不同运动方式对肥胖大鼠骨骼肌PGC-1α/FNDC5/PPARγ信号通路的影响
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摘要
目的:通过高脂膳食诱导肥胖大鼠模型,观察抗阻运动、有氧运动以及抗阻运动与有氧运动相结合3种运动方式对肥胖大鼠骨骼肌PGC-1α/FNDC5/PPARγ信号通路的影响,以及该通路在运动调控体重中的作用机制。方法:8周龄雄性SD大鼠适应性喂养1周后,随机分为正常对照组(CON组,8只)和肥胖造模组,肥胖组造模成功后,从中随机选取32只分为肥胖对照组(OC组)、肥胖有氧运动组(OAE组)、肥胖抗阻运动组(ORE组)、肥胖抗阻运动+有氧运动组(OARE组),每组8只。有氧运动采用跑台方式进行,最大强度20m/min。抗阻运动采用负重爬梯方式,适应性训练1周后,以大鼠体重的20%为初始负重,逐渐递增,最大负重量达大鼠体重的100%,隔天训练,总训练时间为12周。OARE组采用跑台和爬梯交替进行,强度与OAE组和ORE组相同。采用实时荧光定量PCR测定骨骼肌PGC-1α、FNDC5及PPARγ的mR NA相对表达量;WesternBlot法测定骨骼肌PGC-1α、FNDC5以及PPARγ蛋白的表达量。结果:1)经过12周的运动干预后,肥胖抗阻运动组、肥胖有氧运动组、肥胖抗阻运动+有氧运动组大鼠的体重都显著低于肥胖对照组(P<0.05),并且肥胖有氧运动组和肥胖抗阻运动+有氧运动组大鼠的体重显著低于肥胖抗阻运动组(P<0.05);2)肥胖有氧运动组大鼠的体脂率显著低于肥胖对照组(P<0.05),肥胖抗阻运动组以及肥胖抗阻运动+有氧运动组大鼠的体脂率低于肥胖对照组,高于肥胖有氧运动组,但都没有显著性差异(P>0.05);3)肥胖抗阻运动组、肥胖有氧运动组以及肥胖抗阻运动+有氧运动组骨骼肌PGC-1α、FNDC5、PPARγ的mR NA和蛋白表达量都显著高于肥胖对照组(P<0.05);肥胖有氧运动组骨骼肌PGC-1α、FNDC5、PPARγ的mR NA和蛋白表达量显著高于肥胖抗阻运动组以及肥胖抗阻运动+有氧运动组(P<0.05),肥胖抗阻运动+有氧运动组骨骼肌PGC-1α、FNDC5、PPARγ的mR NA和蛋白表达量显著高于肥胖抗阻运动组(P<0.05);4)大鼠骨骼肌PGC-1α、FNDC5及PPARγ的mR NA及蛋白表达量与大鼠体重、体脂重量、体脂百分比呈负相关,且都具有显著性差异(P<0.05,P<0.01)。结论:抗阻运动、有氧运动以及抗阻运动与有氧运动相结合虽然运动方式不同,但均能上调肥胖大鼠骨骼肌PGC-1α、FNDC5、PPARγ的mR NA和蛋白表达量,促进白色脂肪"棕色化",进而达到减脂的目的。研究所采用的抗阻运动以及抗阻运动与有氧运动相结合方式对肥胖大鼠骨骼肌PGC-1α/FNDC5/PPARγ信号通路的上调程度及减脂效果,虽然没有有氧运动效果好,但从时间效益上来说抗阻运动以及抗阻运动与有氧运动相结合方式所用时间短,效果也相当明显,运动强度等参数可能是导致这一现象的关键因素。
Objective: To explore the effects of resistance exercise, aerobic exercise or resistance+aerobic exercise on PGC-1α/FNDC5/PPARγ signaling pathway in skeletal muscle of obese rats and its mechanisms in exercise regulating body weight. Methods: Diet-induced obese SD rats were randomly divided into obesity control group(OC group), obesity aerobic exercise group(OAE group), obesity resistance exercise group(ORE group) and obesity resistance+aerobic exercise group(OARE group), 8 rats in each group. The aerobic exercise was performed by running on an animal treadmill, the maximum speed was 20 m/min. The resistance exercise was performed by using a load climbing ladder, after one-week adaptive training, the initial load was set as 20% of the rats' weight, the load was gradually increased to the maximum of 100% of the rats' weight. The resistance exercise was trained every two days for 12 weeks. In OARE group, the aerobic and resistance exercise were conducted alternately, and the intensity was the same as those in OAE and ORE groups. The mR NA and protein expression levels of PGC-1α, FNDC5 and PPARγ were detected by qRTPCR and Western Blotting. Results: 1) After exercise training, the body weight of 3 exercise groups were significantly lower than the OC group(P<0.05),and the OAE and OARE groups were significantly lower than the ORE group(P<0.05); 2) The body fat percentage of the OAE group was significantly lower than that of the OC group(P<0.05), the body fat percentage of the ORE and OARE groups were lower than that of the OC group but higher than the OAE group with no significant difference(P>0.05); 3) The relative expression levels of PGC-1α, FNDC5 and PPARγ mRNA and protein in the ORE, OAE, OARE groups were significantly higher than that of the OC group(P<0.05); the relative expression levels of PGC-1α, FNDC5 and PPARγ mRNA and protein in the OAE group were significantly higher than the ORE and OARE groups(P<0.05); the relative expression levels of PGC-1α, FNDC5, and PPARγ mR NA and protein in the ORE group were significantly higher than the OARE group(P<0.05); 4) The relative expression levels of PGC-1α, FNDC5, and PPARγ mRNA and protein in the gastrocnemius of rats were negatively correlated with the body