针刺对一次离心运动模型大鼠白细胞介素6表达的影响
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摘要
背景:白细胞介素6是一种细胞因子,其来源广泛,在运动后骨骼肌能够大量产生并对运动功能恢复起到重要作用;运动后进行骨骼肌针刺能够改善骨骼肌的超微结构,降低损伤程度,促进功能恢复。目前尚未见到针刺对运动后白细胞介素6影响的报道。目的:探究针刺干预对一次离心运动后大鼠白细胞介素6的影响。方法:114只雄性SD大鼠,随机分为:安静对照组(n=6)、单纯运动组(n=36)、单纯针刺组(n=36)、运动针刺组(n=36)。单纯运动组和运动针刺组均在坡度-16°的跑台上进行一次离心运动,单纯针刺组和运动针刺组进行小腿三头肌的斜刺。各组处理后,除安静组直接取材,其余3组分别在干预后0,2,24,48,72,120h时间点取材,每个时间点6只。Western Blot法分别检测比目鱼肌、股直肌白细胞介素6的蛋白表达;ELISA法检测血浆中白细胞介素6水平。结果与结论:(1)单纯针刺处理后,比目鱼肌和股直肌中白细胞介素6蛋白表达以及血浆中白细胞介素6水平均无明显变化(P> 0.05);(2)一次离心运动后,比目鱼肌、股直肌和血浆中白细胞介素6均出现不同程度升高,且比目鱼肌与股直肌间白细胞介素6表达无明显差异(P> 0.05);(3)运动后针刺能明显下调比目鱼肌白细胞介素6蛋白峰值表达(P <0.05),而股直肌和血浆中白细胞介素6表达无明显变化(P> 0.05);(4)结果表明,对骨骼肌进行直接针刺干预不能引起白细胞介素6明显变化,骨骼肌在离心运动后会分泌较多的白细胞介素6,同时进行针刺干预能够降低针刺部位白细胞介素6的峰值量;针刺能够在局部对白细胞介素6产生明显作用,对于其他部位的骨骼肌和血浆未能产生显著效果。
BACKGROUND: Interleukin-6,is a cytokine that has wide sources. Interleukin-6 increases in the skeletal muscle after exercise,which plays an important role in motor function recovery. Acupuncture at the skeletal muscle after exercise can improve the ultrastructure of skeletal mscle,alleviate injury and promote functional recovery. However,acupuncture effects on the interleukin-6 post exercise remains unknown. OBJECTIVE: To explore the influence of acupuncture intervention on interleukin-6 in rats after eccentric exercise. METHODS: Totally 114 male Sprague-Dawley rats were randomly divided into four groups: control group(n=6),exercise group(n=36),acupuncture group(n=36) and exercise plus acupuncture group(n=36). The rats in the exercise and exercise plus acupuncture groups were submitted to eccentric exercise on a downward treadmill at 16° decline. The rats in the acupuncture and exercise plus acupuncture groups were subjected to the oblique acupuncture at the triceps surae. Samples except the control group were collected at 0,2,24,48,72 and 120 hours after treatment(n=6 per group at each time point). Protein expression level of interleukin-6 in the soleus and rectus femoris was detected by western blot assay. The plasma level of interleukin-6 was determined by ELISA. RESULTS AND CONCLUSION: In the acupuncture group,the protein expression level of interleukin-6 in the soleus and rectus femoris and plasma level of interleukin-6 showed no significant changes(P > 0.05). After eccentric exercise,the interleukin-6 levels in the soleus,rectus femoris and plasma were significantly increased,and the protein expression level of interleukin-6 did not differ significantly between soleus and rectus femoris(P > 0.05). Acupuncture intervention after exercise,the protein expression level of interleukin-6 in the soleus was significantly reduced(P < 0.05),while interleukin-6 levels in the rectus femoris and plasma showed no significant change(P > 0.05). These findings imply that acupuncture cannot markedly change the interleukin-6 level in normal rats. Eccentric exercise can lead skeletal muscle to create more interleukin-6,and acupuncture intervention after eccentric exercise can reduce the amount of interleukin-6 at peak phase in the needling part. But its effect is local,not significant on the skeletal muscle of other parts and plasma.
