目的:探究不同强度一次性运动干预对大鼠睾周白色脂肪中FNDC5和UCP-1表达量的影响。方法:30只SD大鼠随机分为对照组(C组,n=6)和运动组(E组,n=24)。E组大鼠随机分为中等强度运动组(EM组,n=12,15 m/min)和高强度间歇运动组(EH组,n=12,35 m...目的:探究不同强度一次性运动干预对大鼠睾周白色脂肪中FNDC5和UCP-1表达量的影响。方法:30只SD大鼠随机分为对照组(C组,n=6)和运动组(E组,n=24)。E组大鼠随机分为中等强度运动组(EM组,n=12,15 m/min)和高强度间歇运动组(EH组,n=12,35 m/min,6 min间歇5 min,重复3次),分别在一次性运动后即刻和6 h后取大鼠睾周白色脂肪,利用RT-q PCR法检测睾周白色脂肪组织中细胞外跨膜受体Ⅲ型纤连蛋白域蛋白5(FNDC5)和解偶联蛋白1(UCP-1)m RNA表达水平,Western Blot法检测睾周白色脂肪组织中FNDC5和UCP-1蛋白含量。结果:(1)与C组相比,EH组运动后即刻FNDC5 m RNA表达水平显著升高(P<0.01)。(2)与C组相比,运动组大鼠睾周白色脂肪组织中UCP-1m RNA水平在运动后即刻显著降低(P<0.05);与运动后即刻相比较,运动后6 h显著升高(P<0.05)。(3)与C组相比,EM和EH组FNDC5蛋白含量均有上升趋势,其中EM组显著升高(P<0.05)。(4)与C组相比,EM组和EH组UCP-1蛋白含量均有上升趋势,其中EH组运动后即刻出现显著升高(P<0.05)。结论:(1)一次性运动能够提高大鼠睾周白色脂肪FNDC5和UCP-1蛋白表达水平,促进白色脂肪棕色化,增加脂肪组织产热;(2)一次性运动后FNDC5 m RNA和UCP-1 m RNA的变化具有时效性,其中UCP-1 m RNA在运动后6小时出现显著升高,而FNDC5 m RNA在高强度间歇运动后即刻变化最明显。展开更多
Purpose： Fibronectin type III domain-containing protein 5 （FNDC5）, also known as irisin, is a myokine secreted from muscle in response to exercise and improves obesity and glucose homeostasis. However, the molecula...Purpose： Fibronectin type III domain-containing protein 5 （FNDC5）, also known as irisin, is a myokine secreted from muscle in response to exercise and improves obesity and glucose homeostasis. However, the molecular mecha- nisms that regulate FNDC5 expression and the functional significance of FNDC5 in skeletal muscle remain un- known. In this study, we explored the possible pathways that induce FNDC5 expression and delineated its metabol- ic effects on skeletal muscle. Methods： C2C12 myotubes were treated with various concentrations of Sp-cAMP, forskolin, and ionomycin respectively for various durations. FNDC5 and related metabolic genes＇ expressions were measured by quantitative real-time polymerase chain reaction （qRT-PCR）. Cyclic AMP responsive element-binding protein （CREB） phosphorylation was measured by Western blot. Oxidative phosphorylation was quantified by oxy- gen consumption rate （OCR） measurement using XF-96 analyzer （Seahorse Bioscience）. The statistical signifi- cance was calculated by one-way analysis of variance （ANOVA）. Data were considered significant when P 〈 0.05. Results： We found that cAMP and forskolin dose and time dependently increased FNDC5 expression in