weight, body fat weight and body fat percentage(P<0.05, P<0.01). Conclusion: The aerobic exercise, resistance exercise, or resistance+aerobic exercise was effectively in increasing the gene and protein expression levels of PGC-1α, FNDC5, and PPARγ, which promoted the "browning" of white fat tissue and reduced body fat. The effects of resistance exercise or resistance+aerobic exercise on the PGC-1α/FNDC5/PPARγ signaling pathway was not as effective as that of aerobic exercise, however, these two exercise types were revealed a better effect on reducing body fat in terms of time, and the exercise intensity may be the key factor leading to this phenomenon.
Objective: To explore the effects of resistance exercise, aerobic exercise or resistance+aerobic exercise on PGC-1α/FNDC5/PPARγ signaling pathway in skeletal muscle of obese rats and its mechanisms in exercise regulating body weight. Methods: Diet-induced obese SD rats were randomly divided into obesity control group(OC group), obesity aerobic exercise group(OAE group), obesity resistance exercise group(ORE group) and obesity resistance+aerobic exercise group(OARE group), 8 rats in each group. The aerobic exercise was performed by running on an animal treadmill, the maximum speed was 20 m/min. The resistance exercise was performed by using a load climbing ladder, after one-week adaptive training, the initial load was set as 20% of the rats' weight, the load was gradually increased to the maximum of 100% of the rats' weight. The resistance exercise was trained every two days for 12 weeks. In OARE group, the aerobic and resistance exercise were conducted alternately, and the intensity was the same as those in OAE and ORE groups. The mR NA and protein expression levels of PGC-1α, FNDC5 and PPARγ were detected by qRTPCR and Western Blotting. Results: 1) After exercise training, the body weight of 3 exercise groups were significantly lower than the OC group(P<0.05),and the OAE and OARE groups were significantly lower than the ORE group(P<0.05); 2) The body fat percentage of the OAE group was significantly lower than that of the OC group(P<0.05), the body fat percentage of the ORE and OARE groups were lower than that of the OC group but higher than the OAE group with no significant difference(P>0.05); 3) The relative expression levels of PGC-1α, FNDC5 and PPARγ mRNA and protein in the ORE, OAE, OARE groups were significantly higher than that of the OC group(P<0.05); the relative expression levels of PGC-1α, FNDC5 and PPARγ mRNA and protein in the OAE group were significantly higher than the ORE and OARE groups(P<0.05); the relative expression levels of PGC-1α, FNDC5, and PPARγ mR NA and protein in the ORE group were significantly higher than the OARE group(P<0.05); 4) The relative expression levels of PGC-1α, FNDC5, and PPARγ mRNA and protein in the gastrocnemius of rats were negatively correlated with the body weight, body fat weight and body fat percentage(P<0.05, P<0.01). Conclusion: The aerobic exercise, resistance exercise, or resistance+aerobic exercise was effectively in increasing the gene and protein expression levels of PGC-1α, FNDC5, and PPARγ, which promoted the "browning" of white fat tissue and reduced body fat. The effects of resistance exercise or resistance+aerobic exercise on the PGC-1α/FNDC5/PPARγ signaling pathway was not as effective as that of aerobic exercise, however, these two exercise types were revealed a better effect on reducing body fat in terms of time, and the exercise intensity may be the key factor leading to this phenomenon.
