目的评价富氢液对脂多糖(LPS)致小鼠急性肺损伤的影响。方法成年雄性C57BL/6小鼠32只,体重20-25g,采用随机数字表法,将其分为4组(n=8):对照组(c组)、富氢液组(H2组)、急性肺损伤组(ALI组)和急性肺损伤+富氢液组(Au+H2组)。Au组和ALI+H2组分别雾化吸人LPS25μg(溶于PBS中)制备Au模型,c组和H2组分别雾化吸入无菌PBS50μl。H2组和ALI+H2组雾化吸入PBS或LPS后1和12h时腹腔注射0.6mmol/L富氢液5ml/kg。LPS或PBS处理后24h时,机械通气15min,行动脉血气分析,计算氧合指数。然后收集支气管肺泡灌洗液(BALF),测定蛋白浓度,计数中性粒细胞(PMN)。采用ELISA法检测BALF中TNF-α、IL-1β、IL-6和高迁移率族蛋白1(HMGB1)的浓度。然后处死小鼠,取肺组织,行病理学损伤评分,测定湿重,干重(W/D)比、髓过氧化物酶(MPO)和caspase-3的恬性,计算细胞凋亡指数(AI)。结果与C组比较,H2组氧合指数、BALF总蛋白、TNF-α、IL-1β、IL-6和HMGB1的浓度和PMN计数、肺组织病理学损伤评分、W/D比、MPO和caspase-3的活性以及AI差异均无统计学意义(P〉0.05),Au组和ALI+H2组氧合指数降低,BALF蛋白、TNF-α、IL-1β、IL-6和HMGB1的浓度、PMN计数、肺组织病理学损伤评分、W/D比、MPO和caspase-3的活性以及AI均升高(P〈0.05);与AL1组比较,Au+H2组氧合指数升高,BALF总蛋白、TNF-α、IL-1β、IL-6和HMGB1的浓度、PMN计数、肺组织病理学损伤评分、W/D比、MPO和caspase-3的活性以及AI均降低(P〈0.05)。结论富氢液可减轻LPS致小鼠急性肺损伤,可能与其抗炎和抗凋亡作用有关。
Objective To investigate the effects of hydrogen-rich saline on lipopolysaccharide (LPS)-in- duced acute lung injury (ALI) in mice .Methods Thirty-two male C57BL/6 mice, weighing 20-25 g, were ran- domly divided into 4 groups ( n = 8 each) : control group (group C), hydrogen-rich saline group (group H2 ), Aid group and ALI + hydrogen-rich saline group (group ALI+ H2 ). ALI was induced by inhaled aerosolized LPS 25 μg (dissolved in PBS) in groups ALI and ALI+ H2 . Aerosolized PBS 50 μl was inhaled in groups C and H2 .0.6 mmol/L hydrogen-rich saline 5 ml/kg was injected intraperitoneally at 1 and 12 h after aerosolized PBS or LPS was inhaled in groups H2 and Aid + H2, respectively. At 24 h after PBS or LPS treatment, the animals were mechani- cally ventilation for 15 min, and arterial blood gas analysis was performed. The oxygenation index (PaO2/FiO2 ) was calculated. Bronchoalveolar lavage fluid (BALF) was collected for determination of total protein concentration and polymorphonuclear neutrophils (PMN) count. The concentrations of TNF-α,IL-1β,IL-6and high mobility group box-1 (HMGB-1 ) protein in BALF were determined by ELISA. Then the animals were sacrificed and thelungs were obtained for determination of W/D lung weight ratio (W/D ratio), and myeloperoxidase (MPO) and caspase-3 activities. The pathological changes of the lung were scored. Apoptosis index (AI) was calculated. Re- suits Compared with group C, no significant difference was found in PaO2/FiO2, the concentrations of protein, TNF-α,IL-1β,IL-6 and HMGB1, and PMN count in BALF, lung injury score, W/D ratio, MPO and caspase-3 activities and AI in group H2 (P 〉 0.05), and PaO2/FiO2 was significantly decreased, the concentrations of total protein, TNF-α,IL-1β,IL-6 and HMGB1, and PMN count in BALF, lung injury score, W/D ratio , MPO and caspase-3 activities, and AI were increased in groups ALI and ALI+ H2 (P 〈 0.05). Compared with group ALl, PaO2/FiO2 was significantly