盐酸戊乙奎醚对ARDS大鼠炎性因子的影响

Anti-inflammatory effects of penehyclidine hydrochloride on acute respiratory distress syndrome in rats

目的 探讨盐酸戊乙奎醚对油酸致急性呼吸窘迫综合征（ARDS）大鼠的抗炎作用.方法 按随机数字表法将30只成年雄性SD大鼠均分为对照组、模型组及治疗组.各组大鼠均经股静脉置管,对照组注入生理盐水1.1 mL/kg;模型组注射0.1 mL/kg油酸后随即注入生理盐水1.0 mL/kg;治疗组注入0.1 mL/kg油酸后随即注入1.0 mL/kg盐酸戊乙奎醚.制模后3 h,对各组大鼠进行血气分析,计算氧合指数（PaO2/FiO2）,用酶联免疫吸附试验（ELISA）测定血清白细胞介素（IL-1、IL-6）含量;处死大鼠取肺组织,计算肺湿/干重（W/D）比值,镜下观察肺组织形态学改变;用比色法测定超氧化物歧化酶（SOD）活性、丙二醛（MDA）含量;免疫组化法检测肺组织肿瘤坏死因子-α（TNF-α）表达.Pearson相关分析肺W/D比值与各指标的相关性.结果 与对照组比较,模型组大鼠PaO2/FiO2明显下降,肺W/D比值明显升高,血清IL-1、IL-6水平及肺组织MDA含量显著升高,肺组织SOD活性明显下降;光镜下可见肺泡腔内大量渗出液、中性粒细胞和红细胞,肺泡间隔明显增宽、水肿;电镜下可见Ⅱ型肺泡上皮细胞板层小体变性、局部破坏及排空现象明显;免疫组化显示,肺组织TNF-α阳性表达明显增多.与模型组比较,治疗组PaO2/FiO2明显改善〔mmHg（1 mmHg=0.133 kPa）：204.42±31.61比113.91±47.78,P〈0.05〕,肺W/D比值明显降低（5.80±0.44比6.82±0.59,P〈0.01）,血清IL-1、IL-6水平及肺组织MDA含量明显下降〔IL-1（μg/L）：18.38±0.28比20.04±0.39,IL-6（μg/L）：12.64±0.67比14.28±1.33,MDA（nmol/mg）：3.95±0.28比5.17±0.29,均P〈0.05〕,肺组织SOD活性明显升高（U/mg：48.75±2.41比45.09±1.69,P〈0.01）,组织形态学及病理学改变明显减轻,肺组织中TNF-α阳性表达明显减少〔阳性细胞率：（25.80±3.44）％比（38.82±3.59）％,P〈0.01〕.结论 盐酸戊乙奎醚通过降低肺W/D比值,抑制氧化应激和炎症反应来有效改善油酸致ARDS大鼠的氧合、肺损伤及肺水肿程度.

Objective To investigate the anti-inflammatory effect of penehyclidine hydrochloride on oleic acid-induced acute respiratory distress syndrome （ARDS） in rats.Methods According to randomize number table method, 30 adult male Sprague-Dawley （SD） rats were divided into control group, model group and treatment group. Catheters were placed in femoral vein in each group. The control group was injected with 1.1 mL/kg physiological saline; the model group was injected with 0.1 mL/kg oleic acid and then injected with 1.0 mL/kg normal saline to establish ARDS model; the treatment group was injected with 0.1 mL/kg oleic acid and then injected with 1.0 mL/kg penehyclidine hydrochloride. At 3 hours after the model was established, blood gas analysis was carried out in each group, oxygenation index （PaO2/FiO2） was calculated, and the levels of serum interleukins （IL-1, IL-6） were measured by enzyme linked immunosorbent assay （ELISA）. Rats were sacrificed to harvest lung tissue, and the lung wet/dry ratio （W/D） was calculated; the morphological changes of lung tissue was observed under microscope; the superoxide dismutase （SOD） activity and malondialdehyde （MDA） levels were detected by colorimetry; the expression of tumor necrosis factor-α（TNF-α） in lung tissue was detected by immunohistochemical method. The correlations between lung W/D ratio and various indicators were analyzed using Pearson correlation test.Results Compared with the control group, PaO2/FiO2 in model group was significantly decreased, lung W/D ratio was significantly increased, serum IL-1, IL-6 levels and lung tissue MDA content were significantly increased, lung tissue SOD activity was significantly decreased; the alveolar space was filled with exudate, neutrophils and red blood cells, and there were obvious edema and broadening in pulmonary interstitial and alveolar under light microscope; the type Ⅱ alveolar epithelial cells were partly destroyed, accompanied by lamellar body degeneration and emptying phenomenon under electron microscope; and immunohistochemistry showed that the positive expression of TNF-α in lung tissue was significantly increased. Compared with the model group, PaO2/FiO2 in the treatment group was significantly improved [mmHg （1 mmHg = 0.133 kPa）： 204.42±31.61 vs. 113.91±47.78,P 〈 0.05], the lung W/D ratio was significantly decreased （5.80±0.44 vs. 6.82±0.59,P 〈 0.01）, serum IL-1, IL-6 levels and lung tissue MDA content were significantly decreased [IL-1 （μg/L）： 18.38±0.28 vs. 20.04±0.39, IL-6 （μg/L）： 12.64±0.67 vs. 14.28±1.33, MDA （nmol/mg）： 3.95±0.28 vs. 5.17±0.29, allP 〈 0.05], the activity of SOD in lung tissue was significantly increased （U/mg： 48.75±2.41 vs. 45.09±1.69,P 〈 0.01）, histological and pathological changes were significantly reduced, and the positive expression of TNF-α in lung tissue was significantly reduced [positive cell rate： （25.80±3.44）% vs. （38.82±3.59）%,P 〈 0.01].Conclusion Penehyclidine hydrochloride can effectively improve the oxygenation, alleviate lung injury and reduce pulmonary edema in oleic acid induced ARDS rat by decreasing lung W/D ratio, inhibiting oxidative stress and inflammatory response.