"丹蛇"组方对小鼠急性肺损伤的治疗作用

doi:10.3877/cma.j.issn.1674-6902.2024.02.001

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。

关键词: 急性肺损伤, 丹参酮ⅡA, 蛇床子素, 脂多糖

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, Tanshinone ⅡA group, Osthole group and Tanshinone ⅡA+ 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 Tanshinone ⅡA group and Osthole group was 50%, and the 72 h survival rate of mice in Tanshinone ⅡA+ 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 Tanshinone ⅡA group and Osthole group was less than that in LPS group, and the repair of pathological structure of lung tissue in Tanshinone ⅡA+ Osthole group was more obvious. W/D results showed that compared with LPS group, the pulmonary edema of Tanshinone ⅡA group and Osthole group were not improved, and the pulmonary edema of Tanshinone ⅡA+ 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 Tanshinone ⅡA group and Osthole group, and significantly decreased in Tanshinone ⅡA+ 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 Tanshinone ⅡA group and Osthole group were decreased compared with LPS group(P<0.05). The secretion of pulmonary inflammatory factor decreased significantly in Tanshinone ⅡA+ Osthole group (P<0.05).

Conclusion

Tanshinone ⅡA+ 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. Tanshinone ⅡA+ Osthole formula has therapeutic effect on LPS-induced acute lung injury in mice.

Key words: Acute lung injury, Tanshinone ⅡA, Osthole, Lipopolysaccharide

图1 小鼠的存活率。对照组vs. LPS组P<0.05；LPS组vs.丹参酮ⅡA组，P<0.05；LPS组vs.蛇床子素组，P<0.05；LPS组vs.丹参酮ⅡA＋蛇床子素组，P<0.05

图2 小鼠肺部形态学变化。注：A：对照组；B：丹参酮ⅡA＋蛇床子素组；C：LPS组；D：丹参酮ⅡA组、E蛇床子素组

图3 小鼠的肺湿重/干重比(W/D)。注：Control：对照组；LPS：脂多糖

图4 小鼠BALF中蛋白含量和细胞计数变化。注：Control：对照组；LPS：脂多糖

图5 小鼠组织匀浆中MPO活力变化。注：Control：对照组；LPS：脂多糖

图6 小鼠BALF中炎性因子变化

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Therapeutic effect of "Tanshinone ⅡA+ Osthole" formula on acute lung injury in mice

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Therapeutic effect of "Tanshinone ⅡA+ Osthole" formula on acute lung injury in mice