大鼠肢体缺血再灌注急性肺损伤细胞自噬及其相关蛋白的表达  

Expression of autophagy associated proteins in acute lung injury induced by remote limb ischemia reperfusion in rats

作　　者：李跃兵[1,2] 康于庆[1] 叶靖[1] 赵振龙[1] 林春水[1] 秦再生[1] 古妙宁[1] 

机构地区：[1]南方医科大学附属南方医院麻醉科,广州510515 [2]浙江中医药大学附属第二医院麻醉科,杭州310005

出　　处：《重庆医学》2014年第32期4280-4282,4287,共4页Chongqing medicine

摘　　要：目的研究大鼠双下肢缺血再灌注所致急性肺损伤(LIR-ALI)肺组织细胞早期自噬及其相关蛋白的表达。方法采用LIR-ALI模型,12只雄性SD大鼠,以随机数字表法分为假手术(Sham)组,肢体缺血再灌注(I/R)组,每组6只。采用ELISA检测血清白乳酸脱氢酶(LDH)和免疫荧光法检测肺组织病理变化,利用RT-PCR检测各组肺组织Beclin1和Atg5mRNA表达及Western blot检测LC3蛋白的表达。结果与Sham组比较,I/R组LDH值升高,显示大鼠LIR-ALI模型造模成功且肺组织免疫荧光特异性标记抗体高表达(P<0.01)。采用2-△△CT法分析肺组织Beclin1和Atg5mRNA相对表达量,在I/R组较Sham组升高(P<0.01);Western blot法检测显示,I/R组较Sham组LC3-Ⅰ、LC3-Ⅱ高表达、LC3-Ⅱ/GAPDH亦明显升高(P<0.01)。结论大鼠LIR-ALI后激活肺组织细胞自噬及其相关蛋白高表达。Objective To explore the early expression of autophagy associated proteins in lung tissues in acute lung injury induced by remote limb ischemia reperfusion（LIR-ALI）in Rats.Methods Twelve adult male SD rats weighting 220-250 g were made LIR-ALI models and divided randomly into two groups（6in each group）：Sham operation group and ischemia-reperfusion（I/R）group.The rats were anesthetized and the tissues of lung were removed at the end of 4hours of reperfusion after 3hours of ischemia.The serum lactate dehydrogenase（LDH）were detected with ELISA and the pathological changes of lung tissues were observed by immunofluorescence techniques;the expression of Beclin1 protein and Atg5 mRNA in the lung tissues were detected by reverse transcription PCR（RT-PCR）.Microtubules associated protein light chain 3（LC3）in the lung tissues were detected by Western blot test.Results Compared with Sham group,the level of serum LDH in I/R group were very higher than Sham group（P〈0.01）,which showed that the rats models of LIR-ALI were established.The expression of specific antibody,Beclin1 protein,Atg5mRNA and LC3-Ⅰ,LC3-Ⅱ and the ratio of LC3-Ⅱ/GAPDH in lung tissues were all very higher in I/R group than in Sham group（P〈0.01）.Conclusion ALI can be induced by LIR and the expression of autophagy associated proteins in lung tissues in LIR-ALI model is very higher in rats.

关 键 词：呼吸窘迫综合征 成人 自噬 肢体缺血再灌注 BECLIN1 LC3 

分 类 号：R563.8[医药卫生—呼吸系统]