益气化瘀解毒方对急性呼吸窘迫综合征大鼠肺损伤及中性粒细胞粘附分子的作用研究

Study on the effect of Yiqi Huayu Jiedu Formula on lung injury and neutrophil adhesion molecules in rats with acute respiratory distress syndrome

目的 观察益气化瘀解毒方及拆方对脂多糖(lipopolysaccharide, LPS)诱导的急性呼吸窘迫综合征(acute respiratory distress syndrome, ARDS)模型大鼠的保护作用及中性粒细胞粘附分子的影响。方法 将36只SD大鼠随机分为对照组、模型组、益气清热组、益气化瘀组、益气逐饮组及中药全方组,每组6只。益气清热组、益气化瘀组、益气逐饮组、中药全方组给予对应中药浸膏12.2 g/(kg·d)连续灌胃7天,其余组均以等量蒸馏水灌胃。尾静脉注射LPS建立ARDS大鼠模型,造模后16小时麻醉处死各组动物进行取材。苏木精—伊红染色观察肺组织病理变化,采用酶联免疫吸附法法检测肺匀浆及肺泡灌洗液中Rap1b、淋巴细胞功能相关抗原1(leukocyte function-associated antigen-1,LFA-1)、巨噬细胞表面抗原-1(macrophage surface antigen-1,MAC-1)的表达水平,蛋白免疫印迹法检测肺组织Rap1b的表达水平。结果 与对照组相比,模型组大鼠Rap1b、LFA-1及MAC-1蛋白表达均有所上调。与模型组相比,益气化瘀解毒方及拆方可明显减轻ARDS模型大鼠肺损伤,显著下调肺泡灌洗液中Rap1b、MAC-1及肺匀浆中LFA-1蛋白表达。结论 益气化瘀解毒方对LPS诱导的ARDS模型大鼠具有保护作用,可能与其减少Rap1b、LFA-1和MAC-1蛋白表达水平,降低中性粒细胞粘附活性有关。

Objective To observe the protective effects of the formula of Yiqi Huayu Jiedu Formula and the disassembling formula on lipopolysaccharide(LPS)-induced acute respiratory distress syndrome(ARDS)model rats and the effects of neutrophil adhesion molecules.Methods Thirty-six SD rats were randomly divided into a control group,a model group,a yiqi qingre group,a yiqi huayu group,a yiqi zhuyin group and a whole prescription group of traditional Chinese medicine,with six rats in each group.In the yiqi qingre group,the yiqi huayu group,the yiqi zhuyin group,and the whole formula group,12.2 g/(kg·d)of the corresponding Chinese herbal medicine extract was given by gavage for 7 days,and the rest of the groups were gavaged with an equal amount of distilled water.LPS was injected into the tail vein to establish the ARDS rat model,and the animals in each group were anaesthetised and executed 16 hours after modelling for sampling.Hematoxylin-eosin staining was used to observe the pathological changes in lung tissue,and enzyme-linked immunosorbent assay was used to detect the levels of Rap1b,lymphocyte function-associated antigen-1(LFA-1),macrophage surface antigen-1(LFA-1,macrophage surface antigen-1(MAC-1),and Rap1b in lung tissue by western-blot.Results Compared with the control group,the protein expression of Rap1b,LFA-1 and MAC-1 was up-regulated in the model group.Compared with the model group,Yiqi Huayu Jiedu Formula and the dismantling formula could significantly reduce the lung injury of ARDS model rats,and significantly down-regulate the expression of Rap1b,MAC-1 in alveolar lavage fluid and LFA-1 in lung homogenate.Conclusion The protective effect of Yiqi Huayu Jiedu Formula on LPS-induced ARDS model rats may be related to the reduction of Rap1b,LFA-1 and MAC-1 protein expression levels,and the reduction of neutrophil adhesion activity.