C2C12 myotubes. A synergistic effect of forskolin and ionomycin on FNDC5 expression was also found. CREB phosphoryl- ation was elevated in myotubes simultaneously upon these treatments. C2C12 myotubes over expressing CREB dis- plays increased FNDC5 expression as well, suggesting CREB was a regulator of FNDC5 expression. Functionally, irisin treatment enhanced mitochondrial biogenesis of C2C12 myotubes through increasing peroxisome proliferator- activated receptor gamma coactivator-1α （PGC-1α）, nuclear respiratory factor 1 （NRF1） and mitochondrial tran-scription factor A （TFAM） expressions, leading to increase myotube mitochondrial respirations and ATP produc- tion. Conclusions Our observation indicates that irisin is a metabolic modulator of skeletal muscle, whose expres- sion is controlled by cAMP pathway and intracellular level of calcium.展开更多
Irisin为一种新发现的肌肉因子,是蛋白水解酶剪切Ⅲ型纤连蛋白组件包含蛋白5(fibrone-ctin typeⅢ domain-containing protein 5,FNDC5)后形成的可分泌多肽片段。运动能上调骨骼肌FNDC5的mRNA表达和循环中irisin水平。Irisin能促进脂肪...Irisin为一种新发现的肌肉因子,是蛋白水解酶剪切Ⅲ型纤连蛋白组件包含蛋白5(fibrone-ctin typeⅢ domain-containing protein 5,FNDC5)后形成的可分泌多肽片段。运动能上调骨骼肌FNDC5的mRNA表达和循环中irisin水平。Irisin能促进脂肪细胞解耦联蛋白(uncoupling protein1,UCP1)表达、诱导脂肪小滴形成、增加线粒体密度、增加脂肪细胞氧耗、导致脂肪细胞向褐色脂肪细胞表型转变,从而抑制肥胖和胰岛素抵抗的发生,是代谢性疾病防治的新靶点。展开更多
[目的]探究Ⅲ型纤连蛋白域包含蛋白5(typeⅢfibronectin domain-containing protein 5,FNDC5)基因对藏猪低氧适应性的调控作用。[方法]以藏猪和大约克猪为实验动物,提取其耳组织DNA,分段扩增FNDC5基因的5′侧翼区、3′侧翼区和CDS区,测...[目的]探究Ⅲ型纤连蛋白域包含蛋白5(typeⅢfibronectin domain-containing protein 5,FNDC5)基因对藏猪低氧适应性的调控作用。[方法]以藏猪和大约克猪为实验动物,提取其耳组织DNA,分段扩增FNDC5基因的5′侧翼区、3′侧翼区和CDS区,测序后筛选单核苷酸多态性(SNPs)位点。采集实验猪心脏、肺脏组织,提取其RNA及总蛋白,采用实时荧光定量PCR(RT-qPCR)法检测FNDC5基因mRNA的表达量,采用Western blot法检测FNDC5蛋白的表达量。采集藏猪和大约克猪颈动脉血样各30头份,测定血液生理指标。将猪骨骼肌卫星细胞(PSMSC)分为常氧组(氧气体积分数21%)及体积分数5%和1%氧气组,常氧组又分为PGC-1α激活剂ZLN005添加组、PGC-1α抑制剂SR-18292添加组和空白对照组,培养后分别采用RT-qPCR和Western blot法检测FNDC5基因mRNA和蛋白的表达量。[结果]在FNDC5基因CDS区,2种猪未检测到SNPs位点;在5′侧翼区,2种猪有3个SNPs位点,分别为g.-118A>G、g.-790T>C和g.-931A>G;在3′侧翼区,大约克猪无SNP位点,藏猪有4个多态性位点(分别为g.46G>T、g.56G>C、g.517T>C和g.1028G>A)。这些SNPs可能是藏猪低氧适应性的重要调控位点。藏猪的白细胞数、红细胞数、血红蛋白质量浓度、红细胞压积、血小板计数、血小板压积指标极显著高于大约克猪(P<0.01),红细胞分布宽度变异系数极显著低于大约克猪(P<0.01),其他指标与大约克猪无显著差异,推测藏猪具有更强的低氧适应能力。藏猪心脏FNDC5基因和蛋白表达量极显著高于大约克猪,肺脏FNDC5基因表达量极显著高于大约克猪。在PSMSC中,FNDC5基因和蛋白的表达量均随着氧体积分数的降低而显著或极显著降低;PGC-1α激活剂ZLN005可极显著促进FNDC5基因的表达,抑制剂SR-18292可极显著抑制FNDC5基因的表达。[结论]FNDC5基因和部分血液生理指标可能与藏猪的低氧适应性相关;PGC-1α可正向调控FNDC5基因的表达。展开更多
