目的: 研究甘草酸二铵脂质配体(DGLL)对大鼠急性肺损伤(ALI)及肺水肿的影响。方法: 雄性SD大鼠123只,体重170～200 g,(6±1)周龄,灌胃DGLL(30、60、120 mg/kg),1 h后通过腹腔内注射脂多糖(LPS)10 mg/kg建立大鼠ALI模型。6 h后,使用HE染色技术评估肺损伤。使用肺湿干重比、支气管肺泡灌洗液(BALF)中的蛋白质含量和肺组织中的Evans蓝(EB)来评估肺水肿。用ELISA法测定肺组织中肿瘤坏死因子(TNF-α)和白细胞介素(IL-1β)的表达水平。通过免疫组织化学染色法检测髓过氧化物酶(MPO)的表达水平。Western blotting法测定细胞间黏附分子(ICAM-1)、血管内皮钙黏蛋白(VE-cadherin)、黏附蛋白(ZO-1)、连接黏附分子(JAM-1)等与肺部炎症和微血管通透性有关的蛋白表达水平。结果: MPO免疫反应性降低,大鼠肺组织中TNF-α、IL-1β和ICAM-1的表达水平下降,证明DGLL缓解了LPS诱导的ALI。此外,DGLL可以抑制LPS诱导的肺水肿,降低BALF的蛋白浓度及EB外渗。DGLL也降低了VE-cadherin的表达水平以及抑制LPS引起的肺组织中的连接蛋白,包括ZO-1、JAM-1的表达。结论: DGLL对LPS诱导的大鼠ALI表现出保护作用,同时也抑制了炎症细胞的浸润和微血管屏障的破坏。
Objective: To study the effects of diammonium glycyrrhizinate (DGLL) on acute lung injury (ALI) and pulmonary edema in rats. Methods: A total of 123 male SD rats, weighing 170~200 g, (6±1) weeks old, were intragastrically administrated with DGLL (30, 60, 120 mg/kg). After 1 h, the ALI model was established by intraperitoneal injection of LPS (10 mg/kg ). After 6 h, the lung injury was evaluated by HE staining. Lung wet to dry weight ratio, protein content in bronchoalveolar lavage fluid (BALF) and Evans blue (EB) in lung tissue were used to evaluate pulmonary edema. The expression levels of tumor necrosis factor (TNF-α) and interleukin (IL-1β) in lung tissue were determined by ELISA. The expression level of myeloperoxidase (MPO) was detected by immunohistochemical staining. The expression levels of intercellular adhesion molecule (ICAM-1), vascular endothelial cadherin (VE-cadherin), adhesion protein (ZO-1), and connective adhesion molecule (JAM-1) related to lung inflammation and microvascular permeability were determined by Western blotting. Results: The immunoreactivity of MPO was decreased, and the expression levels of TNF-α, IL-1β and ICAM-1 in lung tissue of rats were decreased, indicating that DGLL alleviated LPS-induced ALI. In addition, DGLL could inhibit LPS-induced pulmonary edema, reduce BALF protein concentration and EB extravasation. DGLL also could reduce the expression level of VE-cadherin in endothelial cells and inhibited the expression of connexins in lung tissue induced by LPS, including ZO-1 and JAM-1. Conclusion: DGLL has a protective effect on LPS-induced ALI in rats, and also inhibits the infiltration of inflammatory cells and the destruction of microvascular barrier.
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