全氟化碳对大鼠肺缺血再灌注损伤保护作用的实验研究
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摘要
目的:通过腹腔注射全氟化碳(C8F18)对大鼠肺缺血再灌注损伤(lung ischemical reperfusion injury,LIRI)起到保护作用。方法:将健康清洁SD大鼠随机分空白组、对照组、实验组。建立大鼠LIRI模型。空白组(NC组)开胸不进行LIRI处理。对照组(NS组)术前48 h腹腔注射0.9%NS后行LIRI处理;实验组(PFC组)术前48 h腹腔注射C8F18后行LIRI处理。检测3组实验动物动脉血气分析、肿瘤坏死因子(TNF-α)、肺组织水通道蛋白(AQP1)及钠通道蛋白(ENaC)的表达。结果:NS组与PFC组2 h时间点比较PaCO2值差异有统计学意义(P <0.05),4 h、6 h时间点两组PaCO2值差异无统计学意义(P> 0.05)。PFC组与NS组各时间点比较全血TNF-α含量下降(P <0.05)。AQP1蛋白的表达以空白组最明显,NS组AQP1蛋白染色明显减少,以2h组最明显,各时间点与空白组比较均下降(P <0.05)。PFC组AQP1蛋白较NS组有显著增加(P <0.05)。ENaC蛋白表达以NC组最明显,NS组ENaC蛋白染色明显减少,各时间点与NC组比较均显著下降(P <0.05),PFC组ENaC蛋白较NS组无差异(P>0.05)。结论:全氟化碳大鼠腹腔注射后,在大鼠LIRI中动脉血Pa02增加,PaCO2下降,促进肺气体交换。使全血中TNF-α浓度下降,从而起到保护肺组织的作用。同时能增加肺组织AQP1、ENaC蛋白的表达,有效减轻肺水肿及炎症反应。
Objective: To investigate the protective effect of intraperitoneal injection of perfluorocarbon(C8 F18)on lung ischemia-reperfusion injury(LIRI) in rats. Methods: A total of 90 healthy and clean Sprague-Dawley rats were randomly divided into blank group, control group, and experimental group. A LIRI model was established in rats. The blank group(NC group) underwent thoracotomy without LIRI treatment. The control group(NS group) received intraperitoneal injection of 0.9% NS at 48 h before thoracotomy and LIRI treatment. The experimental group(PFC group) received intraperitoneal injection of C8 F18 at 48 h before thoracotomy and LIRI treatment. Arterial blood gas parameters and the expression of tumor necrosis factor(TNF-α), lung aquaporin(AQP1), and sodium channel protein(ENaC) were measured in the three groups. Results: There was a significant difference in partial pressure of carbon dioxide(PaCO2) at 2 h between the NS group and the PFC group(P < 0.05), but there were no significant differences in PaCO2 at 4 and 6 h between the two groups(P > 0.05). The PFC group had a significantly lower level of tumor necrosis factor-alpha(TNF-α) in whole blood at each time point compared with the NS group(P <0.05). Among the three groups, the NC group had the highest expression of AQP1. The staining of AQP1 in the NS group was significantly reduced, especially at 2 h, and the NS group had significantly lower expression of AQP1 at each time point compared with the NC group(P < 0.05). The PFC group had significant higher expression of AQP1 compared with the NS group(P < 0.05). Among the three groups, the NC group had the highest expression of ENaC.The staining of ENaC in the NS group was significantly reduced, and the NS group had significantly lower expression of ENaC at each time point compared with the NC group(P < 0.05). There was no significant difference in the expression of ENaC between the PFC group and the NS group(P > 0.05). Conclusion: In the LIRI rat model, intraperitoneal injection of perfluorocarbon has a protective effect on the lung tissue by promoting pulmonary gas exchange — increasing PaO2 in arterial blood and decreasing PaCO2 and by reducing the level of TNF-α in whole blood. At the same time, it can effectively alleviate pulmonary edema and inflammatory response by increasing the expression of AQP1 and ENaC in lung tissue.
Objective: To investigate the protective effect of intraperitoneal injection of perfluorocarbon(C8 F18)on lung ischemia-reperfusion injury(LIRI) in rats. Methods: A total of 90 healthy and clean Sprague-Dawley rats were randomly divided into blank group, control group, and experimental group. A LIRI model was established in rats. The blank group(NC group) underwent thoracotomy without LIRI treatment. The control group(NS group) received intraperitoneal injection of 0.9% NS at 48 h before thoracotomy and LIRI treatment. The experimental group(PFC group) received intraperitoneal injection of C8 F18 at 48 h before thoracotomy and LIRI treatment. Arterial blood gas parameters and the expression of tumor necrosis factor(TNF-α), lung aquaporin(AQP1), and sodium channel protein(ENaC) were measured in the three groups. Results: There was a significant difference in partial pressure of carbon dioxide(PaCO2) at 2 h between the NS group and the PFC group(P < 0.05), but there were no significant differences in PaCO2 at 4 and 6 h between the two groups(P > 0.05). The PFC group had a significantly lower level of tumor necrosis factor-alpha(TNF-α) in whole blood at each time point compared with the NS group(P <0.05). Among the three groups, the NC group had the highest expression of AQP1. The staining of AQP1 in the NS group was significantly reduced, especially at 2 h, and the NS group had significantly lower expression of AQP1 at each time point compared with the NC group(P < 0.05). The PFC group had significant higher expression of AQP1 compared with the NS group(P < 0.05). Among the three groups, the NC group had the highest expression of ENaC.The staining of ENaC in the NS group was significantly reduced, and the NS group had significantly lower expression of ENaC at each time point compared with the NC group(P < 0.05). There was no significant difference in the expression of ENaC between the PFC group and the NS group(P > 0.05). Conclusion: In the LIRI rat model, intraperitoneal injection of perfluorocarbon has a protective effect on the lung tissue by promoting pulmonary gas exchange — increasing PaO2 in arterial blood and decreasing PaCO2 and by reducing the level of TNF-α in whole blood. At the same time, it can effectively alleviate pulmonary edema and inflammatory response by increasing the expression of AQP1 and ENaC in lung tissue.