BACKGROUND: Interleukin-6,is a cytokine that has wide sources. Interleukin-6 increases in the skeletal muscle after exercise,which plays an important role in motor function recovery. Acupuncture at the skeletal muscle after exercise can improve the ultrastructure of skeletal mscle,alleviate injury and promote functional recovery. However,acupuncture effects on the interleukin-6 post exercise remains unknown. OBJECTIVE: To explore the influence of acupuncture intervention on interleukin-6 in rats after eccentric exercise. METHODS: Totally 114 male Sprague-Dawley rats were randomly divided into four groups: control group(n=6),exercise group(n=36),acupuncture group(n=36) and exercise plus acupuncture group(n=36). The rats in the exercise and exercise plus acupuncture groups were submitted to eccentric exercise on a downward treadmill at 16° decline. The rats in the acupuncture and exercise plus acupuncture groups were subjected to the oblique acupuncture at the triceps surae. Samples except the control group were collected at 0,2,24,48,72 and 120 hours after treatment(n=6 per group at each time point). Protein expression level of interleukin-6 in the soleus and rectus femoris was detected by western blot assay. The plasma level of interleukin-6 was determined by ELISA. RESULTS AND CONCLUSION: In the acupuncture group,the protein expression level of interleukin-6 in the soleus and rectus femoris and plasma level of interleukin-6 showed no significant changes(P > 0.05). After eccentric exercise,the interleukin-6 levels in the soleus,rectus femoris and plasma were significantly increased,and the protein expression level of interleukin-6 did not differ significantly between soleus and rectus femoris(P > 0.05). Acupuncture intervention after exercise,the protein expression level of interleukin-6 in the soleus was significantly reduced(P < 0.05),while interleukin-6 levels in the rectus femoris and plasma showed no significant change(P > 0.05). These findings imply that acupuncture cannot markedly change the interleukin-6 level in normal rats. Eccentric exercise can lead skeletal muscle to create more interleukin-6,and acupuncture intervention after eccentric exercise can reduce the amount of interleukin-6 at peak phase in the needling part. But its effect is local,not significant on the skeletal muscle of other parts and plasma.
引文
[1]方剑乔,邵晓梅.针刺镇痛的新思路--针灸参与疼痛多维度调节的可行性[J].针刺研究,2017,42(1):85-89.
[2]严晓艺,贾丽燕,张云皎,等.针刺治疗脑卒中后肌痉挛的系统综述与Meta分析[J].北京中医药大学学报,2017,40(1):52-58.
[3]高纺,盛红梅,张田宁,等.针刺不同穴方对便秘小鼠肠运动的影响[J].针刺研究,2017,42(1):62-66+75.
[4]尚画雨,付玉,夏志,等.针刺对运动性骨骼肌损伤大鼠骨骼肌线粒体自噬的影响[J].中国病理生理杂志,2017,33(11):2038-2046.
[5]丁海丽.运动及针刺干预对骨骼肌内质网应激-自噬反应的影响机制研究[D].北京:北京体育大学,2017.
[6]刘阳,王瑞元,康红哲.不同时程下[Ca~(2+)]介导骨骼肌微细损伤与修复的机制及针刺的干预作用[J].北京体育大学学报,2015,38(2):54-60.
[7]孙君志.离心运动诱导骨骼肌细胞微管损伤的机制及针刺干预研究[D].北京:北京体育大学,2014.
[8]白胜超,赵晓琴,孙君志,等.针刺对一次离心运动后大鼠炎症反应的影响[J].首都体育学院学报,2017,29(1):85-90.
[9]Baeza-Raja B,Mu?oz-Cánoves P.p38 MAPK-induced nuclear factor-kappa B activity is required for skeletal muscle differentiation:role of interleukin-6.Mol Biol Cell.2004;15(4):2013-2026.
[10]唐晖,谢敏豪.运动引起肌源性IL-6分泌的研究进展[J].中国运动医学杂志,2006,25(1):60-66.
[11]王瑞元,周越.骨骼肌与运动[M].北京:人民体育出版社,2013:1-19.
[12]Armstrong RB,Ogilvie RW,Schwane JA.Eccentric exercise-induced injury to rat skeletal muscle J Appl Physiol Respir Environ Exerc Physiol.1983;54(1):80-93.
[13]Mcginnis GR,Ballmann C,Peters B,et al.Interleukin-6 mediates exercise preconditioning against myocardial ischemia reperfusion injury.Am J Physiol Heart Circ Physiol.2015;308(11):H1423-1433.
[14]Ameis K,Trouth AJ,Bond V,et al.Degeneration/Regeneration as a Mechanism Contributing to the Effect of Manual AcupunctureInduced Injury on Rat Skeletal Muscle.Intech,2011.