目的:通过高脂膳食诱导肥胖小鼠模型,观察不同强度急性运动对肥胖小鼠PGC-1α及其下游因子的转录与翻译水平的影响,进一步探讨PGC-1α信号在运动减控体重中的生理机制。方法:C57BL/6J雄性小鼠经高脂膳食诱导建模成功后,随机分为安静对照...目的:通过高脂膳食诱导肥胖小鼠模型,观察不同强度急性运动对肥胖小鼠PGC-1α及其下游因子的转录与翻译水平的影响,进一步探讨PGC-1α信号在运动减控体重中的生理机制。方法:C57BL/6J雄性小鼠经高脂膳食诱导建模成功后,随机分为安静对照组(C组)、小强度(SE组)、中强度(ME组)、大强度有氧运动组(HE组)。后3组进行不同强度的一次性跑台训练,运动后即刻处死。采用ELISA测定血清TNF-α、IRISIN、MCAD、CPT-1含量;实时荧光定量PCR测定骨骼肌PGC-1α、PPARγ、PPARβ、TNF-α及FNDC5 m RNA相对表达量。结果:与C组比较,SE组小鼠血清MCAD显著性提升(P<0.05);ME和HE组TNF-α显著性下降(P<0.05;P<0.01);CPT-1与Irisin4组间差异不具有显著性(P>0.05);与C组相比,SE、ME及HE组骨骼肌PGC-1αm RNA相对表达量显著性提升(P<0.01);ME与HE组PPARγm RNA显著性增加(P<0.05;P<0.01);ME与HE组PPARβm RNA显著性增加(P<0.01;P<0.05);3个运动组中TNF-αm RNA相对表达量显著性降低(P<0.01);ME与HE组FNDC5 m RNA相对表达量显著性增加(P<0.05)。骨骼肌PGC-1αm RNA与FNDC5 m RNA相对表达量呈显著性正相关(P<0.01);骨骼肌FNDC5 m RNA相对表达量与血清Irisin含量相关性较低(P>0.05);骨骼肌PGC-1αm RNA相对表达量与血清TNF-α含量显著性正相关(P<0.05)。结论:急性有氧运动可以显著增强PGC-1α及其下游因子的转录与翻译水平;小鼠骨骼肌氧化代谢和炎症反应之间可能存在相互拮抗的肌肉分子机制。展开更多
文摘目的:探究不同强度一次性运动干预对大鼠睾周白色脂肪中FNDC5和UCP-1表达量的影响。方法:30只SD大鼠随机分为对照组(C组,n=6)和运动组(E组,n=24)。E组大鼠随机分为中等强度运动组(EM组,n=12,15 m/min)和高强度间歇运动组(EH组,n=12,35 m/min,6 min间歇5 min,重复3次),分别在一次性运动后即刻和6 h后取大鼠睾周白色脂肪,利用RT-q PCR法检测睾周白色脂肪组织中细胞外跨膜受体Ⅲ型纤连蛋白域蛋白5(FNDC5)和解偶联蛋白1(UCP-1)m RNA表达水平,Western Blot法检测睾周白色脂肪组织中FNDC5和UCP-1蛋白含量。结果:(1)与C组相比,EH组运动后即刻FNDC5 m RNA表达水平显著升高(P<0.01)。(2)与C组相比,运动组大鼠睾周白色脂肪组织中UCP-1m RNA水平在运动后即刻显著降低(P<0.05);与运动后即刻相比较,运动后6 h显著升高(P<0.05)。(3)与C组相比,EM和EH组FNDC5蛋白含量均有上升趋势,其中EM组显著升高(P<0.05)。(4)与C组相比,EM组和EH组UCP-1蛋白含量均有上升趋势,其中EH组运动后即刻出现显著升高(P<0.05)。结论:(1)一次性运动能够提高大鼠睾周白色脂肪FNDC5和UCP-1蛋白表达水平,促进白色脂肪棕色化,增加脂肪组织产热;(2)一次性运动后FNDC5 m RNA和UCP-1 m RNA的变化具有时效性,其中UCP-1 m RNA在运动后6小时出现显著升高,而FNDC5 m RNA在高强度间歇运动后即刻变化最明显。
文摘Purpose： Fibronectin type III domain-containing protein 5 （FNDC5）, also known as irisin, is a myokine secreted from muscle in response to exercise and improves obesity and glucose homeostasis. However, the molecular mecha- nisms that regulate FNDC5 expression and the functional significance of FNDC5 in skeletal muscle remain un- known. In this study, we explored the possible pathways that induce FNDC5 expression and delineated its metabol- ic effects on skeletal muscle. Methods： C2C12 myotubes were treated with various concentrations of Sp-cAMP, forskolin, and