蛋白转导Rab GEF1减轻小鼠肝脏缺血再灌注所致的肺损伤  

Intravenous injection of Tat-Rab GEF1 fusion protein attenuates lung injury induced by hepatic ischemia/reperfusion in mice

作　　者：方虹仪 程楠[1] 谢汉镔[1] 罗晨芳[1] FANG Hong-yi;CHENG Nan;XIE Han-bin;LUO Chen-fang(Department of Anesthesiology,Third Affiliated Hospital,Sun Yat-sen University,Guangzhou 510630,China)

机构地区：[1]中山大学附属第三医院麻醉科,广东广州510630

出　　处：《中国病理生理杂志》2019年第2期340-345,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学青年基金资助项目(No.81501938);广东省科技计划项目(No.2014A020212158)

摘　　要：目的:探讨转录反式激活因子-Rab鸟嘌呤核苷酸交换因子1(Tat-RabGEF1)融合蛋白对小鼠肝脏缺血再灌注(IR)所致肺损伤的作用及其可能的机制。方法:选用雄性C57BL/6小鼠24只,按随机数字表法分为假手术(sham)组、IR组和Tat-RabGEF1组,每组8只。采用肝脏缺血90 min再灌注4 h制备肺损伤模型,Tat-Rab-GEF1组于再灌注即刻经尾静脉注射Tat-RabGEF1 (10 mg/kg)。再灌注4 h后处死小鼠,收集支气管肺泡灌洗液(BALF),采用ELISA法检测BALF中肿瘤坏死因子α(TNF-α)、白细胞介素6 (IL-6)和白细胞介素1β(IL-1β)的含量;取肺组织进行病理学检测,测定湿/干重比、β-氨基己糖苷酶(β-Hex A)和髓过氧化物酶(MPO)水平,并采用Western blot法测定肺组织类胰蛋白酶的表达水平。结果:(1)肝缺血90 min再灌注4 h肺组织病理损伤明显,湿/干重比增高,Tat-RabGEF1尾静脉注射明显减轻肺脏病理损伤。(2)与IR组比较,Tat-RabGEF1组BALF中TNF-α、IL-6和IL-1β浓度,以及肺组织β-Hex A、MPO水平和类胰蛋白酶表达显著降低(P <0.05)。结论:体内蛋白转导Rab GEF1减轻肝IR所致的肺损伤,减少肺组织炎症反应和细胞因子水平,其机制可能与抑制肥大细胞激活有关。AIM:To investigate the effect of trans-activator of transcription-Rab guanine nucleotide exchange factor 1(Tat-RabGEF1)fusion protein by intravenous injection on hepatic ischemia/reperfusion(IR)-induced lung injury,and its underlying mechanism.METHODS:Adult male C57BL/6 rats(n=24)were randomized into 3 groups with 8 rats each:sham group,IR group,and Tat-RabGEF1 group.IR was induced by occlusion of the vessels in the hepatic pedicle for 90 min followed by 4 h of reperfusion.The Tat-RabGEF1 protein was intravenously administered right after reperfusion through the caudal vein.The levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and interleukin-1β(IL-1β)in bronchoalveolar lavage fluid(BALF)were assessed by ELISA.The lung histopathological injury score and lung wet/dry weight ratio were evaluated,and the levels of β-hexosaminidase A(β-Hex A)and myeloperoxidase(MPO)in the lung tissues were also measured.The expression of tryptase in the lung was determined by Western blot.RESULTS:The pathological changes after ischemia for 90 min followed by reperfusion for 4 h were significant,along with increased lung wet/dry weight ratio and lung injury score.Intravenous injection of Tat-RabGEF1 protein effectively alleviated the lung pathological injury(P<0.05).Compared with IR group,lower levels of TNF-α,IL-6 and IL-1β in BALF,and lower expression of β-Hex A,MPO and tryptase in the lung tissues in Tat-RabGEF1 group were observed(P<0.05).CONCLUSION:Intravenous injection of Tat-RabGEF1 protein attenuates the lung injury after IR,and its mechanism may be related with inhibition of mast cell activation,reduced releases of β-Hex A and tryptase,and decreased levels of TNF-α,IL-6,IL-1β and MPO.

关 键 词：肝缺血再灌注 肺损伤 肥大细胞 Tat-RabGEF1融合蛋白 炎症 

分 类 号：R563[医药卫生—呼吸系统] R363[医药卫生—内科学]