[15]Pedersen BK,Hoffman-Goetz L.Exercise and the immune system:regulation,integration,and adaptation.Physiol Rev.2000;80(3):1055-1081.
[16]Ullum H,Haahr PM,Diamant M,et al.Bicycle exercise enhances plasma IL-6 but does not change IL-1 alpha,IL-1 beta,IL-6,or TNF-alpha pre-m RNA in BMNC.J Appl Physiol(1985).1994;77(1):93-97.
[17]Steensberg A,van Hall G,Osada T,et al.Production of interleukin-6 in contracting human skeletal muscles can account for the exercise-induced increase in plasma interleukin-6.JPhysiol.2000;529(Pt 1):237-242.
[18]苏全生,田野,王东辉,等.大蒜素对大鼠离心运动后血IL-6、CK、CK-MM和Ca2+变化的影响[J].体育科学,2006,26(3):73-76.
[19]赵广高,苏全生,张慧,等.大蒜素与联合抗氧化剂干预对运动员DOMS和血浆IL-6水平变化的影响[J].体育科学,2008,28(7):66-71.
[20]Rohde T,Mac Lean DA,Richter EA,et al.Prolonged submaximal eccentric exercise is associated with increased levels of plasma IL-6.Am J Physiol.1997;273(1 Pt 1):E85-91.
[21]Ostrowski K,Rohde T,Zacho M,et al.Evidence that interleukin-6is produced in human skeletal muscle during prolonged running.JPhysiol.1998;508(Pt 3):949-953.
[22]Bruunsgaard H,Galbo H,Halkjaer-Kristensen J,et al.Exerciseinduced increase in serum interleukin-6 in humans is related to muscle damage.J Physiol.1997;499(Pt 3):833-841.
[23]Hiscock N,Chan MH,Bisucci T,et al.Skeletal myocytes are a source of interleukin-6 m RNA expression and protein release during contraction:evidence of fiber type specificity.FASEB J.2004;18(9):992-994.
[24]Fischer CP,Hiscock NJ,Penkowa M,et al.Supplementation with vitamins C and E inhibits the release of interleukin-6 from contracting human skeletal muscle.J Physiol.2004;558(Pt 2):633-645.
[25]Pernille K,Milena P,Charlotte K,et al.Interleukin-6 receptor expression in contracting human skeletal muscle;regulating role of IL-6.FASEB J.2005;19(9):1181-1183.
[26]王今越,丁树哲,刘伟,等.运动与IL-6的研究进展[J].体育科学,2007,27(6):60-69.
[27]林玉敏.艾灸对慢性疲劳模型大鼠的免疫干预及临床研究[D].广州:广州中医药大学,2014.
[28]Hsu CP,Chung YC.Influence,of interleukin-6,on the invasiveness,of human,colorectal,carcinoma.Anticancer Research,2016,26(6B):4607-4614.
[29]Roxburgh CS,Mcmillan DC.Therapeutics targeting innate immune/inflammatory responses through the interleukin-6/JAK/STAT signal transduction pathway in patients with cancer.Transl Res.2016;167(1):61-66
[30]郑盛,杨涓,陈文钦,等.人脐带间充质干细胞通过IL-6/STAT3信号通路促进肝癌Hep G-2细胞的增殖[J].中国组织工程研究,2016,20(50):7460-7468
[2]严晓艺,贾丽燕,张云皎,等.针刺治疗脑卒中后肌痉挛的系统综述与Meta分析[J].北京中医药大学学报,2017,40(1):52-58.
[3]高纺,盛红梅,张田宁,等.针刺不同穴方对便秘小鼠肠运动的影响[J].针刺研究,2017,42(1):62-66+75.
[4]尚画雨,付玉,夏志,等.针刺对运动性骨骼肌损伤大鼠骨骼肌线粒体自噬的影响[J].中国病理生理杂志,2017,33(11):2038-2046.
[5]丁海丽.运动及针刺干预对骨骼肌内质网应激-自噬反应的影响机制研究[D].北京:北京体育大学,2017.
[6]刘阳,王瑞元,康红哲.不同时程下[Ca~(2+)]介导骨骼肌微细损伤与修复的机制及针刺的干预作用[J].北京体育大学学报,2015,38(2):54-60.
[7]孙君志.离心运动诱导骨骼肌细胞微管损伤的机制及针刺干预研究[D].北京:北京体育大学,2014.
[8]白胜超,赵晓琴,孙君志,等.针刺对一次离心运动后大鼠炎症反应的影响[J].首都体育学院学报,2017,29(1):85-90.