引文
陈佳红,崔建美,杨永清,等,2006.针刺和饮食结构调整对实验性肥胖大鼠减肥作用的效应研究[J].上海针灸杂志,25(4):32-34.
高艳敏,王光明,杨文礼,等,2017.高强度间歇训练和有氧运动对肥胖青年脂代谢及慢,性炎症的影响[J].中国运动医学杂志,(7):628-632.
李鹏飞,房国梁,杨星雅,2017.8周爬梯负重训练诱导大鼠比目鱼肌和趾长伸肌PGC1α/FNDC5/MSTN差异表达[J].中国运动医学杂志,36(3):195-200.
苏坤霞,2019.不同强度耐力运动影响高脂诱导肥胖模型小鼠血清Irisin含量、骨骼肌PGC-1α、FNDC5、PPAR8蛋白的表达[J].中国组织工程研究,23(3):427-434.
王静,苏有存,卢健,2011.负重爬梯训练对大鼠腓肠肌IGF-Ⅰ、IGFIEa、MGF和MSTN基因表达的影响[J].山东体育学院学报,27(5):55-58.
王京京,2013. 12周高强度间歇训练对青年肥胖女性腹部脂肪的影响[D].河北师范大学.
许杰,黄巧婷,谢敏豪,等,2018.不同强度运动对大鼠骨骼肌AMPK/PGC-1α信号通路的影响[J].成都体育学院学报,44(4):121-126.
朱惠莲,许月初,蒋卓勤,等,2002.丙酮酸对肥胖大鼠体重和脂肪代谢的影响[J].营养学报,2002,24(3):229-232.
朱小烽,王茹,杨钦,等,2017.强不同强度急性有氧运动对肥胖小鼠PGCla及其下游因子的调控影响[J].体育科学,37(3):44-50.
BAAR K, WENDE A R, JONES T E, et al.,2002. Adaptations of skeletal muscle to exercise:Rapid increase in the transcriptional coactivator PGC-1[J]. FASEB J, 16(14):1879-1886.
BOSTROM P, WU J, JEDRYCHOWSKI M P, et al.,2012. A PGC1-alphadependent myokine that drives brown-fat-like development of white fat and thermogenesis[J]. Nature,481(7382):463-468.
CHEN J Q, HUANG Y Y, GUSDON A M, et al.,2015. Irisin:A new molecular marker and target in metabolic disorder[J]. Lipids Health Dis,14(1):2.
CHINSOMBOON J, RUAS J, GUPTA R K, et al.,2009. The transcriptional coactivator PGC-1 alpha mediates exercise-induced angiogenesis in skeletal muscle[J]. Proc Natl Acad Sci USA, 106(50):21401-21406.
GUAN Y, BREYER M D, 2002. Targeting peroxisome proliferatoractivated receptors(PPARs)in kidney and urologic disease[J].Minerva Urol Nefrol, 54(2):65-79.
HANDSCHIN C, CHIN S, LI P, et al., 2007a. Skeletal muscle fibertype switching, exercise intolerance, and myopathy in PGC-1 alpha muscle-specific knock-out animals[J]. J Biol Chem, 282(41):30014-30021
HANDSCHIN C, SPIEGELMAN B M,2008b. The role of exercise and PGCla in inflammation and chronic disease[J]. Nature,454(7203):463-469.
