EFFECT OF PASSIVE MOVEMENT ON EXPRESSION OF miRNA-1 AND DIFFERENTIATION OF MYOBLASTS IN DENERVATION-INDUCED SKELETAL MUSCLE ATROPHY IN RATS_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

LIUZeyuan ¹ , ZHANGWenping ¹ , HUANGQiangkai ¹ , LIGang ² , CHENZhi ² , WEIJian ² ,  LIANGBingsheng ²

1. The Second Clinical Medical College of Shanxi Medical University, Taiyuan Shanxi, 030001, P. R. China;
2. Department of Orthopedics, the Second Hospital of Shanxi Medical University;

Corresponding author：

LIANGBingsheng, Email: liangbs707@aliyun.com

Keywords：

Denervation; Muscle atrophy; Passive movement; miRNA-1; Myocyte differentiation factor; Cell differentiation; Rat

DOI：

10.7507/1002-1892.20160124

Video：

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Objective To investigate the expression of miRNA-1 in denervated skeletal muscle at different periods, and to explore effects of passive movement on the expression of miRNA-1 and differentiation of myoblasts in denervation-induced skeletal muscle atrophy in rats. Methods Twenty-seven Sprague Dawley rats, weighing (200±10) g, were randomly divided into sham-operated group (group A, n=3), denervated group (group B, n=12), and passive movement group (group C, n=12). After the right sciatic nerve was exposed and dissociated, the sciatic nerve of 1 cm in length was removed in groups B and C; resection was not performed in group A. At 1 day after operation, passive flexion and extension movement was performed on the right hind limb in group C. At 6 hours in group A and at 3, 7, 14, and 28 days in groups B and C, 3 rats were sacrificed to measure the wet weight ratio of gastrocnemius muscle, to observe the diameter of the gastrocnemius muscle cell and evaluate the muscle atrophy by HE staining; RT-PCR was used to detect the mRNA expression of miRNA-1 and myocyte differentiation factor (MyoD), and immunohistochemistry to determine the protein expression of MyoD. Results Atrophy in various degrees was observed in denervated gastrocnemius muscle of groups B and C. The muscle fiber arranged in disorder and the diameter of the muscle cells decreased gradually with the time, without normal structure and morphology. The wet weight ratio and the cell diameter of the gastrocnemius in groups B and C were significantly less than those in group A (P<0.05); the wet weight ratio at 7, 14, 28 days and the cell diameter at 7, 14 days of group B were significantly greater than those of group A (P<0.05). The expressions of miRNA-1 and MyoD mRNA gradually increased with time in groups B and C, but were significantly less than those of group A at each time point (P<0.05). At 7, 14, and 28 days after operation, the expressions of miRNA-1 and MyoD mRNA in group C were significantly higher than those in group B (P<0.05). Immunohistochemical staining showed positive expression of MyoD in groups A, B, and C at each time point, but higher expression was observed in groups B and C than group A; the expression increased with time in groups B and C, and it was significantly higher in group C than group B. The correlation analysis results showed that the overall change trend of miRNA-1 and MyoD had no relation with the gastrocnemius wet weight ratio at 3 and 7 days (P>0.05), and had positive correlation at 14 and 28 days (P<0.05); positive correlation was found between the relative expression of MyoD and miRNA-1 mRNA (P<0.05). Conclusion Passive movement can prevent amyotrophy by increasing the expression of miRNA-1 and promoting the differentiation of myoblasts.

Citation： LIUZeyuan, ZHANGWenping, HUANGQiangkai, LIGang, CHENZhi, WEIJian, LIANGBingsheng. EFFECT OF PASSIVE MOVEMENT ON EXPRESSION OF miRNA-1 AND DIFFERENTIATION OF MYOBLASTS IN DENERVATION-INDUCED SKELETAL MUSCLE ATROPHY IN RATS. Chinese Journal of Reparative and Reconstructive Surgery, 2016, 30(5): 612-618. doi: 10.7507/1002-1892.20160124 Copy

