STZ诱导联合高脂喂养LDLR^(-/-)小鼠糖尿病动脉粥样硬化模型的构建及评价  

Construction and evaluation of diabetic atherosclerosis model in LDLR^(-/-)mice induced by STZ and fed with high-fat diet

作　　者：王丽 高金宁[2] 陈湘云 姜沛青 李梅 步芳芳 张梅[3] 姚桂华 孙慧 WANG Li;GAO Jinning;CHEN Xiangyun;JIANG Peiqing;LI Mei;BU Fangfang;ZHANG Mei;YAO Guihua;SUN Hui(Department of Cardiology,Qilu Hospital(Qingdao),Shandong University,Qingdao,Shandong 266053,China;Academic Affairs Office of Qingdao University,Qingdao,Shandong 266000,China;Department of Cardiology,Qilu Hospital,Shandong University,Jinan,Shandong 250012,China)

机构地区：[1]山东大学齐鲁医院(青岛)心内科,山东省青岛市266053 [2]青岛大学教务处,山东省青岛市266000 [3]山东大学齐鲁医院心内科,山东省济南市250012

出　　处：《中国动脉硬化杂志》2024年第11期985-993,共9页Chinese Journal of Arteriosclerosis

基　　金：国家自然科学基金面上项目(81970282);青岛市临床重点专科建设项目(QDZDZK-2022008)。

摘　　要：[目的]构建糖尿病动脉粥样硬化小鼠模型,并研究该小鼠模型中糖尿病促动脉粥样硬化的病理特点。[方法]8周龄雄性LDLR^(-/-)小鼠50只,普脂饮食适应性喂养2周后改为高脂饮食,并随机分为两组,糖尿病动脉粥样硬化组采用小剂量链脲佐菌素(STZ)腹腔注射5天造模,动脉粥样硬化组同时给予柠檬酸盐缓冲液注射。多次检测两组小鼠体质量、血糖、血脂,于23周龄糖耐量检测后安乐处死,采用HE染色和油红O染色检测小鼠全长主动脉和主动脉根部动脉粥样硬化,免疫组织化学染色检测斑块内CD4、α-平滑肌肌动蛋白(α-SMA)、巨噬细胞含EGF样模块黏蛋白样激素受体1(EMR1)、单核细胞趋化蛋白1(MCP-1)、NOD样受体蛋白3(NLRP3)、血管细胞黏附分子1(VCAM-1)、基质金属蛋白酶2(MMP-2)、组织金属蛋白酶抑制物1(TIMP-1),Western blot检测α-SMA、CD4、肿瘤坏死因子α(TNF-α)、NLPR3、细胞间黏附分子1(ICAM-1)、Ⅰ型和Ⅲ型胶原。[结果]与动脉粥样硬化组相比,糖尿病动脉粥样硬化组小鼠体质量降低,总胆固醇(TC)、甘油三酯(TG)和低密度脂蛋白胆固醇(LDLC)水平升高,高密度脂蛋白胆固醇(HDLC)水平降低,糖耐量降低(P<0.05)。与动脉粥样硬化组相比,糖尿病动脉粥样硬化组动脉粥样硬化斑块分布弥漫且面积增加,其中脂质、T细胞、巨噬细胞、平滑肌细胞、Ⅰ型和Ⅲ型胶原含量增加(P<0.05);血管组织中TNF-α、MCP-1、MMP-2、NLRP3、ICAM-1、VCAM-1蛋白含量增加,而TIMP-1蛋白含量降低,MMP2/TIMP-1比值升高(P<0.05)。[结论]通过STZ诱导联合高脂饮食,可将LDLR^(-/-)小鼠成功构建糖尿病动脉粥样硬化模型,并能体现糖尿病促动脉粥样硬化的斑块组成及炎症特点,可作为一种相对理想的研究糖尿病大血管病变的病理模型。Aim To construct a diabetic atherosclerosis mouse model and study the pathological characteristics of diabetic atherosclerosis.Methods Fifty 8-week-old male LDLR^(-/-)mice were fed with standard diet for 2 weeks and then changed to high-fat diet,they were randomly divided into two groups.The diabetic atherosclerosis group was given intraperitoneal injection of low dose streptozotocin(STZ)for 5 days continuouly to establish the model,and the atheroscle-rosis group was given citrate buffer injection at the same time.The body mass,blood glucose and blood lipids of the mice in the two groups were detected for many times.At the age of 23 weeks,the mice were euthanized after glucose tolerance test.HE staining and oil red O staining were used to detect the gross and aortic root atherosclerosis,immunohistochemical staining was used to detect CD4,α-smooth muscle actin(α-SMA),EGF-like module-containing mucin-like hormone re-ceptor-like 1(EMR1),monocyte chemotactic protein-1(MCP-1),NOD-like receptor protein 3(NLRP3),vascular cell adhesion molecule-1(VCAM-1),matrix metalloproteinase-2(MMP-2)and tissue inhibitor of metalloproteinase-1(TIMP-1),Western blot was used to detectα-SMA,CD4,tumor necrosis factor-α(TNF-α),NLPR3,intercellular adhesion molecule-1(ICAM-1),and typeⅠandⅢcollagen.Results Compared with the atherosclerosis group,the body mass decreased,the levels of total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDLC)increased,and the levels of high density lipoprotein cholesterol(HDLC)decreased(P<0.05)in the diabetic atherosclerosis group.Compared with the atherosclerosis group,the distribution of atherosclerotic plaques was diffuse and the area was increased in the diabetic atherosclerosis group,and the contents of lipids,T cells,macrophages,smooth muscle cells,typeⅠandⅢcolla-gen were increased(P<0.05);the protein levels of TNF-α,MCP-1,MMP-2,NLRP3,ICAM-1 and VCAM-1 in vascular tissues were increased,while the content of TIMP-1 were decreased and MMP2/TIMP-1 were increased(P<0.05).Co

关 键 词：链脲佐菌素 高脂饮食 糖尿病 动脉粥样硬化 LDLR^(-/-)小鼠 

分 类 号：R363[医药卫生—病理学] R5[医药卫生—基础医学]