推拿联合跑轮训练对大鼠骨骼肌失神经后碱性成纤维细胞生长因子表达的实验探究
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摘要
目的:探讨推拿手法对骨骼肌失神经后碱性成纤维细胞生长因子表达的影响。方法:雄性SD大鼠64只行右下肢坐骨神经横向切断并即时外膜修复术后随机分为手法治疗组、模型对照组,每组各32只,手法治疗组给予推拿手法捏法联合跑轮治疗,模型对照组不做处理。分别在干预后4、8、12、16周各取8只大鼠检测腓肠肌肌湿重、肌卫星细胞计数及bFGF含量。结果:推拿手法组和模型对照组的肌湿重、卫星细胞数目及bFGF阳性表达面积均有所增加。肌湿重、卫星细胞仅在12周时有显著差异,其他时间点差异均不明显(P>0.05)。8周时,推拿手法组bFGF阳性表达面积略有所下降,但与模型对照组相比无统计学意义(P>0.05)。12、16周时bFGF阳性表达面积均迅速升高,且与模型对照组相比有统计学差异(P<0.05)。结论:推拿手法联合跑轮训练能够一定程度上缓解失神经造成的骨骼肌萎缩,可能与肌卫星细胞的分化增殖及bFGF生长因子的表达有关。
Objective:To investigate the effect of massage manipulation on the expression of basic fibroblast growth factor(bFGF) gene after denervation of skeletal muscle. Methods:64 male Sprague-Dawley rats underwent transverse sciatic nerve transection of the right lower extremity and were immediately repaired into a manual treatment group and a model control group, 32 rats in each group. The manual treatment group was given the massage method combined with the treadmill training. The control group did not process. Eight rats at 4, 8, 12, and 16 weeks after intervention were used to detect the wet weight of the gastrocnemius muscle, the number of muscle satellite cells, and the bFGF content. Results: Overall, the muscle wet weight, the number of satellite cells and the area of bFGF positive expression increased in massage manipulation group and model control group.Muscle wet weight and satellite cells were significantly different only at 12 weeks, and the differences were not significant at other time points(P>0.05). At 8 weeks, the positive expression area of b FGF in the manipulation group decreased slightly, but there was no statistical significance compared with the model control group(P>0.05). The positive expression area of bFGF increased rapidly at 12 and 16 weeks, and there was significant difference compared with the model control group(P<0.05).Conclusion:Massage manipulation combined with treadmill training can alleviate skeletal muscle atrophy caused by denervation to a certain extent, which may be related to the differentiation and proliferation of muscle satellite cells and the expression of bFGF.
Objective:To investigate the effect of massage manipulation on the expression of basic fibroblast growth factor(bFGF) gene after denervation of skeletal muscle. Methods:64 male Sprague-Dawley rats underwent transverse sciatic nerve transection of the right lower extremity and were immediately repaired into a manual treatment group and a model control group, 32 rats in each group. The manual treatment group was given the massage method combined with the treadmill training. The control group did not process. Eight rats at 4, 8, 12, and 16 weeks after intervention were used to detect the wet weight of the gastrocnemius muscle, the number of muscle satellite cells, and the bFGF content. Results: Overall, the muscle wet weight, the number of satellite cells and the area of bFGF positive expression increased in massage manipulation group and model control group.Muscle wet weight and satellite cells were significantly different only at 12 weeks, and the differences were not significant at other time points(P>0.05). At 8 weeks, the positive expression area of b FGF in the manipulation group decreased slightly, but there was no statistical significance compared with the model control group(P>0.05). The positive expression area of bFGF increased rapidly at 12 and 16 weeks, and there was significant difference compared with the model control group(P<0.05).Conclusion:Massage manipulation combined with treadmill training can alleviate skeletal muscle atrophy caused by denervation to a certain extent, which may be related to the differentiation and proliferation of muscle satellite cells and the expression of bFGF.
