou University,Yangzhou 225001,Jiangsu,China)

机构地区：[1]扬州大学临床医学院急诊科,江苏扬州225001

出　　处：《中华危重病急救医学》2022年第9期947-951,共5页Chinese Critical Care Medicine

基　　金：江苏省中医药局科技项目(YB2017076);江苏省扬州市科技计划项目(YZ2020091)。

摘　　要：目的探讨乌药对脂多糖(LPS)诱导急性呼吸窘迫综合征(ARDS)小鼠的肺保护作用及可能机制。方法将40只C57BL/6小鼠随机分为假手术组、ARDS模型组、乌药低剂量组、乌药高剂量组,每组10只。经气管注射5 mg/kg的LPS制备小鼠ARDS模型;乌药低剂量组和高剂量组分别给予乌药提取物1 g/kg和5 g/kg每日1次灌胃,ARDS模型组以等量生理盐水灌胃;假手术组不进行任何处理。制模前预给药3 d,制模后继续给药2 d并处死动物,取支气管肺泡灌洗液(BALF)和肺组织。光镜下观察各组小鼠肺组织病理学改变,测肺湿/干质量比值(W/D);采用酶联免疫吸附试验(ELISA)检测小鼠血清及BALF中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)含量;采用流式细胞术检测BALF中巨噬细胞表面分子CD40表达率;采用蛋白质免疫印迹试验(Western blotting)检测肺组织p38和细胞外信号调节激酶1/2(ERK1/2)蛋白磷酸化水平变化。结果肺组织病理学显示,乌药高剂量组光镜下肺泡结构破坏、肺泡间隔增厚、炎症细胞浸润,但病理变化均较ARDS模型组减轻,而乌药低剂量组ARDS病理特征较ARDS模型组无明显改变。与假手术组比较,ARDS模型组肺W/D比值明显升高,血清及BALF中TNF-α和IL-6含量明显升高,BALF中巨噬细胞CD40表达率明显增加,肺组织p38和ERK1/2蛋白磷酸化水平明显升高。用乌药干预后,低剂量组肺组织W/D比值、血清及BALF中TNF-α和IL-6水平、BALF中巨噬细胞CD40表达率、肺组织p38和ERK1/2蛋白磷酸化水平均较ARDS模型组无明显变化;而乌药高剂量组上述指标均明显低于ARDS模型组和乌药低剂量组〔W/D比值:5.70±0.19比6.20±0.31、6.01±0.17;血清TNF-α(ng/L):83.63±15.04比111.75±18.45、108.12±13.98;血清IL-6(ng/L):111.38±8.75比244.13±26.85、227.50±9.37;BALF中TNF-α(ng/L):36.25±2.82比51.13±5.44、47.50±5.78;BALF中IL-6(ng/L):35.63±2.20比49.63±4.90、46.38±3.50;CD40表达率(%):23.28±2.45比30.32±Objective To explore the protective effect of Lindera aggregata on acute respiratory distress syndrome(ARDS)induced by lipopolysaccharide(LPS)in mice and its possible mechanism.Methods Forty C57BL/6 mice were randomly divided into sham operation group,ARDS model group,low-dose Lindera aggregata(L-LA)group and high-dose Lindera aggregata(H-LA)group,with 10 mice in each group.ARDS model was established by injecting 5 mg/kg LPS through the trachea.The L-LA group and H-LA group were orally administrated 1 g/kg and 5 g/kg of the Lindera aggregate extract once a day,respectively,while the ARDS model group was given the same volume of normal saline,the sham group received no treatment.The Lindera aggregata was preadministered for 3 days before modeling,and continued for 2 days after modeling,then the animals were sacrificed,and bronchoalveolar lavage fluid(BALF)and lung tissue were collected.The pathological changes of lung tissue in each group of mice were observed under the microscope and the wet/dry weight ratio(W/D)of the lung were measured.Enzyme linked immunosorbent assay(ELISA)was used to examine the levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in mice serum and BALF,and flow cytometry was used to detect the expression rate of CD40 on the surface of BALF macrophages.The phosphorylation levels of p38 and extracellular signal-regulated protein kinase 1/2(ERK1/2)proteins in lung tissue were measured by Western blotting.Results Lung histopathology under light microscope showed that the damage of alveolar structure,thickening of alveolar septum and infiltration of inflammatory cells in the H-LA group were less severe than those in the ARDS model group,while the pathological characteristics of ARDS in the L-LA group were not significantly different from those in the ARDS model group.Compared with the sham operation group,the lung W/D ratio,TNF-αand IL-6 protein contents in serum and BALF,BALF macrophage CD40 expression rate and lung tissue p38 and ERK1/2 protein phosphorylation levels were signific

关 键 词：乌药 急性呼吸窘迫综合征 炎症信号通路 P38丝裂素活化蛋白激酶 细胞外信号调节蛋白激酶 

分 类 号：R285.5[医药卫生—中药学]