不同运动时间对胰岛素抵抗大鼠尾加压素Ⅱ及其受体影响的研究
摘要
目的:尾加压素Ⅱ(UrotensinⅡ,UⅡ)是迄今所知作用最强的缩血管活性物质,与胰岛素抵抗密切相关。代谢综合征是目前影响人类健康及生活质量的主要慢性疾病之一。运动作为能够有效改善代谢综合征的重要手段,其作用机制尚未完全阐明。本研究通过检测不同运动时间对胰岛素抵抗大鼠心肌、主动脉、骨骼肌组织中尾加压素Ⅱ及其受体的影响,为代谢综合征正科学运动处方的制定提供一定理论基础。
方法:采用高脂饲料喂养方法建立胰岛素抵抗大鼠模型。随机将正常大鼠分为安静正常对照组(N0),正常+游泳1小时组(N1);正常+游泳2小时组(N2);正常+游泳力竭组(N3);每组10只。高脂膳食致胰岛素抵抗组大鼠随机分为四组:高脂膳食安静组(G0);高脂膳食+游泳1小时组(G1);高脂膳食+游泳2小时组(G2);高脂膳食+游泳力竭组(G3)每组15只。采用全自动生化分析仪测试血脂、血糖;放射免疫法测试胰岛素、血浆尾加压素Ⅱ;免疫组织化学半定量法测试心肌、主动脉、骨骼肌尾加压素Ⅱ及其受体含量;实时荧光定量PCR法测定各种组织中尾加压素Ⅱ及其受体mRNA表达。
结果:高脂膳食组大鼠血脂、空腹血糖、胰岛素均明显升高而胰岛素敏感指数下降。长时间游泳运动能够降低正常大鼠和胰岛素抵抗大鼠血浆UⅡ水平,一次性力竭游泳运动能够提高血浆UⅡ水平。长时间游泳运动能够降低正常大鼠心肌、主动脉、骨骼肌UⅡ及GPR14含量,但是它对UⅡmRNA及GPR14mRNA无显著性影响。长时间游泳运动能够降低胰岛素抵抗大鼠心肌、主动脉、骨骼肌UⅡ及GPR14含量,而且对各个组织局部UⅡmRNA及GPR14mRNA产生显著影响。在正常大鼠和胰岛素抵抗大鼠,一次性力竭运动都能够升高血浆中UⅡ水平,而且各组织UⅡ和其受体的含量均有升高趋势。
结论:本高脂饲料配方建立胰岛素抵抗大鼠模型是可行的。运动可以通过影响血浆中UⅡ来参与机体调节。有氧运动通过UⅡ及GPR14参与对正常及胰岛素抵抗大鼠局部组织的调节可能主要发生在翻译水平或者蛋白质的结构改变环节而不是在转录水平。UⅡ和GPR14在心肌、主动脉、骨骼肌分布变化的同步性提示UⅡ主要通过与其受体GPR14结合,在局部通过自分泌或者旁分泌途径发挥作用。UⅡ也参与运动应激中的某些调节。
Objective:UrotensinⅡ,originally isolated from the teleost urophysis, has been reported as the most potent known vasoconstrictor.It was recently identified as an endogenous ligand for G-protein-coupled receptor 14(GPR14).Diseases closely related with insulin resistance had great impact on the human health and made more people living unhappily. More attention had been paid on aerobic exercise for it improved the insulin resistance effectively.Much more had to be studied to elucidate the exact mechanism of it.To provide more theoreticalbasis for exercise advices of insulin resistance,UrotensinⅡand GPR14 in heart,aorta and skeletal muscle of both normal and insulin resistancerats were observed under different exercise time.
Methods:To establish the rat model of insulin resistance,high fat feeding were given to 60 SD rats.Normal rats were divided into four groups randomly:normal control group(N0),normal rats swimming 1 hour a day (Nl),normal rats swimming 2 hours a day(N2)and normal rats with exhausted swimming(N3),10 rats in each group.High fat feeding rats were also divided into four groups randomly:high fat feeding rats without exercise(G0),high fat feeding rats swimming an hour a day(G1),high fat feeding rats swimming 2 hours a day(G2),high fat feeding with exhausted swimming(G3).15 rats were in each group.TG,TC,HDL,LDL and glucose in serum were examined by auto-biochemistry analysis machine. UrotensinlⅡand insulin in plasma were detected by radioimmunoassay. Immunohistochemistry technique was applied to observe the content of UrotensinⅡand GPR14 in heart,aorta and skeletal muscle.The expression of UrotensinⅡmRNA and GPR14 mRNA in these tissues were detected by Real-time PCR.
Results:Compared with normal rats,TG,TC,LDL,fasting blood glucose and insulin were all increased,while Insulin sensitive index was declined in high fat feeding group.UrotensinⅡin plasma were declined after aerobic swimming exercise,but declined after exhausted swimming in both normal and insulin resistance group.The level of UrotensinⅡand GPR14 in heart,aorta and skeletal muscle of normal rats were declined after aerobic swimming exercise but no significance was observed.The level of UrotensinⅡ,GPR14,UrotensinⅡmRNA,GPR14mRNA in heart,aorta and skeletal muscle of insulin resistance rats were declined after aerobic swimming exercise,significance was also observed.The level of UrotensinⅡand GPR14 in heart,aorta and skeletal muscle was increased after exhausted swimming.