1.	Goljanek-Whysall K, Sweetman D, Mtinsterberg AE. microRNAs in skeletal muscle differentiation and disease. Clin Sci (Lond), 2012, 123(11):611-625.
2.	Safdar A, Abadi A, Akhtar M, et al. miRNA in the regulation of skeletal muscle adaptation to acute endurance exercise in C57Bl/6J male mice. PLoS One, 2009, 4(5):e5610.
3.	Nielsen S, Scheele C, Yfanti C, et al. Muscle specifi cmicroRNAs are regulated by endurance exercise in human skeletal muscle. J Physiol, 2010, 588(Pt 20):4029-4037.
4.	王静, 宋新磊, 崔焕先, 等. 骨骼肌卫星细胞的特性及其增殖与分化的调控. 细胞生物学杂志, 2007, 29(4):497-502.
5.	Townley-Tilson WH, Callis TE, Wang D. MicroRNAs 1, 133, and 206:critical factors of skeletal and cardiac muscle development, function, and disease. Int J Biochem Cell Biol, 2010, 42(8):1252-1255.
6.	Koutsoulidou A, Mastroyiannopoulos NP, Furling D, et al. Expression of miR-1, miR-133a, miR-133b and miR-206 increases during development of human skeletal muscle. BMC Dev Biol, 2011, 11:34.
7.	冯小宁, 李文斌, 李永平, 等. MicroRNA-1/133a在大鼠失神经骨骼肌中的初步研究. 中华显微外科杂志, 2014, 37(1):56-62.
8.	弓贺炜, 李波, 李文斌, 等. miRNA-1和miRNA-133过表达对L6细胞增殖与分化的影响. 中华细胞与干细胞杂志, 2014, 4(2):89-96.
9.	Gayraud-Morel B, Chrétien F, Jory A, et al. Myf5 haploinsuffciency reveals distinct cell fate potentials for adult skeletal muscle stem cells. J Cell Sci, 2012, 125(Pt 7):1738-1749.
10.	Farina NH, Hausburg M, Betta ND, et al. A role for RNA posttranscriptional regulation in satellite cell activation. Skelet Muscle, 2012, 2(1):21.
11.	Antunes-Foschini RS, Miyashita D, Bicas HE, et al. Activated satellite cells in medial rectus muscles of patients with strabismus. Invest Ophthalmol Vis Sci, 2008, 49(1):215-220.
12.	Parker MH, von Maltzahn J, Bakkar N, et al. MyoD-dependent regulation of NF-κB activity couples cell-cycle withdrawal to myogenic differentiation. Skelet Muscle, 2012, 2(1):6.
13.	Ishido M, Kami K, Masuhara M. In vivo expression patterns of MyoD, p21, and Rb proteins in myonuclei and satellite cells of denervated rat skeletal muscle. Am J Physiol Cell Physiol, 2004, 287(2):484-493.
14.	L'honore A, Lamb NJ, Vandromme M, et al. MyoD distal regulatory region contains an SRF binding CArG element required for MyoD expression in skeletal myoblasts and during muscle regeneration. Mol Biol Cell, 2003, 14(5):2151-2162.
15.	Pietrangelo T, Di Filippo ES, Mancinelli R, et al. Low intensity exercise training improves skeletal muscle regeneration potential. Front Physiol, 2015, 6:399.
16.	谢永涛, 蓝岚. 骨骼肌卫星细胞生长因子与运动训练的关系. 中国组织工程研究与临床康复, 2011, 15(37):7009-7012.
17.	Sousa-Victor P, Muoz-Cánoves P, Perdiguero E. Regulation of skeletal muscle stem cells through epigenetic mechanisms. Toxicol Mech Methods, 2011, 21(4):334-342.
18.	Dey BK, Gagan J, Dutta A. miR-206 and-486 induce myoblast differentiation by downregulating Pax7. Mol Cell Biol, 2011, 31(1):203-214.
19.	Hirai H, Verma M, Watanabe S, et al. MyoD regulates apoptosis of myoblasts through microRNA-mediated down-regulation of Pax. J Cell Biol, 2010, 191(2):347-365.
20.	Zhang T, Birbrair A, Wang ZM, et al. Improved knee extensor strength with resistance training associateswith muscle specific miRNAs in older adults. Exp Gerontol, 2015, 62:7-13.

