【作者】 任冰稳；

【导师】 叶平；

【作者基本信息】 中国人民解放军军医进修学院 ， 老年医学心血管内科， 2010， 博士

【摘要】 目的：代谢紊乱及相关疾病已成为严重危害人类健康的主要疾病,其发病机制目前尚不清楚。流行病学提示代谢紊乱的发病率增加与人们精神应激增加和活动减少关系密切。本试验以束缚应激大鼠为模型,1、观察束缚应激对大鼠血脂、血糖的影响及肝脏、脂肪、肌肉和心肌等重要代谢和应激器官PPARs的变化,初步探讨PPARs在应激代谢中的作用；2、观察TZDs类降糖药吡格列酮对束缚应激大鼠代谢的干预作用及作用机制,为代谢紊乱的治疗提供试验依据。方法：1、雄性Wister大鼠,分为正常对照组(C)、束缚1周组(R1)、束缚2周(R2)和束缚4周组(R4),测定各组大鼠血糖、TG、TC、HDL-C、LDL-C、FFA、血清皮质酮、胰岛素和去甲肾上腺素。以RT-PCR和western blot技术测定各组大鼠肝脏、肌肉、心肌和脂肪中PPARα、PPARβ／δ、PPARγ、GLUT-4、LPL和CPT-I(肝脏为L-CPT,肌肉、心肌和脂肪为M-CPT)mRNA和蛋白的表达。2、雄性Wister大鼠,分为正常对照组(C2)、束缚应激组(R)、束缚应激加小剂量吡格列酮治疗组(P10,10mg／kg·d)和束缚应激加大剂量吡格列酮治疗组(P20,20mg／kg·d)。应激大鼠给予4周束缚应激后测定各组大鼠血糖、TG、TC、HDL-C、LDL-C、FFA、血清皮质酮、胰岛素和去甲肾上腺素。以RT-PCR和western blot技术测定各组大鼠脂肪中PPARγ、GLUT-4和LPL mRNA和蛋白的表达。结果：1、与C组大鼠相比, R1、R2、R4组大鼠血糖、TG、HDL-C、FFA、皮质酮、去甲肾上腺素和胰岛素明显升高(血糖R1、R2、R4组,HDL-C R2、R4组去甲肾上腺素R4组,胰岛素R1组P<0.05；其余P<0.01),胰岛素敏感指数降低(P<0.01),TC改变不明显(P>0.05),只有R1组LDL-C降低(P<0.05)。2、与R组大鼠相比,P20组大鼠血糖明显降低(P<0.05),两吡格列酮治疗组TG、FFA、胰岛素水平明显降低(P<0.01),胰岛素敏感性增加(P<0.01),TC、LDL-C、HDL-C、皮质酮、去甲肾上腺素变化不明显(P>0.05)。与C2组大鼠相比,P20组大鼠HDL-C明显升高(P<0.05),两吡格列酮治疗组HDL-C、皮质酮、去甲肾上腺素皮质酮、去甲肾上腺素升高(P<0.01),LDL-C降低(P10组P<0.05,P20组P<0.01),血糖、TG、TC、FFA、胰岛素、胰岛素敏感指数无明显差别(P>0.05)。3、与C1组大鼠相比,R1、R2、R4组大鼠肝脏中PPARα、L-CPTmRNA和蛋白表达明显升高(P<0.01),PPARβ／δ、PPARγ、GLUT-4 mRNA和蛋白表达明显降低(PPARβ／δ蛋白R1组P<0.05,其余P<0.01),LPL mRNA和蛋白表达变化不明显(P>0.05)；4、与C1组大鼠相比,R1、R2、R4组大鼠肌肉中PPARα、PPARβ/δ、PPARγ、GLUT-4、LPL和M-CPTmRNA和蛋白表达明显降低(M-CPTmRNA R1组、PPARβ／δ蛋白R1组P<0.05,其余P<0.01)；5、与C1组大鼠相比,R1、R2、R4组大鼠心肌中PPARα、LPL和M-CPTmRNA和蛋白表达明显升高(PPARamRNA R2组、M-CPT mRNA R2组、M-CPT蛋白R4组P<0.05,其余P<0.01)；PPARγ、GLUT-4mRNA和蛋白表达明显降低(P<0.01),PPARβ/δmRNA和蛋白表达变化不明显(P>0.05)；6、与C1组大鼠相比,R1、R2、R4组大鼠脂肪中PPARα、PPARβ/δ、PPARγ2、GLUT-4、LPL、M-CPTmRNA和蛋白表达明显降低(GLUT-4mRNA R1组、LPLmRNA R2和R4组、LPL蛋白R1和R4组P<0.05,其余P<0.01)；7、与R组大鼠相比,两吡格列酮治疗组大鼠脂肪中PPARγ、GLUT-4、LPLmRNA和蛋白表达明显升高(PPARγmRNA、GLUT-4mRNA均Plo组P<0.05,P20组P<0.01；其余P<0.01)。与C2大鼠相比,P10组大鼠脂肪中LPLmRNA、PPARγ、GLUT-4和LPL蛋白表达仍明显降低(P<0.05),Plo组大鼠脂肪中PPARγ和GLUT-4mRNA、P20组大鼠脂肪中PPARγy、GLUT-4、LPLmRNA和蛋白表达无明显差别(P>0.05)结论：1、束缚应激大鼠存在代谢紊乱,以血糖和TG升高为主；2、束缚应激可影响大鼠肝脏、肌肉、心肌和脂肪PPARs及其调控基因mRNA和蛋白表达发生变化,这些变化可能与束缚应激导致的代谢紊乱有关；3、吡格列酮可通过增加脂肪中PPARγ及其调控基因表达和活性,改善束缚应激大鼠的代谢紊乱。更多还原

