普拉格雷对海水吸入型急性肺损伤的作用分析

doi:10.3877/cma.j.issn.1674-6902.2019.02.004

目的

探讨海水吸入型急性肺损伤中血小板与中性粒细胞的黏附情况，以及普拉格雷对其的干预作用。

方法

SD大鼠随机分为海水吸入1、2 h两个大组，组内随机分为空白对照组、海水处理组、溶剂处理组与普拉格雷处理组，每组8只大鼠。海水处理组在大鼠气管内灌注海水，溶剂处理组和普拉格雷处理组在海水处理之前4 h分别给予羟甲基纤维素钠溶液(CMC-Na)和普拉格雷灌胃处理。分别进行各组的病理切片HE染色，检测各组肺组织湿干比、血浆中炎症因子浓度，流式细胞术下检测血小板活化率以及与中性粒细胞的黏附情况，并进行肺泡灌洗液蛋白浓度测定以及髓过氧化物酶(MPO)活性测定。

结果

与正常对照组比较，气管内滴入海水后大鼠肺部组织出现明显炎症和水肿，HE染色显示其组织结构紊乱，肺泡间质充血水肿，并有大量炎症细胞浸润，肺泡间隔明显增厚，ELISA法测得全身炎症因子，如TNF-α、IL-1β以及TXB2的表达增高，血小板与中性粒细胞黏附率显著增加，肺泡灌洗液蛋白定量以及MPO活性增加，并且海水吸入2 h较1 h各项指标增高更为显著。普拉格雷干预可显著减轻海水导致的肺组织损伤，减轻肺组织结构紊乱与炎症细胞浸润，减少炎症因子TNF-α、IL-1β以及TXB2的释放，并且抑制血小板的活化以及与中性粒细胞的黏附。

结论

海水吸入型急性肺损伤中血小板与中性粒细胞的黏附率明显升高，普拉格雷可能通过降低此指标对肺损伤产生保护作用。

关键词: 肺损伤，海水, 普拉格雷, 血小板, 中性粒细胞

Objective

To observe the protective effect of Prasugrel on seawater aspiration-induced acute lung injury of SD rats and explore the possible mechanisms.

Methods

Seawater was instilled into the airway of SD rats. The rats were observed after 1 h or 2 h once acute lung injury was conducted. Then the rats were randomly divided into 3 experimental groups (n=8 in each group): seawater group, solvent sodium carboxmethylcellulose (CMC-Na) group and Prasugrel group. Other 8 healthy SD rats were taken as the normal control group. The rats were treated with CMC-Na solution or Prasugrel 4 h before seawater inhalation by intragastric administration in the CMC-Na group and Prasugrel group, respectively. Histological examination, measurements of wet to dry weight ratio of the lung samples, the expression of inflammatory cytokines and the protein concentration and myeloperoxidase (MPO) activity of the bronchoalveolar lavage fluid (BALF) were carried out after modeling. And the adhesion of the platelets with neutrophils was measured by flow cytometry.

Results

Compared with the normal control group, the inflammation and edema of the lung tissues were obvious in the three experimental groups. The sections of hematoxylin-eosin staining showed that the alveolar architecture distortion, pulmonary edema, interstitial thickening and infiltration of the inflammatory cells in the lung tissues were evident after seawater instillation. And the expression levels of TNF-α, IL-1β and TXB2 in the plasma measured by ELISA increased after seawater stimulation, as well as the protein concentration and MPO activity of BALF. The adhesion of platelets and neutrophils increased significantly. Meanwhile, the damage was more severe 2 h after seawater stimulation. However, pretreatment of Prasugrel attenuated the lung inflammation and edema, decreased the release of inflammatory factors, and inhibited the adhesion of platelets and neutrophils.

Conclusion

The adhesion of platelets and neutrophils after seawater aspiration-induced acute lung injury is increased notably. Prasugrel ameliorates the injury and reveals a remarkable protective effect by inhibiting the adhesion of platelets and neutrophils probably.

Key words: Acute lung injury, Seawater, Prasugrel, Platelets, Neutrophils

图1 不同处理组大鼠肺组织的病理切片，大鼠1 h与2 h肺组织的损伤；注：A：空白对照组；B：海水淹溺组；C：溶剂CMC-Na预处理组；D：普拉格雷预处理组(HE×20)

图2 不同处理组肺组织湿干比的比较，ELISA检测血浆TNF-α、IL-1β及TXB2水平；注：*表示P<0.05，**表示P<0.1，n.s.表示无统计学差异；A：空白对照组；B：海水淹溺组；C：溶剂CMC-Na预处理组；D：普拉格雷预处理组

图3 不同处理组肺泡灌洗液蛋白浓度及MPO活性的比较；注：*表示P<0.05；n.s.表示无统计学差异；A：空白对照组；B：海水淹溺组；C：溶剂CMC-Na预处理组；D：普拉格雷预处理组

图4 不同处理组血小板活化率以及血小板与中性粒细胞黏附率的比较；注：*表示P<0.05；n.s.表示无统计学差异；A：空白对照组；B：海水淹溺组；C：溶剂CMC-Na预处理组；D：普拉格雷预处理组

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Prasugrel ameliorates seawater aspiration-induced acute lung injury in rats by inhibiting adhesion of platelets and neutrophils

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Prasugrel ameliorates seawater aspiration-induced acute lung injury in rats by inhibiting adhesion of platelets and neutrophils