[9]Baeza-Raja B,Mu?oz-Cánoves P.p38 MAPK-induced nuclear factor-kappa B activity is required for skeletal muscle differentiation:role of interleukin-6.Mol Biol Cell.2004;15(4):2013-2026.
[10]唐晖,谢敏豪.运动引起肌源性IL-6分泌的研究进展[J].中国运动医学杂志,2006,25(1):60-66.
[11]王瑞元,周越.骨骼肌与运动[M].北京:人民体育出版社,2013:1-19.
[12]Armstrong RB,Ogilvie RW,Schwane JA.Eccentric exercise-induced injury to rat skeletal muscle J Appl Physiol Respir Environ Exerc Physiol.1983;54(1):80-93.
[13]Mcginnis GR,Ballmann C,Peters B,et al.Interleukin-6 mediates exercise preconditioning against myocardial ischemia reperfusion injury.Am J Physiol Heart Circ Physiol.2015;308(11):H1423-1433.
[14]Ameis K,Trouth AJ,Bond V,et al.Degeneration/Regeneration as a Mechanism Contributing to the Effect of Manual AcupunctureInduced Injury on Rat Skeletal Muscle.Intech,2011.
[15]Pedersen BK,Hoffman-Goetz L.Exercise and the immune system:regulation,integration,and adaptation.Physiol Rev.2000;80(3):1055-1081.
[16]Ullum H,Haahr PM,Diamant M,et al.Bicycle exercise enhances plasma IL-6 but does not change IL-1 alpha,IL-1 beta,IL-6,or TNF-alpha pre-m RNA in BMNC.J Appl Physiol(1985).1994;77(1):93-97.
[17]Steensberg A,van Hall G,Osada T,et al.Production of interleukin-6 in contracting human skeletal muscles can account for the exercise-induced increase in plasma interleukin-6.JPhysiol.2000;529(Pt 1):237-242.
[18]苏全生,田野,王东辉,等.大蒜素对大鼠离心运动后血IL-6、CK、CK-MM和Ca2+变化的影响[J].体育科学,2006,26(3):73-76.
[19]赵广高,苏全生,张慧,等.大蒜素与联合抗氧化剂干预对运动员DOMS和血浆IL-6水平变化的影响[J].体育科学,2008,28(7):66-71.
[20]Rohde T,Mac Lean DA,Richter EA,et al.Prolonged submaximal eccentric exercise is associated with increased levels of plasma IL-6.Am J Physiol.1997;273(1 Pt 1):E85-91.
[21]Ostrowski K,Rohde T,Zacho M,et al.Evidence that interleukin-6is produced in human skeletal muscle during prolonged running.JPhysiol.1998;508(Pt 3):949-953.
[22]Bruunsgaard H,Galbo H,Halkjaer-Kristensen J,et al.Exerciseinduced increase in serum interleukin-6 in humans is related to muscle damage.J Physiol.1997;499(Pt 3):833-841.
[23]Hiscock N,Chan MH,Bisucci T,et al.Skeletal myocytes are a source of interleukin-6 m RNA expression and protein release during contraction:evidence of fiber type specificity.FASEB J.2004;18(9):992-994.
[24]Fischer CP,Hiscock NJ,Penkowa M,et al.Supplementation with vitamins C and E inhibits the release of interleukin-6 from contracting human skeletal muscle.J Physiol.2004;558(Pt 2):633-645.
[25]Pernille K,Milena P,Charlotte K,et al.Interleukin-6 receptor expression in contracting human skeletal muscle;regulating role of IL-6.FASEB J.2005;19(9):1181-1183.
[26]王今越,丁树哲,刘伟,等.运动与IL-6的研究进展[J].体育科学,2007,27(6):60-69.
[27]林玉敏.艾灸对慢性疲劳模型大鼠的免疫干预及临床研究[D].广州:广州中医药大学,2014.
[28]Hsu CP,Chung YC.Influence,of interleukin-6,on the invasiveness,of human,colorectal,carcinoma.Anticancer Research,2016,26(6B):4607-4614.
[29]Roxburgh CS,Mcmillan DC.Therapeutics targeting innate immune/inflammatory responses through the interleukin-6/JAK/STAT signal transduction pathway in patients with cancer.Transl Res.2016;167(1):61-66
[30]郑盛,杨涓,陈文钦,等.人脐带间充质干细胞通过IL-6/STAT3信号通路促进肝癌Hep G-2细胞的增殖[J].中国组织工程研究,2016,20(50):7460-7468