引文
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[11]赵娜,张玮,庞赓,等.基于TGF-β1/CTGF作用途径探讨推拿干预骨骼肌纤维化的作用机制[J].辽宁中医杂志,2016,43(12):2539-2541.
[12]张林峰,于天源,潘璠,等.推拿对坐骨神经损伤大鼠髓鞘碱性蛋白表达的影响简[J].中华中医药杂志,2016,31(8):3045-3048.
[13]梅旭晖,纪倩,姚斌彬,等.推拿对坐骨神经损伤大鼠神经生长因子及其受体p75NTR的影响[J].中华中医药杂志,2013,23(7):1994-1997.
[14]张林峰,于天源,潘璠,等.推拿对坐骨神经损伤大鼠层黏连蛋白表达的影响[J].西部中医药,2016,29(05):12-17.
[15]吴剑聪,耿楠,李小琴,等.推拿影响坐骨神经损伤大鼠行为学及神经营养素3、酪氨酸激酶受体C表达的研究[J].环球中医药,2014,7(3):177-183.
[16]Roudier E,Forn P,Perry M E,et al.Murine double minute-2 expression is required for capillary maintenance and exercise-induced angiogenesis in skeletal muscle[J].Faseb Journal Official Publication of the Federation of American Societies for Experimental Biology,2012,26(11):4530-4539.
[17]Armada-Da-Silva P A S,Pereira C,Amado S,et al.Chapter SixRole of Physical Exercise for Improving Posttraumatic Nerve Regeneration[J].International Review of Neurobiology,2013,109(109C):125-149.
[18]马书杰,严隽陶,陶然,等.推拿手法联合跑台训练促进大鼠坐骨神经再生的效果[J].中国康复理论与实践,2016,22(11):1276-1280.
[19]Carlson B M.The biology of long-term denervated skeletal muscle[J].European Journal of Translational Myology,2014,24(1):3293.
[20]Sambasivan R,Yao R,Kissenpfennig A,et al.Pax7-expressing satellite cells are indispensable for adult skeletal muscle regeneration[J].Development,2011,138(17):3647-3656.
[21]Antonio Musarò,Carosio S.Isolation and Culture of Satellite Cells from Mouse Skeletal Muscle[J].Methods in Molecular Biology,2017,1553:155.
[22]Snijders T,Wall B T,Dirks M L,et al.Muscle disuse atrophy is not accompanied by changes in skeletal muscle satellite cell content[J].Clinical Science,2014,126(8):557-66.
[23]Murphy M M,Lawson J A,Mathew S J,et al.Satellite cells,connective tissue fibroblasts and their interactions are crucial for muscle regeneration[J].Development,2011,138(17):3625-3637.
[24]Marzetti E,Lees H A,Manini T M,et al.Skeletal Muscle Apoptotic Signaling Predicts Thigh Muscle Volume and Gait Speed in Community-Dwelling Older Persons:An Exploratory Study[J].Plos One,2012,7(2):e32829.
[25]马书杰,严隽陶,陶然,等.手法联合跑台训练延缓周围神经损伤后骨骼肌萎缩的实验研究[J].康复学报,2016,26(05):37-40.
[26]李林峰,曾达,林伟,等.大鼠骨骼肌挫伤后bFGF与c-Fos阳性细胞率比值与损伤时间关系的研究[J].成都医学院学报,2018,13(4):405-408.
[27]Muthukrishnan L,Warder E,Mcneil P L.Basic fibroblast growth factor is efficiently released from a cytolsolic storage site through plasma membrane disruptions of endothelial cells[J].Journal of Cellular Physiology,1991,148(1):1-16.
[28]Olfert I M,Breen E C,Mathieu-Costello O,et al.Skeletal muscle capillarity and angiogenic mRNA levels after exercise training in normoxia and chronic hypoxia[J].Journal of Applied Physiology,2001,91(3):1176-1184.
[29]Chua K H,Aminuddin B S,Fuzina N H,et al.Basic fibroblast growth factor with human serum supplementation:enhancement of human chondrocyte proliferation and promotion of cartilage regeneration.[J].Singapore Medical Journal,2007,48(4):324.
