摘要
目的:分析丹参酮IIA(tanshinone IIA, TIIA)与苦参碱(matrine, MAT)组合用药对脂多糖(lipopolysaccharide, LPS)致小鼠急性肺损伤(acute lung injury, ALI)的预防及治疗作用，探讨作用机制。方法:选择60只雄性C57小鼠，随机分为五组：对照组，LPS组，TIIA+LPS组(TIIA组)，MAT+LPS组(MAT组)，TIIA+MAT+LPS组(TIIA+MAT组)，每组12只。TIIA组给予TIIA 10 mg/kg剂量灌胃，MAT组给予MAT 40 mg/kg灌胃，TIIA+MAT组给予TIIA 10 mg/kg+MAT 40 mg/kg灌胃，灌胃3 d，对照组及LPS组用同等体积的无菌生理盐水灌胃处理相同时间。末次给药1 h后除对照组外每组给予LPS 5 mg/kg腹腔注射行急性肺损伤造模，对照组注射同等体积的无菌生理盐水。造模6 h后，取肺组织，HE染色观察肺组织形态学改变，测定湿/干重比值(W/D)，ELISA法测定肺泡灌洗液中肿瘤坏死因子-α(TNF-α)、白介素-1(IL-1)、白介素-6(IL-6)，试剂盒法测定肺组织匀浆中髓过氧化物酶(MPO)活力及肺泡灌洗液(BALF)中蛋白含量。结果:形态学观察表明LPS组肺组织明显充血、水肿，有大量炎性细胞浸润，在TIIA组、MAT组、TIIA+MAT组内毒素致肺损伤减轻，TIIA+MAT组表现明显，肺W/D比值降低(P<0.05)。单独应用TIIA或MAT时肺泡灌洗液中IL-1、IL-6、TNF-α有不同程度下降(P>0.05)，TIIA+MAT组肺泡灌洗液中TNF-α、IL-1、IL-6较LPS相应时相点下降(P<0.05)。BALF中蛋白浓度LPS组较对照组升高，应用TIIA或MAT后有不同程度下降(P<0.05)，TIIA+MAT组中蛋白浓度较LPS组降低(P<0.01)。肺组织匀浆MPO活力LPS组较对照组升高，单独应用TIIA或MAT有轻度下降，无统计学意义(P>0.05)，TIIA+MAT组较LPS组MPO活力降低(P<0.05)。结论:丹参酮IIA及苦参碱能减轻LPS致小鼠急性肺损伤，两者配合使用效果较单药使用效果佳。
Abstract
Objective:To observe the preventive and therapeutic effects of tanshinone IIA (TIIA) and matrine (MAT) in the prevention and treatment of lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice, and to preliminarily explore its mechanism.Methods:Sixty male C57 mice were randomly divided into five groups: control group, LPS group, TIIA+ LPS group (TIIA group), MAT+ LPS group (MAT group), TIIA+ MAT+ LPS group (TIIA+ MAT group), with 12 animals in each group. Among them, TIIA group was given TIIA 10 mg/kg dose of gavage, MAT group was given MAT 40 mg/kg gavage, TIIA + MAT group was given TIIA 10 mg/kg + MAT 40 mg/kg gavage for a total of three days, and the control group and LPS group were treated with the same volume of sterile normal saline for the same time. 1 h after the last dose, except for the control group, all groups were given LPS 5 mg/kg intraperitoneal injection for acute lung injury modeling, and the control group was injected with the same volume of sterile normal saline. After 6 hours of molding, lung tissue was taken and HE staining was used to observe the morphological changes of lung tissue, the wet/dry weight ratio (W/D) was determined, tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1), interleukin-6 (IL-6) in bronchoalveolar lavage fluid was determined by ELISA method, and myeloperoxidase (MPO) activity and protein content in bronchoalveolar lavage fluid(BALF) in lung tissue homogenate was determined by kit method.Results:Morphological observations showed that the lung tissue in the LPS group was significantly hyperemic and edematous, and there was a large number of inflammatory cell infiltrates, while the lung damage caused by endotoxin in the TIIA, MAT group and TIIA+ MAT group was reduced, especially in the TIIA+ MAT group, and the lung W/D ratio was reduced (P<0.05). When TIIA or MAT was applied alone, IL-1, IL-6 and TNF-α in alveolar lavage solution decreased to varying degrees, but there was no significant significance (P>0.05), and the concentrations of TNF-α, IL-1 and IL-6 in alveolar lavage solution in the TIIA+ MAT group decreased significantly compared with the corresponding phase points of LPS (P<0.05). The protein concentration in the LPS group in BALF was significantly higher than that in the control group, and decreased to varying degrees after TIIA or MAT (P<0.05), but the protein concentration in the TIIA+ MAT group was significantly lower than that in the LPS group (P<0.01). The MPO viability LPS group of lung tissue homogenate was significantly higher than that of the control group, and there was a slight decrease in TIIA or MAT alone, but there was no significant (P>0.05), while the TIIA+ MAT group was significantly lower than that in the LPS group. Statistically significant (P<0.05).Conclusion:Tanshinone IIA and matrine can reduce acute lung injury caused by LPS, but the combination effect of the two is better than that of single agent.
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