2型糖尿病Angptl4及炎症因子水平与动脉粥样硬化的关系研究

发布时间：2018-05-31 10:35

本文选题：2型糖尿病 + 动脉粥样硬化　；参考：《苏州大学》2016年硕士论文

【摘要】：【目的】建立小鼠2型糖尿病(type 2 diabetes mellitus,T2DM)模型、动脉粥样硬化(atherosclerosis,AS)模型以及2型糖尿病合并动脉粥样硬化(T2DM+AS)模型,探讨2型糖尿病血管生成素样蛋白4(Angiogenin-like protein 4,Angptl4)、炎症因子与动脉粥样硬化的关系。【方法】将20只ApoE-/-小鼠随机分为对照组、2型糖尿病组(T2DM组)、动脉粥样硬化组(AS组)、2型糖尿病合并动脉粥样硬化组(T2DM+AS组)。普通饲料喂养对照组,高脂饲料喂养建立AS组,烟酰胺+链脲佐菌素+普通饲料建立T2DM组,烟酰胺+链脲菌素+高脂饮食建立T2DM+AS组。对各组ApoE-/-小鼠动脉以及胰腺进行常规及病理学检测;酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)、化学发光免疫分析法(chemiluminescent immunoassay,CLIA)等方法检测血清中Angptl4、血糖、胰岛素、胰岛素抵抗指数(Homeostasis model assessment-IR,HOMA-IR)、甘油三酯(Triacylglycerol,TG)、血浆纤溶酶原激活物抑制物1(plasma plasminogen activator inhibitor-1,PAI-1)、低密度脂蛋白(low density lipoprotein cholesterin,LDL-c)和炎性因子肿瘤坏死因子α(Tumor Necrosis Factorα,TNF-α),转化生长因子β1(Transforming growth factorβ1,TGF-β1)和C反应蛋白(C Reactive Protein,CRP)的含量或浓度;免疫荧光方法分析平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)在各组小鼠的动脉中表达的变化;逆转录-聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)和蛋白质印迹(Western Blot)方法检测Angptl4、α-SMA、血管内皮生长因子、TNF-α、TGF-β1和CRP在各个模型小鼠中的表达变化。【结果】1)血清生化检测显示:模型组(T2DM组、AS组、T2DM+AS组)Angptl4的含量均低于对照组,T2DM+AS组中含量最低;与对照组相比,TNF-α和CRP在AS组和T2DM+AS组中有显著升高,TGF-β1则显著降低,且T2DM+AS组中变化较AS组更加明显;各模型组中胰岛素水平均显著降低,T2DM组中HOMA-IR升高,T2DM+AS组中HOMA-IR升高更加明显;TG//PAI-1/LDL-c在各模型组中均有明显升高,且T2DM+AS组中比T2DM组和AS组变化更加明显。2)与对照组相比,T2DM组的动脉内膜略有狭窄,有斑点出现,有堆叠现象;AS组的动脉内膜同样也有斑点、内膜狭窄和堆叠现象,且中膜有增厚,α-SMA表达量升高;T2DM+AS组的动脉内膜出现斑块,中膜厚度增加较明显,α-SMA表达显著升高。3)胰腺的HE染色表明,与对照组相比,AS组的胰腺相对正常,但有炎性细胞浸润;T2DM组的胰腺小结间隔不明显,腺泡萎缩形成空泡;T2DM+AS组的胰腺小叶间隔不明显,腺泡萎缩形成空泡并有炎性细胞浸润。4)RT-PCR和Western blot结果均表明,与对照组相比,Angptl4在AS组和T2DM组中表达明显降低,且AS组要低于T2DM组,T2DM+AS组中表达最低。与对照组相比,T2DM组中TNF-α,TGF-β1和CRP的表达没有明显变化;AS组中TNF-α和CRP的表达升高,TGF-β1的表达则明显下调,T2DM+AS组中TNF-α和CRP的表达显著高于AS组,TGF-β1的表达则显著低于AS组。【结论】1)Angptl4在2型糖尿病和动脉粥样硬化中的表达降低,并且与糖代谢和脂代谢异常有关;2)Angptl4可能通过对炎症因子的调节延缓2型糖尿病动脉粥样硬化的发展,从而在2型糖尿病动脉粥样硬化的发生和发展中起保护作用。
[Abstract]:[Objective] to establish the model of type 2 diabetes (type 2 diabetes mellitus, T2DM), the model of atherosclerosis (atherosclerosis, AS) and the model of type 2 diabetes with atherosclerotic (T2DM+AS), and explore the angiopoietin like protein 4 (Angiogenin-like protein 4, Angptl4) of type 2 diabetes, and the relationship between the inflammatory factors and atherosclerosis. [method] 20 ApoE-/- mice were randomly divided into control group, type 2 diabetes group (group T2DM), atherosclerosis group (group AS), type 2 diabetes with atherosclerosis group (group T2DM+AS). Normal diet feeding control group, high fat diet set up AS group, nicotinamide + streptozotocin + ordinary feed set up T2DM group, nicotinamide + streptozotocin T2DM+AS group was established with high fat diet. Routine and pathological examination of the arteries and pancreas of ApoE-/- mice in each group; enzyme linked immunosorbent assay (enzyme linked immunosorbent assay, ELISA), chemiluminescent immunoassay (chemiluminescent immunoassay, CLIA) and other methods for detecting Angptl4, blood sugar, insulin, and insulin resistance index in serum (Homeostasis model assessment-IR, HOMA-IR), triglyceride (Triacylglycerol, TG), plasma plasminogen activator inhibitor 1 (plasma plasminogen activator inhibitor-1, PAI-1), low density lipoprotein (low) and inflammatory