KELLY D P,2012. Medicine. Irisin, light my fire[J]. Science,336(6077):42-43.
KRAEMER W J, VOLEK J S, CLARK K L, et al. 1999. Influence of exercise training on physiological and performance changes with weight loss in men[J]. Med Sci Sports Exerc,31(9):1320-1329.
LEE S, BARTON E R, SWEENEY H L, et al., 2004. Viral expression of insulin-like growth factor-1 enhances muscle hypertrophy in resistance-trained rats[J]. J Appl Physiol,96(3):1097-1104.
LU Y, LI H, SHEN S W, et al., 2016. Swimming exercise increases serum irisin level and reduces body fat mass in high-fat-diet fed Wistar rats[J]. Lipids Health Dis,15:93.
NARKAR V A, FAN W, DOWNES M, et al.,2011. Exercise and PGC-1 alpha-independent synchronization of type 1 muscle metabolism and vasculature by ERR gamma[J]. Cell Metab,13(3):283-293.
NORHEIM F, LANGLEITE T M, HJORTH M, et al.,2014. The effects of acute and chronic exercise on PGC-1 alpha,irisin and browning of subcutaneous adipose tissue in humans[J].FEBS J,281(3):739-749.
OHKAWARA K, TANAKA S, MIYACHI M, et al., 2007. A doseresponse relation between aerobic exercise and visceral fat reduction:Systematic review of clinical trials[J]. Int J Obes(Lond), 31(12):1786-1797.
ROBERTS M D, BAYLESS D S, COMPANY J M, et al., 2013.Elevated skeletal muscle irisin precursor FNDC5 mRNA in obese OLETF rats[J]. Metabolism,62(8):1052-1056.
ROPELLE E R, PAULI J R, CINTRA D E, et al., 2009. Acute exercise modulates the Foxol/PGC-1 alpha pathway in the liver of diet induced obesity rats[J]. J Physiol,587(9):2069-2076.
ROSEN E D,SARRAF P, TROY A E, et al. 1999. PPAR gamma is required for the differentiation of adipose tissue in vivo and in vitro[J].Mol Cell, 4(4):611-617.
SCHMITZ K H, JENSEN M D, KUGLER K C, et al., 2003. Strength training for obesity prevention in midlife women[J]. Int J Obes Relat Metab Disord, 27(3):326-333.
SHAW I, SHAW B S, 2006. Consequence of resistance training on body composition and coronary artery disease risk[J]. Cardiovasc J S Afr, 17(3):111-116.
SPIEGELMAN B M, KORSMEYER S J,2012. Irisin and the therapeutic benefits of exercise[J]. BMC Proceedings,6(Suppl 3):023.
STINE R, JAKOB G K, LOTTE L, et al., 2013. PGC-la is required for exercise-and exercise training-induced UCP1 up-regulation in mouse white adipose Tissue[J]. PLoS one, 8(5):e64123-.
TIMMONS J A, BAAR K, DAVIDSEN P K, et al., 2012. Is irisin a human exercise gene?[J]. Nature, 488(7413):E9-E10.
ULDRY M, YANG W, ST-PIERRE J, et al.,2006. Complementary action of the PGC-1 coactivators in mitochondrial biogenesis andbrown fat differentiation[J]. Cell Metab,3(5):333-341.
VILLARROYA F,2012. Irisin,turning up the heat[J]. Cell Metab,15(3):277-278.
WOODS S C, SEELEY R J, RUSHING P A, et al., 2003. A controlled high-fat diet induces an obese syndrome in rats[J]. J Nutr, 133(4):1081-1087.
WRANN C D, WHITE J P, SALOGIANNNIS J, et al.,2013. Exercise induces hippocampal BDNF through a PGC-la/FNDC5 Pathway[J].Cell Metabolism, 18(5):649-659.
YAN Z,2009. Exercise, PGC-1 alpha,and metabolic adaptation inskeletal muscle[J]. Appl Physiol Nutr Metab, 34(3):424-427.