Conclusions:Successful insulin resistance rats model were established by feeding high fat feedings.Exercise may take part in the body regulating by changing the UrotensinⅡin plasma.In both normal and insulin resistance rats,changes of UrotensinⅡand GPR14 induced by aerobic exercise were mainly in translation level or protein aero-structure,not in transcription level.The changes of UrotensinⅡand GPR14 indicated that UrotensinⅡbinding with GPR14 played a role by autocrine and paracrine in tissues.UrotensinⅡalso took part in the regulation of exercise stress in normal and insulin resistance rats.
方法:采用高脂饲料喂养方法建立胰岛素抵抗大鼠模型。随机将正常大鼠分为安静正常对照组(N0),正常+游泳1小时组(N1);正常+游泳2小时组(N2);正常+游泳力竭组(N3);每组10只。高脂膳食致胰岛素抵抗组大鼠随机分为四组:高脂膳食安静组(G0);高脂膳食+游泳1小时组(G1);高脂膳食+游泳2小时组(G2);高脂膳食+游泳力竭组(G3)每组15只。采用全自动生化分析仪测试血脂、血糖;放射免疫法测试胰岛素、血浆尾加压素Ⅱ;免疫组织化学半定量法测试心肌、主动脉、骨骼肌尾加压素Ⅱ及其受体含量;实时荧光定量PCR法测定各种组织中尾加压素Ⅱ及其受体mRNA表达。
结果:高脂膳食组大鼠血脂、空腹血糖、胰岛素均明显升高而胰岛素敏感指数下降。长时间游泳运动能够降低正常大鼠和胰岛素抵抗大鼠血浆UⅡ水平,一次性力竭游泳运动能够提高血浆UⅡ水平。长时间游泳运动能够降低正常大鼠心肌、主动脉、骨骼肌UⅡ及GPR14含量,但是它对UⅡmRNA及GPR14mRNA无显著性影响。长时间游泳运动能够降低胰岛素抵抗大鼠心肌、主动脉、骨骼肌UⅡ及GPR14含量,而且对各个组织局部UⅡmRNA及GPR14mRNA产生显著影响。在正常大鼠和胰岛素抵抗大鼠,一次性力竭运动都能够升高血浆中UⅡ水平,而且各组织UⅡ和其受体的含量均有升高趋势。
结论:本高脂饲料配方建立胰岛素抵抗大鼠模型是可行的。运动可以通过影响血浆中UⅡ来参与机体调节。有氧运动通过UⅡ及GPR14参与对正常及胰岛素抵抗大鼠局部组织的调节可能主要发生在翻译水平或者蛋白质的结构改变环节而不是在转录水平。UⅡ和GPR14在心肌、主动脉、骨骼肌分布变化的同步性提示UⅡ主要通过与其受体GPR14结合,在局部通过自分泌或者旁分泌途径发挥作用。UⅡ也参与运动应激中的某些调节。
Objective:UrotensinⅡ,originally isolated from the teleost urophysis, has been reported as the most potent known vasoconstrictor.It was recently identified as an endogenous ligand for G-protein-coupled receptor 14(GPR14).Diseases closely related with insulin resistance had great impact on the human health and made more people living unhappily. More attention had been paid on aerobic exercise for it improved the insulin resistance effectively.Much more had to be studied to elucidate the exact mechanism of it.To provide more theoreticalbasis for exercise advices of insulin resistance,UrotensinⅡand GPR14 in heart,aorta and skeletal muscle of both normal and insulin resistancerats were observed under different exercise time.
Methods:To establish the rat model of insulin resistance,high fat feeding were given to 60 SD rats.Normal rats were divided into four groups randomly:normal control group(N0),normal rats swimming 1 hour a day (Nl),normal rats swimming 2 hours a day(N2)and normal rats with exhausted swimming(N3),10 rats in each group.High fat feeding rats were also divided into four groups randomly:high fat feeding rats without exercise(G0),high fat feeding rats swimming an hour a day(G1),high fat feeding rats swimming 2 hours a day(G2),high fat feeding with exhausted swimming(G3).15 rats were in each group.TG,TC,HDL,LDL and glucose in serum were examined by auto-biochemistry analysis machine. UrotensinlⅡand insulin in plasma were detected by radioimmunoassay. Immunohistochemistry technique was applied to observe the content of UrotensinⅡand GPR14 in heart,aorta and skeletal muscle.The expression of UrotensinⅡmRNA and GPR14 mRNA in these tissues were detected by Real-time PCR.
Results:Compared with normal rats,TG,TC,LDL,fasting blood glucose and insulin were all increased,while Insulin sensitive index was declined in high fat feeding group.UrotensinⅡin plasma were declined after aerobic swimming exercise,but declined after exhausted swimming in both normal and insulin resistance group.The level of UrotensinⅡand GPR14 in heart,aorta and skeletal muscle of normal rats were declined after aerobic swimming exercise but no significance was observed.The level of UrotensinⅡ,GPR14,UrotensinⅡmRNA,GPR14mRNA in heart,aorta and skeletal muscle of insulin resistance rats were declined after aerobic swimming exercise,significance was also observed.The level of UrotensinⅡand GPR14 in heart,aorta and skeletal muscle was increased after exhausted swimming.
Conclusions:Successful insulin resistance rats model were established by feeding high fat feedings.Exercise may take part in the body regulating by changing the UrotensinⅡin plasma.In both normal and insulin resistance rats,changes of UrotensinⅡand GPR14 induced by aerobic exercise were mainly in translation level or protein aero-structure,not in transcription level.The changes of UrotensinⅡand GPR14 indicated that UrotensinⅡbinding with GPR14 played a role by autocrine and paracrine in tissues.UrotensinⅡalso took part in the regulation of exercise stress in normal and insulin resistance rats.
引文
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