【Abstract】 Objective:1.To investigate the changes of blood glucose and lipid of rats under restraint stress.2.To detct the expressions of PPARs in the liver、muscle、myocardium and fat in the rats under restraint stress.3、To explore the effects of pioglitazone on the metabolism of rats under restraint stress and its mechanism。Methods:1.Male Wistar rats were divided into four groups:control and rats under 1 week (R1)、2 weeks (R2) or 4 weeks (R4) restraint stress. Blood glucose、TG、TC、HDL-C、LDL-C、FFA、corticosterone、noradrenaline and insulin were detected。The gene and protein expressions of PPARs、GLUT-4、LPL and CPT in the liver、muscle、myocardium and fat were evaluated by RT-PCR and western blot.2.Male Wistar rats were divided into four groups:control、rats under restraint stress、rats given both restraint stress and 10mg/kg-d pioglitazone (P10) and rats given both restraint stress and 20mg/kg-d pioglitazone (P20). Blood glucose、TG、TC、HDL-C、LDL-C、FFA、corticosterone、noradrenaline and insulin were detected. The gene and protein expressions of PPARγ、GLUT-4 and LPL in the fat were evaluated by RT-PCR and western blot. Probability values of 0.05 was considered as significant.Results:1.Compared with C1 group rats, blood glucose、TG、HDL-C、FFA、corticosterone、noradrenaline and insulin in the rats of R1、R2 and R4 group increased significantly, IAI decreased, LDL-C in R1 group also decreased, TC has no significant change.2.Compared with R group rats, blood glucose in the rats of P20 group decreased significantly, blood TG、FFA and insulin in the rats of P10 and P20 group decreased, IAI increased, blood TC、LDL-C、HDL-C、corticosterone and noradrenaline have no significant change.Compared with C2 group rats, blood HDL-C in the rats of P20 group increased significantly, blood HDL-C、corticosterone and noradrenaline in the rats of P10 and P20 group all increased, LDL-C decreased, blood glucose、TG、TC、FFA、 insulin and IAI have no significant change.3.The expressions of PPARα、L-CPT mRNA and protein in the liver of rats of R1、R2 and R4 group were all significantly higher than that of control group, PPARβ/δ、PPARγ、GLUT-4 mRNA and protein were all significantly lower, the expressions of LPL mRNA and protein have no signigfican change.4.The expressions of PPARα、PPARβ/δ、PPARγ、GLUT-4、LPL、M-CPT mRNA and protein in the muscle of rats of R1、R2 and R4 group were all significantly lowered than that of control group.5.The expressions of PPARα、LPL、M-CPT mRNA and protein in the myocardium of rats of R1、R2 and R4 group were significantly higher than that of control group, PPARγ、GLUT-4 mRNA and protein were significantly lower, the expressions of PPARβ/δmRNA and protein have no signigfican change.6.The expressions of PPARα、PPARβ/δ、PPARγ、GLUT-4、LPL、M-CPT mRNA and protein in the fat of rats of R1、R2 and R4 group were all significantly lower than that of control group.7.Compared with R group rats, the expressions of PPARγ、GLUT-4、LPL mRNA and protein in the fat of rats of P10 and P20 group decreased.Compared with C2 group rats, the expressions of LPL mRNA and PPARγ、GLUT-4、LPL protein in the fat of rats of P10 group still decreased, the expression of LPLmRNA、PPARγ、GLUT-4、LPL protein in P10 group and LPL mRNA、PPARγ、GLUT-4、LPL protein in P20 group have no signigfican change。Conclusion:1.Rats under restraint stress have metabolic abnormality, with higer blood glucose and TG.2.The gene and protein expressions of PPARs in the liver、muscle、myocardium and fat were changed in the rats under restraint stress, which may have relationship with metabolic abnormality.3.Rats under restraint stress by pioglitazone treatment can inhibit metabolic abnormality by increasing the the expressions of PPARγ、GLUT-4、LPLmRNA and protein in the fat of rats.更多还原