YUAN H, NIU Y, LIU X, et al.,2014. Exercise increases the binding of MEF2A to the Cptlb promoter in mouse skeletal muscle[J]. Acta Physiologica, 12(4):283-292.
ZADEGAN F G, GHAEDI K, KALANTAR S M, et al.,2015. Cardiac differentiation of mouse embryonic stem cells is influenced by a PPARy/PGC-la-FNDC5 pathway during the stage of cardiac precursor cell formation[J]. Eur J Cell Biol,94(6):257-266.
高艳敏,王光明,杨文礼,等,2017.高强度间歇训练和有氧运动对肥胖青年脂代谢及慢,性炎症的影响[J].中国运动医学杂志,(7):628-632.
李鹏飞,房国梁,杨星雅,2017.8周爬梯负重训练诱导大鼠比目鱼肌和趾长伸肌PGC1α/FNDC5/MSTN差异表达[J].中国运动医学杂志,36(3):195-200.
苏坤霞,2019.不同强度耐力运动影响高脂诱导肥胖模型小鼠血清Irisin含量、骨骼肌PGC-1α、FNDC5、PPAR8蛋白的表达[J].中国组织工程研究,23(3):427-434.
王静,苏有存,卢健,2011.负重爬梯训练对大鼠腓肠肌IGF-Ⅰ、IGFIEa、MGF和MSTN基因表达的影响[J].山东体育学院学报,27(5):55-58.
王京京,2013. 12周高强度间歇训练对青年肥胖女性腹部脂肪的影响[D].河北师范大学.
许杰,黄巧婷,谢敏豪,等,2018.不同强度运动对大鼠骨骼肌AMPK/PGC-1α信号通路的影响[J].成都体育学院学报,44(4):121-126.
朱惠莲,许月初,蒋卓勤,等,2002.丙酮酸对肥胖大鼠体重和脂肪代谢的影响[J].营养学报,2002,24(3):229-232.
朱小烽,王茹,杨钦,等,2017.强不同强度急性有氧运动对肥胖小鼠PGCla及其下游因子的调控影响[J].体育科学,37(3):44-50.
BAAR K, WENDE A R, JONES T E, et al.,2002. Adaptations of skeletal muscle to exercise:Rapid increase in the transcriptional coactivator PGC-1[J]. FASEB J, 16(14):1879-1886.
BOSTROM P, WU J, JEDRYCHOWSKI M P, et al.,2012. A PGC1-alphadependent myokine that drives brown-fat-like development of white fat and thermogenesis[J]. Nature,481(7382):463-468.
CHEN J Q, HUANG Y Y, GUSDON A M, et al.,2015. Irisin:A new molecular marker and target in metabolic disorder[J]. Lipids Health Dis,14(1):2.
CHINSOMBOON J, RUAS J, GUPTA R K, et al.,2009. The transcriptional coactivator PGC-1 alpha mediates exercise-induced angiogenesis in skeletal muscle[J]. Proc Natl Acad Sci USA, 106(50):21401-21406.
GUAN Y, BREYER M D, 2002. Targeting peroxisome proliferatoractivated receptors(PPARs)in kidney and urologic disease[J].Minerva Urol Nefrol, 54(2):65-79.
HANDSCHIN C, CHIN S, LI P, et al., 2007a. Skeletal muscle fibertype switching, exercise intolerance, and myopathy in PGC-1 alpha muscle-specific knock-out animals[J]. J Biol Chem, 282(41):30014-30021
HANDSCHIN C, SPIEGELMAN B M,2008b. The role of exercise and PGCla in inflammation and chronic disease[J]. Nature,454(7203):463-469.
KELLY D P,2012. Medicine. Irisin, light my fire[J]. Science,336(6077):42-43.
KRAEMER W J, VOLEK J S, CLARK K L, et al. 1999. Influence of exercise training on physiological and performance changes with weight loss in men[J]. Med Sci Sports Exerc,31(9):1320-1329.
