Objective To explore the effect and mechanism of Tuina on denervated skeletal muscle atrophy.
Methods A total of 77 male Sprague-Dawley rats were randomly divided into sham group (n = 7), model group (n = 35) and Tuina group (n = 35). The latter two groups were established skeletal muscle atrophy model by exposing and cutting off the common tibial nerve of rats. One day after modeling, the lower limbs of the surgical side received Tuina in Tuina group. Separately, the surgical side of gastrocnemius muscle were sampled on the 0th, 7th, 14th, 21st and 28th day after modeling, and measured the wet mass ratio. The cross-sectional area and diameter of muscle fiber were measured after HE staining. The mRNA expression of autophagy-realated factor Beclin-1, vacuolar protein sorting (Vps34) and microtubule-associated protein light chain 3 (LC3) were tested with reverse transcription real-time quantitative polymerase chain reaction.
Results There was no statistical difference in the ratio of gastrocnemius wet weight, the cross-sectional area and diameter of muscle fiber, and the mRNA expression of Beclin-1, Vps34 and LC3 among three groups on the 0th day (F < 1.321, P > 0.05). Compared with the sham group, the ratio of gastrocnemius wet weight, the cross-sectional area and diameter of muscle fiber decreased at different time points in the model group and Tuina group (P < 0.05), the ratio of gastrocnemius wet weight was higher, and the cross-sectional area and diameter of muscle fiber were bigger, both except on the 21st day, in Tuina group than in the model group (P < 0.05). Compared with the sham group, the mRNA expression of Beclin-1, Vps34 and LC3 increased at different points in the model group than in Tuina group (P < 0.05), and all the mRNA expression was higher, except on the 14th day, in Tuina group than in the model group (P < 0.05). The ratio of gastrocnemius wet weight, the cross-sectional area and diameter of muscle fiber showed a trend of progressive decrease with time in the model group and Tuina group (P < 0.05). The mRNA expression of Beclin-1 and Vps34 increased (P < 0.05), and the mRNA expression of LC3 increased in the model group 21 days after intervention (P < 0.05). The mRNA expression of Beclin-1, Vps34 and LC3 increased first and then decreased, except the mRNA expression on the 14th day in Tuina group (P < 0.05).
Conclusion Tuina may promote the activation of autophagy by up-regulating the expression of autophagy-realated factor Beclin-1, Vps34 and LC3, remove the damaged organelles and proteins, provide certain synthetic substrate and energy for muscle fiber regeneration, thereby reduce the loss of degree of denervated skeletal muscle atrophy.
AN Hui-yu
,
TANG Cheng-lin
,
HUANG Si-qin
,
ZHAO Dan-dan
,
LUO Ao
,
WU Meng-jia
,
TAN Cheng-fang
,
QIU Li
,
WAN Xiao-feng
,
MA Xiang
. Effect of Tuina on Autophagy-related Factor Beclin-1, Vacuolar Protein Sorting and Microtubule-associated Protein Light Chain 3 in Rats with Denervated Skeletal Muscle Atrophy[J]. Chinese Journal of Rehabilitation Theory and Practice, 2019
, 25(2)
: 184
-191
.
DOI: 10.3969/j.issn.1006-9771.2019.02.010
[1] Wineski L E, von Deutsch D A, Abukhalaf I K, et al. Muscle-specific effects of hindlimb suspension and clenbuterol in mature male rats [J]. Cells Tissues Organs, 2002, 171(2-3): 188-198.
[2] Stein T P, Wade C E. Protein turnover in atrophying muscle:from nutritional intervention to microarray expression analysis [J]. Curr Opin Clin Nutr Metab Care, 2003, 6(1): 95-102.
[3] O'Leary M F, Vainshtein A, Carter H N, et al. Denervation-induced mitochondrial dysfunction and autophagy in skeletal muscle of apoptosis-deficient animals [J]. Am J Physiol Cell Physiol, 2012, 303(4): C447-C454.
[4] Masiero E, Agatea L, Mammucari C, et al. Autophagy is required to maintain muscle mass [J]. Cell Metab, 2009, 10(6): 507-515.
[5] 马书杰,严隽陶,黄品贤. 推拿手法防治失神经骨骼肌萎缩的实验研究[J]. 江苏中医药, 2014, 46(11): 79-81.