引文
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2.Villar J,Blanco J,Manuel Anon J,et al.The alien study:incidence and outcome of acute respiratory distress syndrome in the era of lung protective ventilation[J].Intensive Care Med,2011,37(12):1 932-1 941.
3.Laubach VE,Sharma AK.Mechanisms of lung ischemiareperfusion injury[J].Curr Opin Organ Transplant,2016,21(3):246-252.
4.Weyker PD,Webb C,Kiamanesh D,et al.Lung ischemia reperfusion injury a bench-to-bedide review[J].Semin Cardiothorac Vasc Anesth,2012,17(1):28-43.
5.Sayah DM,Mallavia B,Liu FC,et al.Neutrophil extracellular traps are pathogenic in primary graft dysfunction after lung transplantation[J].Am J Respir Crit Care Med,2015,191(4):455-463.
6.Albertine KH,Soulier MF,Wang Z,et al.Fas and Fas ligand are upregulated in pulmonary oedema fluid and lung tissue of patients with acute lung injury and the acute respiratory distress syndromel[J].Am J Pathol,2002,161(5):1 783-1 796.
7.Guerra Mora JR,Perales Caldera E,Aguilar Leon D,et al.Effects of sildenafil and tadalafil on edema and reactive Oxygen species production in an experimental model of lung Ischemia-Reperfusion injury[J].Transplant Proc,2017,49(6):1 461-1 466.
8.Forgiarini LF,Forgiarini LA,da Rosa DP,et al.N-acetylcysteine administration confers lung protection in different phases of lung ischaemia-reperfusion injury[J].Interact Cardiovasc Thorac Surg,2014,19(6):894-899.
9.Fan YF,Zhang DG,Xiang DK.Delayed protective effect of telmisartan on lung ischemia/reperfusion injury in valve replacement operations[J].Exp Ther Med,2016,12(4):2 577-2 581.
10.Motoyama H,Chen FS,Hijiya K,et al.Plasmin administration during ex vivo lung perfusion ameliorates lung ischemia-reperfusion injury[J].Journal of Heart and Lung Transplantation,2014,33(10):1 093-1 099.
11.Sarkar S,Paswan A,Prakas S.Liquid ventilation[J].Anesth Essays Res,2014,8(3):277-282.
12.Vov der Hardt k,Schoof E,Kandler M,et al.Aerosolized perfluorocarbon sup-presses early pulmonary inflammatory respones in a surfactat-depleted Piglet model[J].Pediatr Res,2002,51(2):177-182.
13.Obraztov VV,Neslund GG,Kornbrust ES,et al.In vitro cellular effects of perfluorochemicals correlate with their lipid solubility[J].Am J Physiol Lung Cell Mol Physiol,2000,278(5):1 018-1 024.
14.Koch T,Ragaller M,Haufe D,et al.Perfluorohexane attenuates proinflamma-tory and procoagulatory response of activated monocytes and alveolar macrop-hages[J].Anesthesiology,2001,94(1):101-109.
15.Rüdiger M,Wissel H,Ochs M,et al.Perfluorocarbons are taken up by isolated type II pneumocytes and influence its lipid synthesis and secretion[J].Crit Care Med,2003,31(4):1 190-1 196.
16.Merry HE,Phelan P,Doaks M,et al.Functional roles of tumor necrosis factor-alpha and interleukin 1-Beta in hypoxia and reoxygenation[J].Ann Thorac Surg,2015,99(4):1 200-1 205.
17.Agre P.Aquaporin water channels:from atomic structure to clinical medicine[J].Acta Pharmacol Sin,2006,27(1):27.
18.樊毫军,刘书盈,张健鹏,等.腹腔注射全氟化碳原液治疗急性肺损伤的可行性探讨[J].中国危重病急救医学,2006,18(8):503-504.
19.Song Y,Yang B,Matthay MA,et al.Role of aquaporin water channels in pleural fluid dynamics[J].Am J Physiol Cell Physiol,2000,279(6):1 744-1 750.
20.Tomassoni D,Bramanti V,Amenta F.Expression of aquaporins 1 and 4 in the brain of spontaneously hypertensive rats[J].Brain Res,2010,1325(14):155-163.
21.Su X,Song Y,Jiang J,et al.The roIe of aquaporin-l(AQPl)pression in amurine model of lipopolysaccharide-induced acute lung injury[J].Respir Physiol Neuobiol,2004,142(1):1-11.
22.Johnson M,Widdicombe J,Allen L,et al.Alveolar epithelial type I cells contain transport proteins and transport Sodium,supporting an active role for type I cells in regulation of lung liquid homeostasis[J].Proc Natl Acad Sci,2002,99(4):1 966-1 971.
23.Canessa CM,Schild L,Buell G.Amiloride-sensitive epithelial Na+Channel is made of three homolous subunits[J].Nature,1994,367(6462):463-466.