“丹蛇”组方对小鼠急性肺损伤的治疗作用

Therapeutic effect of“TanshinoneⅡA+Osthole”formula on acute lung injury in mice

目的分析中药组方-丹参酮ⅡA+蛇床子素对脂多糖(lipopolysaccharide,LPS)致小鼠急性肺损伤(acute lung injury,ALI)的治疗作用。方法选择50只C57小鼠,随机分为对照组(生理盐水)、LPS组、丹参酮ⅡA组、蛇床子素组、丹参酮ⅡA+蛇床子素组,每组10只,气管内滴注LPS建立小鼠急性肺损伤模型,造模24 h后观察小鼠存活率、肺组织形态学变化,检测72 h左肺湿/干重比值(W/D)、支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中总细胞计数,采用ELISA法检测总蛋白、髓过氧化物酶(myeloperoxidase,MPO)和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)、白细胞介素-1(interleukin-1,IL-1)。结果LPS组小鼠24 h存活率为0,丹参酮ⅡA组、蛇床子素组72 h存活率为50%,丹参酮ⅡA+蛇床子素组72 h存活率约75%,存活率提高;HE染色结果显示,丹参酮ⅡA组、蛇床子素组较LPS组肺部组织病理结构破坏减轻,丹参酮ⅡA+蛇床子素组肺部组织病理结构修复情况较明显;W/D结果显示,丹参酮ⅡA组、蛇床子素组较LPS组肺部水肿无改善、丹参酮ⅡA+蛇床子素组肺部水肿情况减轻(P<0.05)。BALF中总蛋白含量和细胞计数显示,与LPS组相比,丹参酮ⅡA组、蛇床子素组肺部蛋白渗出和炎细胞分泌减轻,丹参酮ⅡA+蛇床子素组明显减轻(P<0.05)。MPO活力结果与BALF中细胞计数结果类似;丹参酮ⅡA组、蛇床子素组较LPS组肺部炎性因子TNF-α、IL-6、IL-1分泌减少(P<0.05)。丹参酮ⅡA+蛇床子素组肺部炎性因子分泌减少明显(P<0.05)。结论丹参酮ⅡA+蛇床子素组方能减少对LPS致小鼠肺部炎症因子分泌,减轻炎性细胞浸润、肺泡蛋白渗出、炎细胞漏出和肺部水肿,改善肺部病理组织结构,提高小鼠存活率。丹参酮ⅡA+蛇床子素组方对LPS致小鼠急性肺损伤具有治疗作用。

Objective To analyze the therapeutic effects of Tanshinone(ⅡA)+Ossathol in the treatment of lipopolysaccharide(LPS)-induced acute lung injury(ALI)in mice.Method A total of 50 C57 mice were randomly divided into control group(normal saline),LPS group,TanshinoneⅡA group,Osthole group and TanshinoneⅡA+Osyhole group,with 10 mice in each group.The mice acute lung injury model was established by endotracheal infusion of LPS.The survival rate and lung histopathology of mice were observed after 24 hours of the model was made.The left lung wet/dry weight ratio(W/D)and the total cell count in bronchoalveolar lavage fluid(BALF)at 72 h were detected.Total protein,myeloperoxidase(MPO),tumor necrosis factor-α(TNF-α)and interleukin-6(interleukin-6)and interleukin-1(IL-1)were detected by ELISA.Results The 24 h survival rate of mice in LPS group was 0,the 72 h survival rate of mice in TanshinoneⅡA group and Osthole group was 50%,and the 72 h survival rate of mice in TanshinoneⅡA+Osthole group was about 75%,and the survival rate was improved.The results of HE staining showed that the damage of pathological structure of lung tissue in TanshinoneⅡA group and Osthole group was less than that in LPS group,and the repair of pathological structure of lung tissue in TanshinoneⅡA+Osthole group was more obvious.W/D results showed that compared with LPS group,the pulmonary edema of TanshinoneⅡA group and Osthole group were not improved,and the pulmonary edema of TanshinoneⅡA+Osthole group was alleviated(P<0.05).The total protein content and cell count in BALF showed that compared with LPS group,the pulmonary protein exudation and inflammatory cell secretion were reduced in TanshinoneⅡA group and Osthole group,and significantly decreased in TanshinoneⅡA+Osthole group(P<0.05).The results of MPO activity were similar to those of cell count in BALF.The secretion of pulmonary inflammatory factors TNF-α,IL-6 and IL-1 in TanshinoneⅡA group and Osthole group were decreased compared with LPS group(P<0.05).The secretion of pulmonary inflammatory factor decreased significantly in TanshinoneⅡA+Osthole group(P<0.05).Conclusion TanshinoneⅡA+Osthole formula can reduce the secretion of lung inflammatory factors induced by LPS,relieve inflammatory cell infiltration,alveolar protein exudation,inflammatory cell leakage and pulmonary edema,improve lung pathological structure and increase the survival rate of mice.TanshinoneⅡA+Osthole formula has therapeutic effect on LPS-induced acute lung injury in mice.