[6] 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(5): 29-32.
[7] 吴梦佳,唐成林,赵丹丹,等. 电针对失神经大鼠骨骼肌自噬相关基因表达的影响[J]. 中国康复理论与实践, 2018, 24(3): 260-265.
[8] 高睿琦,唐成林,黄思琴,等. 电针对失坐骨神经大鼠腓肠肌细胞凋亡及相关蛋白的影响[J]. 针刺研究, 2017, 42(4): 302-307.
[9] 高睿琦,唐成林,曹净,等. 电针对失神经骨骼肌萎缩大鼠胰岛素样生长因子1、肌肉生长抑制素及肌卫星细胞增殖的影响[J]. 中国康复理论与实践, 2016, 22(11): 1259-1263.
[10] 赵建文,银欢,苏秋香. 大鼠肌卫星细胞移植失神经骨骼肌的形态学观测[J]. 中国比较医学杂志, 2009, 19(3): 50-52, 56, 90.
[11] 杨辉,唐成林,常青,等. 在大鼠骨骼肌急性损伤修复过程中按摩对神经型一氧化氮合成酶、肝细胞生长因子mRNA表达的影响[J].中国康复医学杂志, 2015, 30(3): 224-230.
[12] 金宏柱. 中国推拿[M]. 上海:上海中医药大学出版社, 2002: 16-33.
[13] 段斐,张华,吴越,等. 失神经骨骼肌萎缩的研究现状及进展[J]. 现代生物医学进展, 2017, 17(7): 1382-1386, 1334.
[14] 马肃霜,肖颖锋. 失神经肌萎缩的研究进展[J]. 华夏医学, 2013, 26(1): 198-202.
[15] 郭汝宝,翁军,李增图,等. 推拿手法对家兔失神经支配后肌球蛋白重链mRNA表达的影响[J]. 中华中医药学刊, 2015, 33(1): 46-48.
[16] 郭汝宝,张喜林,严隽陶. 推拿手法对家兔失神经支配骨骼肌复合动作电位及收缩功能的影响[J]. 中华中医药学刊, 2017, 35(1): 140-142.
[17] 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, 31(9): 1872-1874.
[18] 成军. 现代细胞自噬分子生物学[M]. 2版. 北京:科学出版社, 2016: 3.
[19] Pfeifer U.
[Morphological and functional aspects of cellular autophagy] [J].
[in German]. Acta Morphol Acad Sci Hung, 1972, 20(3): 247-267.
[20] Cao Y, Klionsky D J. Physiological functions of Atg6/Beclin 1:a unique autophagy-related protein [J]. Cell Res, 2007, 17(10): 839-849.
[21] Backer J M. The regulation and function of Class III PI3Ks:novel roles for Vps34 [J]. Biochem J, 2008, 410(1): 1-17.
[22] Dancourt J, Melia T J. Lipidation of the autophagy proteins LC3 and GABARAP is a membrane-curvature dependent process [J]. Autophagy, 2014, 10(8): 1470-1471.
[23] Wohlgemuth S E, Seo A Y, Marzetti E, et al. Skeletal muscle autophagy and apoptosis during aging: effects of calorie restriction and life-long exercise [J]. Exp Gerontol, 2010, 45(2): 138-148.
[24] 邱守涛,崔迪,付绍婷,等. 不同强度耐力运动对骨骼肌质量的影响:自噬与衰老[J]. 沈阳体育学院学报, 2015, 34(1): 78-84, 90.
[25] Lira V A, Okutsu M, Zhang M, et al. Autophagy is required for exercise training-induced skeletal muscle adaptation and improvement of physical performance [J]. FASEB J, 2013, 27(10): 4184-4193.
[26] MacKenzie M G, Hamilton D L, Murray J T, et al. mVps34 is activated following high-resistance contractions [J]. J Physiol, 2009, 587(1): 253-260.
[27] Feng Z, Bai L, Yan J, et al. Mitochondrial dynamic remodeling in strenuous exercise-induced muscle and mitochondrial dysfunction: regulatory effects of hydroxytyrosol [J]. Free Radic Biol Med, 2011, 50(10): 1437-1446.
