机械通气对急性肺损伤大鼠细胞凋亡的影响

目的 研究不同潮气量及不同呼气末正压(PEEP)水平对急性肺损伤(AU)大鼠支气管和肺组织细胞凋亡的影响,并初步探讨细胞凋亡在呼吸机相关性肺损伤(VILI)中的作用机制.方法 选用40只SD大鼠,制作ALI模型,随机(随机数字法)分为:(1)小潮气量组(LV组),潮气量8 mL/kg,不加PEEP;(2)大潮气量组,潮气量30 mL./kg,不加PEEP;(3)小潮气量+ 2PEEP组(LV2P组),潮气量8 mL/kg,同时给PEEP 2 cmH2O(1 crnH2O =0.098 kPa)；(4)小潮气量+5PEEP组(LV5P组),潮气量8 mL/kg,同时给PEEP 5 cmH2O；(5)小潮气量+8PEEP组(LV8P组),潮气量8 mL/kg,同时给PEEP 8 cmH2O.通气2h后处死动物,留取肺标本.用脱氧核糖核苷酸末端转移酶介导的末端标记法(TUNEL)分析肺组织中的细胞凋亡情况,用免疫组化法检测肺组织中caspase-3蛋白的表达及分布.结果 大潮气量组支气管和肺组织细胞凋亡的明显增加(P＜0.01),caspase-3蛋白酶表达最强.采用PEEP后,支气管和肺组织细胞凋亡减少,capase-3蛋白酶表达弱阳性,以LV5P组最为显著(P＜0.01).结论 小潮气量对肺组织有保护作用,采用PEEP后保护作用更加明显,细胞凋亡在VILI的发生中有重要作用.

Effects of mechanical ventilation on cell apoptosis in lung tissue of rat with acute lung injury

Objective To investigate the effects of different tidal volumes and positive end expiratory pressures on cell apoptosis in lung tissue of rats with acute lung injury.Methods Forty healthy male Sprague-Dawley rats were randomly(random number)divided into five groups,namely low tidal volume group(LV,VT 8 mL/kg),high tidal volume group(HV,VT 30 mL/kg),low tidal volume group with PEEP 2cmH2O(LV2P,VT 8 mL/kg,PEEP 2 cmH2O),low tidal volume group and PEEP 5cmH2O (LV5P,VT 8 mL/kg,PEEP 5 cmH2O)and low tidal volume group and PEEP 8 cmH2O(LV8P,VT 8 mL/kg,PEEP 8 cmH2O).After intravenous administration of oleic acid(OA,0.1 mL/kg),the rat model of acute lung injury was made.Mechanical ventilation was employed in rats of the experiment groups.Rats were sacrificed and their whole lungs were taken after mechanical ventilation for 2 hours.The transferase d-UTP end labeling assay(TUNEL)was used to define the extent and distribution of apoptotic cells in bronchus and lung tissues.The level of caspase-3 protein was determined by immunohistochemistry.Results The apoptotic cells on both alveolar septum and bronchial epithelium obviously increased with high level of caaspase-3 protein in HV group.The number of apoptotic cells obviously decreased with decrease in caspase-3 protein after PEEP ventilation.These changes were more significant in LV5P than those in other groups(P ＜ 0.01).Conclusions The mechanical ventilation with low tidal volume and PEEP produces protective effects on lung from injury.The cell apoptosis plays an important role in the course of VILI.