自噬在糖尿病大鼠心肌缺血再灌注所致急性肺损伤中的作用机制研究  

Mechanism of autophagy in acute lung injury caused by myocardial ischemia-reperfusion in diabetic rats

作　　者：陈婷婷[1] 戴晓雯[1] CHEN Ting⁃ting;DAI Xiao⁃wen(Department of Anesthesiology,the First Affiliated Hospital of Xinjiang Medical University,Urumqi,Xinjiang 830054,China)

机构地区：[1]新疆医科大学第一附属医院麻醉科,新疆乌鲁木齐830054

出　　处：《热带医学杂志》2022年第6期798-804,895,共8页Journal of Tropical Medicine

基　　金：新疆维吾尔自治区自然科学基金(2019D01C324)

摘　　要：目的探讨肺自噬状态及其在心肌缺血再灌注糖尿病大鼠肺组织氧化应激和炎症反应中的作用,为疾病预防及靶向治疗提供依据。方法将雄性SD大鼠分为假手术组:对照组、糖尿病组;以及心肌缺血再灌注组:对照组、糖尿病组,每组10只。通过单次腹腔注射链脲佐菌素(STZ)(60 mg/kg)诱导糖尿病模型,假手术组中的对照组大鼠注射相同体积的溶剂对照(p H 4.5,柠檬酸盐缓冲液)。糖尿病诱导后8周,将心肌缺血再灌注组中的糖尿病组大鼠麻醉并通过闭塞左冠状动脉前降支(LAD)30 min,然后再灌注120 min来实现心肌缺血再灌注;心肌缺血再灌注组中的对照组大鼠在没有结扎的情况下接受了相同的手术处理;糖尿病组大鼠在缺血前用3-甲基腺嘌呤(3-MA)预处理30 min或静脉注射雷帕霉素(Rap)15 min。再灌注结束时,采用2,3,5-三苯基氯化四唑(TTC)染色测量心肌梗塞大小;采用苏木素伊红染色检测肺组织损伤情况;采用酶联免疫吸附法检测肺泡灌洗液中炎症因子[肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、IL-8]表达情况;进一步采用免疫印迹法检测自噬相关蛋白(LC3-II、LC3-I、Beclin-1)表达情况。结果假手术组中,与对照组大鼠比较,STZ诱导的糖尿病组大鼠进水量、摄食量显著增加,血糖含量显著升高,体重显著减轻,差异均有统计学意义(P均<0.05)。假手术组中对照组和糖尿病组大鼠肺切片未见明显病理改变。而缺血再灌注组大鼠心肌损伤后,肺组织染色中出现明显水肿、血液渗出及相关炎症细胞浸润的现象,呈现急性肺损伤(ALI)的表现,这些改变在糖尿病组中更为严重;糖尿病组大鼠肺损伤评分显著高于对照组,差异有统计学意义(P<0.05)。糖尿病组大鼠中LC3-II/LC3-I比值及Beclin-1表达显著低于对照组,差异均有统计学意义(P均<0.05),心肌缺血再灌注损伤进一步降低,LC3的情况同样。与对照组大鼠比较,接受�Objective To investigate the state of pulmonary autophagy and its role in oxidative stress and inflammatory response in lung tissue of diabetic rats with myocardial ischemia-reperfusion,and to provide evidence for disease prevention and targeted therapy.Methods Male SD rats were divided into sham-operation group:control group,diabetic group;and myocardial ischemia-reperfusion group:control group,diabetic group;10 rats in each group.The diabetes model was induced by a single intraperitoneal injection of streptozotocin(STZ)(60 mg/kg),and control rats in the sham-operation group were injected with the same volume of solvent control(pH 4.5,citrate buffer).Eight weeks after the induction of diabetes,the diabetic rats in the myocardial ischemia-reperfusion group were anesthetized and the left anterior descending coronary artery(LAD)was occluded for 30 min,followed by reperfusion for 120 min to achieve myocardial ischemia reperfusion;The control rats in the myocardial ischemia-reperfusion group received the same surgical treatment without ligation;the diabetic rats were treated with 3-methyladenine(3-MA)for 30 min before ischemia or intravenous injection of Rapa Mycin(Rap)for 15 min.At the end of reperfusion,2,3,5-triphenyl-2H-tetrazolium chloride(TTC)staining was used to measure the size of myocardial infarction;hematoxylin eosin staining was used to detect lung tissue damage;enzyme-linked immunosorbent assay was used to detect the expression of inflammatory factors[tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),IL-8]in bronchoalveolar lavage fluid(BALF);further,Western blotting was used to detect autophagy-related protein(LC3-II,LC3-I,Beclin-1)expression.Results In the sham-operation group,compared with the control group,the STZ-induced diabetic group showed a significant increase in water intake and food consumption,a significant increase in blood sugar content,and a significant reduction in body weight,with statistically significant differences(P<0.05).In the sham-operation group,no obvious pathological change

关 键 词：糖尿病 自噬 肺损伤 缺血再灌注 

分 类 号：R587.1[医药卫生—内分泌]