NOX2抑制剂gp91 ds-tat缓解瓦斯爆炸引起的大鼠肺损伤

NOX2 inhibitor gp91 ds-tat alleviated lung injury induced by gas explosion in rats

目的研究还原型烟酰胺腺嘌呤二核苷酸磷酸氧化酶(nicotinamide adenine dinucleotide phosphate oxidase,NOX2)抑制剂gp91 ds-tat对瓦斯爆炸肺损伤大鼠的影响。方法90只SD雄性大鼠随机分为空白对照组、瓦斯爆炸肺损伤模型组、gp91 ds-tat治疗组,每组30只。将模型组和治疗组大鼠放置于巷道距离起爆点240 m处。爆炸后立即一次性通过腹腔注射给予治疗组大鼠gp91 ds-tat(1 mg/kg)。3组大鼠分别于治疗后24 h、72 h、7 d(每组每次10只)麻醉后腹主动脉取血并处死,取肺组织包埋用于制作切片样品;冷冻肺组织用于三磷酸腺苷(ATP)、α-酮戊二酸脱氢酶(α-KGDH)、线粒体异柠檬酸脱氢酶(ICDHm)、磷酸果糖激酶(PFK)检测;采用蛋白免疫印迹法、实时荧光定量PCR检测大鼠肺组织NOX2、过氧化氢酶(CAT)及线粒体功能相关蛋白基因的表达。结果与对照组比较,模型组大鼠出现肺脏出血、塌陷,CAT、AMP依赖的蛋白激酶(AMPK)、线粒体关键转录分子A(TFAM)表达降低(P<0.05),而NOX2表达升高(P<0.05)。与模型组相比,gp91 ds-tat治疗组大鼠的α-KGDH、ICDHm、PFK的活力及ATP含量升高(P<0.05),CAT、TFAM、mRNA相对表达水平提高(P<0.05),而NOX2的表达降低(P<0.05)。结论gp91 ds-tat可能通过调控肺内的能量代谢紊乱和氧化应激状态减轻瓦斯爆炸所致大鼠肺损伤的程度,其作用可能与抑制NOX2的表达相关。

Objective To assess the effects of NOX2(nicotinamide adenine dinucleotide phosphate oxidase)inhibitor gp91ds-tat on lung injury of rats caused by gas explosion.Methods A total of 90 SD male rats were randomly divided into control group,gas explosion lung injury model group,and NOX2 inhibitor gp91 ds-tat group(n=30/group).The rats in the model group and gp91 ds-tat group were placed in the alleyway 240 m away from the point of detonation.After the explosion,the rats of the gp91 ds-tat group were immediately given once intraperitoneal injection of gp91 ds-tat(1 mg/kg).Then the rats were anaesthetized with blood taken from the abdominal aorta at 24 h,72 h and 7 d(10 rats in each group)and then executed,respectively.ATP,α-KGDH,ICDHm and PFK were detected in frozen lung tissues.The expression levels of NOX2,CAT protein and mRNA as well as the expression of mitochondrial function-related protein genes were also detected with western blotting and real-time quantitative PCR in rat lung tissues.Results Lung bleeding and collapse were observed in the model group,and the expression of CAT,AMPK,TFAM decreased(P<0.05),but the expression of NOX2 increased significantly(P<0.05).Compared with the model group,levels ofα-KGDH,ICDHm,PFK and ATP in gp91 ds-tat treatment group increased(P<0.05),the relative mRNA expression of CAT and TFAM increased(P<0.05),and the expression of NOX2 decreased(P<0.05).Conclusions Gp91 ds-tat could reduce the lung injury degree in gas explosion injury by regulating the disorder of energy metabolism and oxidative stress status in the lung,and its effect may be related to the inhibition of NOX2 expression.