盐酸戊乙奎醚对大鼠内毒素性肺损伤时小窝蛋白-1表达的影响

Effect of penehyclidine hydrochloride on expression of caveolin-1 during LPS-induced lung injury in rats

目的评价盐酸戊乙奎醚对大鼠内毒素性肺损伤时小窝蛋白-1(Cav-1)表达的影响。方法SPF级健康雄性SD大鼠30只,体重170~190 g,采用随机数字表法分为3组(n=10):对照组(C组)、内毒素性肺损伤组(LI组)和盐酸戊乙奎醚组(PHCD组)。采用气管内注射LPS 0.2 ml(5 mg/kg)的方法制备内毒素性肺损伤模型。PHCD组于模型制备前1 h腹腔注射盐酸戊乙奎醚2 mg/kg(0.5 ml)。于模型制备后24 h时,取动脉血样,进行血气分析,采用ELISA法检测血清TNF-α和IL-1β浓度。随后处死大鼠,进行支气管肺泡灌洗,收集支气管肺泡灌洗液(BALF),计算PMNs百分比;取肺组织,观察病理学结果,采用比色法测定髓过氧化物酶(MPO)活性,计算湿重/干重比值(W/D比值);采用Western blot法测定Cav-1和核蛋白NF-κB的表达水平。结果与C组比较,LI组pH值和PaO2降低,PaCO2、BALF PMNs百分比、W/D比值和MPO活性升高,Cav-1表达下调,核蛋白NF-κB表达上调,血清TNF-α和IL-1β浓度升高(P<0.05)。与LI组比较,PHCD组pH值和PaO2升高,PaCO2、BALF PMNs百分比、肺组织W/D比值和MPO活性降低,Cav-1表达上调,核蛋白NF-κB表达下调,血清TNF-α和IL-1β浓度降低(P<0.05),肺组织病理学损伤减轻。结论盐酸戊乙奎醚减轻大鼠内毒素性肺损伤的机制可能与上调肺组织Cav-1表达,减轻炎症反应有关。

Objective To evaluate the effect of penehyclidine hydrochloride (PHC) on the expression of caveolin-1 (Cav-1) with lipopolysaccharide (LPS)-induced lung injury (LI)in rats. Methods Thirty SPF healthy male Sprague-Dawley rats, weighing 170-190 g, were divided into 3 groups (n=10 each) using a random number table method: control group (group C), LPS-induced LI group (group LI) and PHCD group.LI was produced by injecting LPS 0.2 ml (5 mg/kg) via the trachea in anesthetized rats. PHCD 0.5 ml (2 mg/kg) was intraperitoneally injected at 1 h before establishing the model in group PHCD. Arterial blood samples were collected at 24 h after establishing the model for blood gas analysis and for determination of serum tumor necrosis factor-alpha (TNF-α) and interleukin-1beta (IL-1β) concentrations by enzyme-linked immunosorbent assay. Rats were then sacrificed, and the lungs were removed. The main bronchus was lavaged, and the broncho-alveolar lavage fluid (BALF) was collected for calculation of the percentage of polymorphonuclear neutrophils (PMNs). Lung tissues were obtained for examination of pathological changes and for determination of myeloperoxidase(MPO)activity (by colorimetric assay), wet/dry weight ratio (W/D ratio), and expression of Cav-1 and nuclear factor kappa B (NF-κB) in nucleoprotein (by Western blot). Results Compared with group C, pH value and PaO2 were significantly decreased, the PaCO2, percentage of PMNs in BALF, W/D ratio and MPO activity were increased, the Cav-1 expression was down-regulated, the expression of NF-κB in nucleoprotein was up-regulated, and the serum TNF-α and IL-1β concentrations were increased in group LI (P<0.05). Compared with group LI, pH value and PaO2 were significantly increased, the PaCO2, percentage of PMNs in BALF, W/D ratio and MPO activity were decreased, the Cav-1 expression was up-regulated, the expression of NF-κB in nucleoprotein was down-regulated, and the serum TNF-α and IL-1β concentrations were decreased (P<0.05), and the pathological changes of lung tissues were significantly attenuated in group PHCD(P>0.05). Conclusion The mechanism by which PHC reduces LPS-induced LI may be related to up-regulating the expression of Cav-1 and mitigating inflammatory responses in lung tissues of rats.