1. Goljanek-Whysall K, Sweetman D, Mtinsterberg AE. microRNAs in skeletal muscle differentiation and disease. Clin Sci (Lond), 2012, 123(11):611-625.
2. Safdar A, Abadi A, Akhtar M, et al. miRNA in the regulation of skeletal muscle adaptation to acute endurance exercise in C57Bl/6J male mice. PLoS One, 2009, 4(5):e5610.
3. Nielsen S, Scheele C, Yfanti C, et al. Muscle specifi cmicroRNAs are regulated by endurance exercise in human skeletal muscle. J Physiol, 2010, 588(Pt 20):4029-4037.
4. 王静, 宋新磊, 崔焕先, 等. 骨骼肌卫星细胞的特性及其增殖与分化的调控. 细胞生物学杂志, 2007, 29(4):497-502.
5. Townley-Tilson WH, Callis TE, Wang D. MicroRNAs 1, 133, and 206:critical factors of skeletal and cardiac muscle development, function, and disease. Int J Biochem Cell Biol, 2010, 42(8):1252-1255.
6. Koutsoulidou A, Mastroyiannopoulos NP, Furling D, et al. Expression of miR-1, miR-133a, miR-133b and miR-206 increases during development of human skeletal muscle. BMC Dev Biol, 2011, 11:34.
7. 冯小宁, 李文斌, 李永平, 等. MicroRNA-1/133a在大鼠失神经骨骼肌中的初步研究. 中华显微外科杂志, 2014, 37(1):56-62.
8. 弓贺炜, 李波, 李文斌, 等. miRNA-1和miRNA-133过表达对L6细胞增殖与分化的影响. 中华细胞与干细胞杂志, 2014, 4(2):89-96.
9. Gayraud-Morel B, Chrétien F, Jory A, et al. Myf5 haploinsuffciency reveals distinct cell fate potentials for adult skeletal muscle stem cells. J Cell Sci, 2012, 125(Pt 7):1738-1749.
10. Farina NH, Hausburg M, Betta ND, et al. A role for RNA posttranscriptional regulation in satellite cell activation. Skelet Muscle, 2012, 2(1):21.
11. Antunes-Foschini RS, Miyashita D, Bicas HE, et al. Activated satellite cells in medial rectus muscles of patients with strabismus. Invest Ophthalmol Vis Sci, 2008, 49(1):215-220.
12. Parker MH, von Maltzahn J, Bakkar N, et al. MyoD-dependent regulation of NF-κB activity couples cell-cycle withdrawal to myogenic differentiation. Skelet Muscle, 2012, 2(1):6.
13. Ishido M, Kami K, Masuhara M. In vivo expression patterns of MyoD, p21, and Rb proteins in myonuclei and satellite cells of denervated rat skeletal muscle. Am J Physiol Cell Physiol, 2004, 287(2):484-493.
14. L'honore A, Lamb NJ, Vandromme M, et al. MyoD distal regulatory region contains an SRF binding CArG element required for MyoD expression in skeletal myoblasts and during muscle regeneration. Mol Biol Cell, 2003, 14(5):2151-2162.
15. Pietrangelo T, Di Filippo ES, Mancinelli R, et al. Low intensity exercise training improves skeletal muscle regeneration potential. Front Physiol, 2015, 6:399.
16. 谢永涛, 蓝岚. 骨骼肌卫星细胞生长因子与运动训练的关系. 中国组织工程研究与临床康复, 2011, 15(37):7009-7012.
17. Sousa-Victor P, Muoz-Cánoves P, Perdiguero E. Regulation of skeletal muscle stem cells through epigenetic mechanisms. Toxicol Mech Methods, 2011, 21(4):334-342.
18. Dey BK, Gagan J, Dutta A. miR-206 and-486 induce myoblast differentiation by downregulating Pax7. Mol Cell Biol, 2011, 31(1):203-214.
19. Hirai H, Verma M, Watanabe S, et al. MyoD regulates apoptosis of myoblasts through microRNA-mediated down-regulation of Pax. J Cell Biol, 2010, 191(2):347-365.
20. Zhang T, Birbrair A, Wang ZM, et al. Improved knee extensor strength with resistance training associateswith muscle specific miRNAs in older adults. Exp Gerontol, 2015, 62:7-13.

Chinese Journal of Reparative and Reconstructive Surgery

EFFECT OF PASSIVE MOVEMENT ON EXPRESSION OF miRNA-1 AND DIFFERENTIATION OF MYOBLASTS IN DENERVATION-INDUCED SKELETAL MUSCLE ATROPHY IN RATS

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