[30]Majewski M,Heisterbach P,Jaquiéry C,et al.Improved tendon healing using bFGF,BMP-12 and TGFβ1 in a rat model.[J].European Cells&Materials,2018,13(35):318.
[31]Ferreira H,Martins A,Silva M L A D,et al.The functionalization of natural polymer-coated gold nanoparticles to carry bFGF to promote tissue regeneration[J].Journal of Materials Chemistry B,2018,6(14):11-16.
[32]Chakrabarti S,Islam J,Hazarika H,et al.Safety profile of silver sulfadiazine-bFGF loaded hydrogel for partial thickness burn wounds[J].Cutaneous&Ocular Toxicology,2018,37(3):258-266.
[33]Zittermann S I,Issekutz A C.Basic Fibroblast Growth Factor(bFGF,FGF-2)Potentiates Leukocyte Recruitment to Inflammation by Enhancing Endothelial Adhesion Molecule Expression[J].American Journal of Pathology,2006,168(3):0-846.
[2]Dedkov E I,Kostrominova T Y,Borisov A B,et al.Resistance vessel remodeling and reparative angiogenesis in the microcirculatory bed of long-term denervated skeletal muscles[J].Microvascular Research,2002,63(1):96-114.
[3]陆永嘉,严隽陶,马书杰,等.推拿手法联合跑轮训练对坐骨神经延期缝合大鼠神经再生的效果[J].中国康复理论与实践,2017,23(11):1273-1277.
[4]马颖,严隽陶,马书杰,等.推拿联合跑轮训练对大鼠失神经骨骼肌萎缩的效果[J].中国康复理论与实践,2018,24(5):40-44.
[5]Tandrup T,Woolf C J,Coggeshall R E.Delayed loss of small dorsal root ganglion cells after transection of the rat sciatic nerve[J].Journal of Comparative Neurology,2015,422(2):172-180.
[6]董进,柯棠山,刘坤祥.大鼠失神经不同时相的骨骼肌形态和MyoD表达变化[J].现代医药卫生,2015,31(8):1124-1126.
[7]Carlson B M.The biology of long-term denervated skeletal muscle[J].European Journal of Translational Myology,2014,24(1):3293.
[8]Yoshimura K,Harii K.A Regenerative Change during Muscle Adaptation to Denervation in Rats[J].Journal of Surgical Research,1999,81(2):0-146.
[9]郭汝宝,严隽陶,张喜林,等.推拿手法对骨骼肌失神经后卫星细胞增殖与IGF-I表达影响的实验研究[J].中华中医药学刊,2013,6(9):1872-1874.
[10]陆永嘉,严隽陶,马书杰,等.全程与间断推拿对延缓失神经大鼠骨骼肌萎缩的实验研究[J].上海中医药杂志,2017,51(10):97-99.
[11]赵娜,张玮,庞赓,等.基于TGF-β1/CTGF作用途径探讨推拿干预骨骼肌纤维化的作用机制[J].辽宁中医杂志,2016,43(12):2539-2541.
[12]张林峰,于天源,潘璠,等.推拿对坐骨神经损伤大鼠髓鞘碱性蛋白表达的影响简[J].中华中医药杂志,2016,31(8):3045-3048.
[13]梅旭晖,纪倩,姚斌彬,等.推拿对坐骨神经损伤大鼠神经生长因子及其受体p75NTR的影响[J].中华中医药杂志,2013,23(7):1994-1997.
[14]张林峰,于天源,潘璠,等.推拿对坐骨神经损伤大鼠层黏连蛋白表达的影响[J].西部中医药,2016,29(05):12-17.
[15]吴剑聪,耿楠,李小琴,等.推拿影响坐骨神经损伤大鼠行为学及神经营养素3、酪氨酸激酶受体C表达的研究[J].环球中医药,2014,7(3):177-183.
