呼吸机相关性肺损伤中胱天蛋白酶1／3介导的细胞凋亡机制及乌司他丁的干预作用

摘要

Objective To explore the mechanism of caspase1/3-mediated apoptosis involved in lung injury and the intervention of ulinastatin.Methods A total of 36 male SD rats were divided into 3 groups randomly:the control group,model group and ulinastatin group (n=12).Rats in the control group did not receive mechanical ventilation;rats in model group and ulinastatin group were given H-VT for modelling;rats in ulinastatin group were given intravenous pre-infusion of ulinastatin(105 U/kg);rats in model group were given same dosage of 0.9%sodium chloride solution.The biochemical indictors including tumor necrosis fac-tor α( TNF-α) , malondialdehyde ( MDA ) , superoxide dismutase ( SOD ) , and glutathione peroxidase ( GSH-PX) in the bronchoalveolar lavage fluids of the rats in three groups were assayed by biochemical reactions . The expression of caspase 1/3,p53 and BAX/Bcl-2 were detected by western blotting.Results In the mod-el group,levels of SOD and GSH-PX in the model group were significantly decreased ,while levels of MDA and TNF-αwere significantly increased;the expressions of caspase 1/3,p53 and BAX were up-regulated, while expression of Bc1-2 were down-regulated.The expressions of aforementioned proteins in ulinastatin group were improved partially (relative optical densities of BAX, Bcl2, P53 and caspase 1/3 proteins were as follows:0.25∶0.81∶0.67,1.18∶0.24∶0.61,0.49∶1.28∶0.68,0.38∶0.61∶0.69,0.42∶1.22∶0.74). Conclusion Ulinastatin can alleviate ventilator related lung injury caused by apoptosis via suppressing the expression of caspase 1/3 and p53 .%目的：探讨呼吸机相关肺损伤中胱天蛋白酶（caspase）1／3介导的细胞凋亡机制及乌司他丁的干预作用。方法将36只大鼠完全随机分为3组：对照组、模型组及乌司他丁组，各12只。对照组大鼠未进行机械通气，模型组和乌司他丁组大鼠给予大潮气量造模，乌司他丁组大鼠在通气前给予静脉预灌注乌司他丁（10万U／kg），模型组大鼠给予等剂量0.9％氯化钠溶液。分析各组大鼠支气管肺泡灌洗液血清生化指标肿瘤坏死因子α（ TNF-α）、丙二醛、超氧化物歧化酶（ SOD）及谷胱甘肽过氧化物酶（ GSH-PX）及细胞凋亡相关因子caspase-1／3、p53、B淋巴细胞瘤基因2／Bcl-2相关X蛋白（BAX／Bcl-2）的表达变化。结果模型组灌洗液 TNF-α、丙二醛水平显著高于对照组[（490±51） mg／L 比（157±16） mg／L，（62±20） mg／L比（32±11） mg／L，P＜0.05]，而SOD及GSH-PX水平显著低于对照组[（226±41） kU／L 比（401±61） kU／L，（267±33） kU／L 比（62.4±52） kU／L， P＜0.05]，调亡相关蛋白caspase-1／3及p53、BAX显著高于对照组，Bcl-2显著低于对照组。与模型组比较，乌司他丁组上述异常表达蛋白得到部分改善，但仍未恢复到对照组水平（ BAX／Bcl-2／p53／caspase-1／3相对光密度＝0.25∶0.81∶0.67／1.18∶0.24∶0.61／0.49∶1.28∶0.68／0.38∶0.61∶0.49／0.42∶1.22∶0.74）。结论乌司他丁抑制 caspasse-1／3及 p53通路改善细胞凋亡引起的呼吸机性肺损伤。