ionomycin respectively for various durations. FNDC5 and related metabolic genes＇ expressions were measured by quantitative real-time polymerase chain reaction （qRT-PCR）. Cyclic AMP responsive element-binding protein （CREB） phosphorylation was measured by Western blot. Oxidative phosphorylation was quantified by oxy- gen consumption rate （OCR） measurement using XF-96 analyzer （Seahorse Bioscience）. The statistical signifi- cance was calculated by one-way analysis of variance （ANOVA）. Data were considered significant when P 〈 0.05. Results： We found that cAMP and forskolin dose and time dependently increased FNDC5 expression in C2C12 myotubes. A synergistic effect of forskolin and ionomycin on FNDC5 expression was also found. CREB phosphoryl- ation was elevated in myotubes simultaneously upon these treatments. C2C12 myotubes over expressing CREB dis- plays increased FNDC5 expression as well, suggesting CREB was a regulator of FNDC5 expression. Functionally, irisin treatment enhanced mitochondrial biogenesis of C2C12 myotubes through increasing peroxisome proliferator- activated receptor gamma coactivator-1α （PGC-1α）, nuclear respiratory factor 1 （NRF1） and mitochondrial tran-scription factor A （TFAM） expressions, leading to increase myotube mitochondrial respirations and ATP produc- tion. Conclusions Our observation indicates that irisin is a metabolic modulator of skeletal muscle, whose expres- sion is controlled by cAMP pathway and intracellular level of calcium.
文摘目的:通过高脂膳食诱导肥胖小鼠模型,观察不同强度急性运动对肥胖小鼠PGC-1α及其下游因子的转录与翻译水平的影响,进一步探讨PGC-1α信号在运动减控体重中的生理机制。方法:C57BL/6J雄性小鼠经高脂膳食诱导建模成功后,随机分为安静对照组(C组)、小强度(SE组)、中强度(ME组)、大强度有氧运动组(HE组)。后3组进行不同强度的一次性跑台训练,运动后即刻处死。采用ELISA测定血清TNF-α、IRISIN、MCAD、CPT-1含量;实时荧光定量PCR测定骨骼肌PGC-1α、PPARγ、PPARβ、TNF-α及FNDC5 m RNA相对表达量。结果:与C组比较,SE组小鼠血清MCAD显著性提升(P<0.05);ME和HE组TNF-α显著性下降(P<0.05;P<0.01);CPT-1与Irisin4组间差异不具有显著性(P>0.05);与C组相比,SE、ME及HE组骨骼肌PGC-1αm RNA相对表达量显著性提升(P<0.01);ME与HE组PPARγm RNA显著性增加(P<0.05;P<0.01);ME与HE组PPARβm RNA显著性增加(P<0.01;P<0.05);3个运动组中TNF-αm RNA相对表达量显著性降低(P<0.01);ME与HE组FNDC5 m RNA相对表达量显著性增加(P<0.05)。骨骼肌PGC-1αm RNA与FNDC5 m RNA相对表达量呈显著性正相关(P<0.01);骨骼肌FNDC5 m RNA相对表达量与血清Irisin含量相关性较低(P>0.05);骨骼肌PGC-1αm RNA相对表达量与血清TNF-α含量显著性正相关(P<0.05)。结论:急性有氧运动可以显著增强PGC-1α及其下游因子的转录与翻译水平;小鼠骨骼肌氧化代谢和炎症反应之间可能存在相互拮抗的肌肉分子机制。