LEE S, BARTON E R, SWEENEY H L, et al., 2004. Viral expression of insulin-like growth factor-1 enhances muscle hypertrophy in resistance-trained rats[J]. J Appl Physiol,96(3):1097-1104.
LU Y, LI H, SHEN S W, et al., 2016. Swimming exercise increases serum irisin level and reduces body fat mass in high-fat-diet fed Wistar rats[J]. Lipids Health Dis,15:93.
NARKAR V A, FAN W, DOWNES M, et al.,2011. Exercise and PGC-1 alpha-independent synchronization of type 1 muscle metabolism and vasculature by ERR gamma[J]. Cell Metab,13(3):283-293.
NORHEIM F, LANGLEITE T M, HJORTH M, et al.,2014. The effects of acute and chronic exercise on PGC-1 alpha,irisin and browning of subcutaneous adipose tissue in humans[J].FEBS J,281(3):739-749.
OHKAWARA K, TANAKA S, MIYACHI M, et al., 2007. A doseresponse relation between aerobic exercise and visceral fat reduction:Systematic review of clinical trials[J]. Int J Obes(Lond), 31(12):1786-1797.
ROBERTS M D, BAYLESS D S, COMPANY J M, et al., 2013.Elevated skeletal muscle irisin precursor FNDC5 mRNA in obese OLETF rats[J]. Metabolism,62(8):1052-1056.
ROPELLE E R, PAULI J R, CINTRA D E, et al., 2009. Acute exercise modulates the Foxol/PGC-1 alpha pathway in the liver of diet induced obesity rats[J]. J Physiol,587(9):2069-2076.
ROSEN E D,SARRAF P, TROY A E, et al. 1999. PPAR gamma is required for the differentiation of adipose tissue in vivo and in vitro[J].Mol Cell, 4(4):611-617.
SCHMITZ K H, JENSEN M D, KUGLER K C, et al., 2003. Strength training for obesity prevention in midlife women[J]. Int J Obes Relat Metab Disord, 27(3):326-333.
SHAW I, SHAW B S, 2006. Consequence of resistance training on body composition and coronary artery disease risk[J]. Cardiovasc J S Afr, 17(3):111-116.
SPIEGELMAN B M, KORSMEYER S J,2012. Irisin and the therapeutic benefits of exercise[J]. BMC Proceedings,6(Suppl 3):023.
STINE R, JAKOB G K, LOTTE L, et al., 2013. PGC-la is required for exercise-and exercise training-induced UCP1 up-regulation in mouse white adipose Tissue[J]. PLoS one, 8(5):e64123-.
TIMMONS J A, BAAR K, DAVIDSEN P K, et al., 2012. Is irisin a human exercise gene?[J]. Nature, 488(7413):E9-E10.
ULDRY M, YANG W, ST-PIERRE J, et al.,2006. Complementary action of the PGC-1 coactivators in mitochondrial biogenesis andbrown fat differentiation[J]. Cell Metab,3(5):333-341.
VILLARROYA F,2012. Irisin,turning up the heat[J]. Cell Metab,15(3):277-278.
WOODS S C, SEELEY R J, RUSHING P A, et al., 2003. A controlled high-fat diet induces an obese syndrome in rats[J]. J Nutr, 133(4):1081-1087.
WRANN C D, WHITE J P, SALOGIANNNIS J, et al.,2013. Exercise induces hippocampal BDNF through a PGC-la/FNDC5 Pathway[J].Cell Metabolism, 18(5):649-659.
YAN Z,2009. Exercise, PGC-1 alpha,and metabolic adaptation inskeletal muscle[J]. Appl Physiol Nutr Metab, 34(3):424-427.
YUAN H, NIU Y, LIU X, et al.,2014. Exercise increases the binding of MEF2A to the Cptlb promoter in mouse skeletal muscle[J]. Acta Physiologica, 12(4):283-292.
ZADEGAN F G, GHAEDI K, KALANTAR S M, et al.,2015. Cardiac differentiation of mouse embryonic stem cells is influenced by a PPARy/PGC-la-FNDC5 pathway during the stage of cardiac precursor cell formation[J]. Eur J Cell Biol,94(6):257-266.