factor alpha (alpha), transforming growth factor alpha. The content or concentration of long factor beta 1 (Transforming growth factor beta 1, TGF- beta 1) and C reactive protein (C Reactive Protein, CRP), and the changes in the expression of smooth muscle actin (alpha -smooth muscle actin, alpha) in the arteries of each group of mice by immunofluorescence; Action, RT-PCR) and Western blot (Western Blot) method were used to detect the expression of Angptl4, alpha -SMA, VEGF, TNF- a, TGF- beta 1 and CRP in each model mice. [results] 1) serum biochemical tests showed that the content of the model group (T2DM group, AS group, T2DM+AS group) was lower than that in the control group. Compared with the group, TNF- alpha and CRP were significantly increased in group AS and T2DM+AS, and TGF- beta 1 decreased significantly, and the changes in T2DM+AS group were more obvious than those in the AS group. The levels of insulin in each model group were significantly decreased, HOMA-IR in T2DM group increased and HOMA-IR increased in T2DM+AS group, and TG//PAI-1/LDL-c was significantly increased in each model group. The changes in the +AS group were more obvious than that of the T2DM group and the AS group. Compared with the control group, the intima of the arterial intima in the T2DM group was slightly narrow, there were spots and stacked. The intima of the arterial intima in the AS group also had spots, intima stenosis and stacking phenomenon, and the middle membrane was thickened and the expression of alpha -SMA increased; the intima of the arterial intima appeared in the T2DM+AS group and the thickness of the middle membrane was increased. The HE staining of the pancreas was significantly higher in.3) HE staining of the pancreas showed that the pancreas of the AS group was relatively normal, but there was inflammatory cell infiltration, the pancreatic nodule interval was not obvious in the T2DM group, the alveolus atrophied to form vacuoles, the pancreatic interlobular septum in the T2DM+AS group was not obvious, the vesicles formed vacuoles and the inflammatory infiltration of.4) in the T2DM group, and Wes RT-PCR and Wes. Tern blot results showed that, compared with the control group, the expression of Angptl4 in the AS group and the T2DM group was significantly lower, and the AS group was lower than the T2DM group, and the expression in the T2DM+AS group was the lowest. Compared with the control group, the TNF- a, TGF- beta 1 and the expression of CRP, the expression of TGF- beta and CRP in the T2DM group was not significantly changed. The expression of TNF- alpha and CRP was significantly higher than that in group AS, and the expression of TGF- beta 1 was significantly lower than that in group AS. [Conclusion] 1) the expression of Angptl4 in type 2 diabetes and atherosclerosis is reduced, and it is associated with abnormal glucose metabolism and lipid metabolism; 2) Angptl4 may delay the development of atherosclerosis in type 2 diabetes by regulating the inflammatory factors. It plays a protective role in the occurrence and development of atherosclerosis in type 2 diabetes.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R587.2

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