[16]Roudier E,Forn P,Perry M E,et al.Murine double minute-2 expression is required for capillary maintenance and exercise-induced angiogenesis in skeletal muscle[J].Faseb Journal Official Publication of the Federation of American Societies for Experimental Biology,2012,26(11):4530-4539.
[17]Armada-Da-Silva P A S,Pereira C,Amado S,et al.Chapter SixRole of Physical Exercise for Improving Posttraumatic Nerve Regeneration[J].International Review of Neurobiology,2013,109(109C):125-149.
[18]马书杰,严隽陶,陶然,等.推拿手法联合跑台训练促进大鼠坐骨神经再生的效果[J].中国康复理论与实践,2016,22(11):1276-1280.
[19]Carlson B M.The biology of long-term denervated skeletal muscle[J].European Journal of Translational Myology,2014,24(1):3293.
[20]Sambasivan R,Yao R,Kissenpfennig A,et al.Pax7-expressing satellite cells are indispensable for adult skeletal muscle regeneration[J].Development,2011,138(17):3647-3656.
[21]Antonio Musarò,Carosio S.Isolation and Culture of Satellite Cells from Mouse Skeletal Muscle[J].Methods in Molecular Biology,2017,1553:155.
[22]Snijders T,Wall B T,Dirks M L,et al.Muscle disuse atrophy is not accompanied by changes in skeletal muscle satellite cell content[J].Clinical Science,2014,126(8):557-66.
[23]Murphy M M,Lawson J A,Mathew S J,et al.Satellite cells,connective tissue fibroblasts and their interactions are crucial for muscle regeneration[J].Development,2011,138(17):3625-3637.
[24]Marzetti E,Lees H A,Manini T M,et al.Skeletal Muscle Apoptotic Signaling Predicts Thigh Muscle Volume and Gait Speed in Community-Dwelling Older Persons:An Exploratory Study[J].Plos One,2012,7(2):e32829.
[25]马书杰,严隽陶,陶然,等.手法联合跑台训练延缓周围神经损伤后骨骼肌萎缩的实验研究[J].康复学报,2016,26(05):37-40.
[26]李林峰,曾达,林伟,等.大鼠骨骼肌挫伤后bFGF与c-Fos阳性细胞率比值与损伤时间关系的研究[J].成都医学院学报,2018,13(4):405-408.
[27]Muthukrishnan L,Warder E,Mcneil P L.Basic fibroblast growth factor is efficiently released from a cytolsolic storage site through plasma membrane disruptions of endothelial cells[J].Journal of Cellular Physiology,1991,148(1):1-16.
[28]Olfert I M,Breen E C,Mathieu-Costello O,et al.Skeletal muscle capillarity and angiogenic mRNA levels after exercise training in normoxia and chronic hypoxia[J].Journal of Applied Physiology,2001,91(3):1176-1184.
[29]Chua K H,Aminuddin B S,Fuzina N H,et al.Basic fibroblast growth factor with human serum supplementation:enhancement of human chondrocyte proliferation and promotion of cartilage regeneration.[J].Singapore Medical Journal,2007,48(4):324.
[30]Majewski M,Heisterbach P,Jaquiéry C,et al.Improved tendon healing using bFGF,BMP-12 and TGFβ1 in a rat model.[J].European Cells&Materials,2018,13(35):318.
[31]Ferreira H,Martins A,Silva M L A D,et al.The functionalization of natural polymer-coated gold nanoparticles to carry bFGF to promote tissue regeneration[J].Journal of Materials Chemistry B,2018,6(14):11-16.
[32]Chakrabarti S,Islam J,Hazarika H,et al.Safety profile of silver sulfadiazine-bFGF loaded hydrogel for partial thickness burn wounds[J].Cutaneous&Ocular Toxicology,2018,37(3):258-266.
[33]Zittermann S I,Issekutz A C.Basic Fibroblast Growth Factor(bFGF,FGF-2)Potentiates Leukocyte Recruitment to Inflammation by Enhancing Endothelial Adhesion Molecule Expression[J].American Journal of Pathology,2006,168(3):0-846.