西格列汀对2型糖尿病大鼠血清及组织中chemerin水平的影响

发布时间：2018-03-25 04:00

  本文选题：2型糖尿病　切入点：西格列汀　出处：《安徽医科大学》2015年硕士论文

【摘要】：目的研究西格列汀对2型糖尿病(T2DM)大鼠各代谢指标及脂肪因子chemerin水平的影响及其可能的机制,并探讨脂肪因子chemerin与T2DM的关系。方法1.选取年龄4周且体重为(100±20)g的普通级雄性SD大鼠,共35只,在室温为22℃及湿度在55%左右的环境下不限制摄食及饮水喂养。2.造模:将大鼠分为普通饲料喂养组(NC组,n=10)及高脂饲料喂养组(HF组,n=25),12周后一次性腹腔注射链脲佐菌素STZ(用之前溶解在PH=4.5的0.1mmol/L柠檬酸缓冲液中)35 mg/kg于高脂大鼠,7天后测定大鼠空腹血糖,将空腹血糖11.1 mmol/L设定为临界点,超过该临界点的大鼠认为T2DM模型造模成功。3.干预方式:将20只造模成功的大鼠随机分为2组:T2DM对照组(T2DM组,n=10):灌胃2ml生理盐水,西格列汀组(SITA组,n=10):灌胃给予西格列汀10mg/kg/d,溶于2 ml羟甲基淀粉酶中,每日灌胃时间于早9点,持续6周,继续予以普通饲料喂养。4.留取标本:干预前禁食12 h后内眦静脉取血2 ml,干预6周后禁食12 h腹主动脉取血3 ml,血标本静止15 min后,3 200 r/min离心10 min,留取血清置于-80℃冰箱保存。并在冰上操作取出肾周脂肪、肝脏组织及股侧肌肉组织,置于液氮罐中以备提取蛋白质。5.检测指标及方法:干预前后测体重(BW),ELISA法测定血清chemerin、胰岛素(FINS),血糖仪测定空腹血糖(FBG),酶法及选择性沉淀法测定血脂包括总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL),通过公式计算胰岛素抵抗指数(HOMO-IR)与胰岛β细胞功能指数(HOMO-β),Western blot法检测大鼠肾周脂肪、肝脏和肌肉组织chemerin蛋白表达水平。结果1.西格列汀干预前:T2DM大鼠的血清chemerin、BW、FBG、FINS、TC、TG、LDL、HOMO-IR与NC组比较均增高,HOMO-β降低,T2DM组大鼠肾周脂肪、肝脏及肌肉组织的chemerin蛋白表达量增加(P0.05)2.西格列汀干预后,与T2DM组比较,SITA组BW、FBG、FINS、TC、TG、LDL、HOMO-IR及血清chemerin水平降低,HOMO-β升高(P0.05)。SITA组大鼠肾周脂肪、肝脏组织的chemerin蛋白表达量减少(P0.05),肌肉组织中chemerin表达量无变化。3.相关性分析:西格列汀干预前血清chemerin与BW、FBG、FINS、HOMO-IR、TG、TC及LDL呈正相关,干预后血清chemerin与BW、FINS、TG及HOMO-IR呈正相关,血清中chemerin的含量与肾周脂肪中该蛋白的表达量呈正相关(r=0.79,P0.05)。4.多元回归分析:显示西格列汀干预前后TG、FINS及HOMO-IR始终与chemerin独立相关。血清chemerin水平与肾周脂肪chemerin蛋白表达量呈正相关(r=0.79,P0.05)。结论2型糖尿病大鼠血清及脂肪、肝脏、肌肉组织中chemerin水平较非糖尿病大鼠增加,提示在多种代谢性疾病如肥胖、2型糖尿病的发病过程中,脂肪因子chemerin的可能有重要的作用。西格列汀可降低血清、肾周脂肪及肝脏组织中chemerin水平,改善血清学各代谢指标。
[Abstract]:Objective to study the Sig Leo Dean of type 2 diabetes mellitus (T2DM) rats of various metabolic indicators and fat levels of factor Chemerin and its possible mechanism, and to explore the relationship of adipokine Chemerin and T2DM. Methods 1. aged 4 weeks and weighing (100 + 20) ordinary male SD g rats, a total of 35. At room temperature is 22 degrees and the humidity is about 55% under the environment of restricted feeding and drinking feeding.2. rats: the rats were divided into normal diet group (group NC, n=10) and high-fat diet group (group HF, n=25), 12 weeks after intraperitoneal injection of streptozotocin (STZ before the use of 0.1mmol/L dissolved in citrate buffer in PH=4.5 35 mg/kg) in hyperlipidemia rats, 7 days after the determination of fasting blood glucose, fasting blood glucose of 11.1 mmol/L was set as the critical point, more than the critical point that rat T2DM model was successfully built.3. intervention: 20 successful model of large rats were randomly divided into 2 groups: T2DM control group (group T2DM, n=10) 2ml gavage of saline, Sig Leo Dean group (group SITA, n=10): intragastric administration of Sig Leo Dean 10mg/kg/d, dissolved in 2 ml hydroxymethyl amylase, daily gavage time in early 9, for 6 weeks, continue to be fed with normal diet.4. specimens specimen: before intervention after fasting for 12 h angular vein blood 2 ml, 6 weeks after the intervention 12 h fasting blood from the abdominal aorta 3 ml blood samples were still 15 min after 3200 r/min centrifugation for 10 min, the serum -80 in C refrigerator. In the ice and pulled out the perirenal fat and liver tissue and the side muscle tissue in liquid nitrogen tank for extracting detection index and method of protein.5.: measurement of body weight before and after the intervention (BW), serum Chemerin ELISA, insulin (FINS), fasting blood glucose (FBG), serum lipids including total cholesterol enzymatic and selective precipitation method (TC), triglyceride (TG), low density lipoprotein 鐧，

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