富氢液对脂多糖致小鼠急性肺损伤的影响

Effect of hydrogen-rich saline on lipopolysaccharide-induced acute lung injury in mice

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 increased, 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 decreased in group ALI＋ H2 （ P 〈 0.05）. Conclusion Hydrogen-rich saline can alleviate LPS-induced ALl via inhibition of in- flammatory response and apoptosis in mice.