推拿对失神经肌萎缩大鼠肌特异性microRNA和肌卫星细胞增殖分化相关因子的影响

doi:10.3969/j.issn.1006-9771.2020.11.009

《中国康复理论与实践》 ›› 2020, Vol. 26 ›› Issue (11): 1297-1304.doi: 10.3969/j.issn.1006-9771.2020.11.009

推拿对失神经肌萎缩大鼠肌特异性microRNA和肌卫星细胞增殖分化相关因子的影响

马翔,唐成林(),赵丹丹,安荟羽,万小凤,谯童茜

重庆医科大学中医药学院,重庆市 400016

收稿日期:2019-11-13 修回日期:2020-02-14 出版日期:2020-11-25 发布日期:2020-11-24
通讯作者: 唐成林 E-mail:cytcl996@163.com
作者简介:马翔(1994-),男,汉族,新疆霍城县人,硕士研究生,主要研究方向：针灸推拿机制研究。|唐成林(1966-),男,硕士,教授,主要研究方向：针灸推拿机制研究。
基金资助:
1.国家自然科学基金项目(81273870);2.重庆市卫生和计划生育委员会、重庆市科学技术委员会联合资助中医药重点科研项目(ZY201801007)

Effects of Tuina on Muscle-specific microRNA and Factors Related with Proliferation and Differentiation of Muscle Satellite Cells in Denervation-induced Atrophy in Rats

MA Xiang,TANG Cheng-lin(),ZHAO Dan-dan,AN Hui-yu,WAN Xiao-feng,QIAO Tong-qian

College of Chinese Medicine of Chongqing Medical University, Chongqing 400016, China

Received:2019-11-13 Revised:2020-02-14 Published:2020-11-25 Online:2020-11-24
Contact: TANG Cheng-lin E-mail:cytcl996@163.com
Supported by:
National Natural Science Foundation of China(81273870);Chongqing Health and Family Planning Committee and Chongqing Science and Technology Committee Traditional Chinese Medicine Fund(ZY201801007)

摘要/Abstract

摘要：

目的探讨推拿对失神经骨骼肌萎缩的延缓效应及其可能机制。方法 42只雄性Sprague-Dawley大鼠随机分为假手术组(n = 6)、模型组(n = 18)和推拿组(n = 18)。假手术组游离右侧胫神经,模型组和推拿组游离并切除右侧胫神经约1 cm。术后第2天起,推拿组在损伤局部行推拿干预,假手术组和模型组仅固定不干预。模型组和推拿组分别于术后14 d、21 d、28 d各处死6只大鼠,假手术组术后28 d处死,取术侧腓肠肌测定湿重比,HE染色观察腓肠肌细胞直径和面积,逆转录实时定量聚合酶链反应检测各时间点配对盒转录因子(Pax7)、成肌分化抗原(MyoD)和肌细胞生成素(MyoG)表达,以及21 d时microRNA-1、microRNA-133a、microRNA-206的表达。结果与假手术组相比,模型组和推拿组各时间点腓肠肌湿重比、肌细胞直径、肌细胞面积(除14 d推拿组)均下降(P < 0.05);与模型组相比,推拿组各时间点腓肠肌湿重比、肌细胞直径、肌细胞面积均升高( P < 0.05)。与假手术组相比,模型组14 d时Pax7表达升高( P < 0.05)、28 d时表达降低( P < 0.05),推拿组各时间点(除28 d) Pax7表达明显升高( P < 0.05);与模型组相比,推拿组各时间点Pax7表达升高( P < 0.05)。与假手术组相比,模型组和推拿组各时间点MyoD、MyoG表达均升高( P < 0.05);与模型组相比,推拿组各时间点MyoD、MyoG (除14 d)表达均升高( P < 0.05)。21 d时,与假手术组相比,模型组和推拿组microRNA-1和microRNA-133a表达明显下降( P < 0.05),microRNA-206表达明显升高( P < 0.05);与模型组相比,推拿组microRNA-1、microRNA-133a和microRNA-206表达升高( P < 0.05)。结论推拿可能通过上调肌特异性microRNA的表达,促进Pax7/MyoD/MyoG通路转录,从而促进肌卫星细胞增殖分化,延缓失神经肌萎缩。

关键词: 失神经肌萎缩, 推拿, 肌卫星细胞, 增殖, 分化, microRNA, 大鼠

Abstract:

Objective To investigate the effects and mechanism of Tuina on denervation-induced atrophy.Methods A total of 42 Sprague-Dawley rats were randomly divided into sham group (n = 6), model group (n = 18) and Tuina group (n = 18). The model group and Tuina group freed and excised right tibia nerve about one centimeter, while the sham group freed the right tibia nerve only. From the second day after operation, Tuina group accepted Tuina on the injured area, while the sham group and the model group were only fixed without any intervention. Six rats were sacrificed on the 14th, 21st and 28th day after operation in the model and Tuina groups, and the sham group was sacrificed on the 28th day after operation. The gastrocnemius muscles were measured wet weight ratio. The diameter and area of muscle cells were measured under HE staining. The expression of Pax7, MyoD, MyoG, microRNA-1, microRNA-133a and microRNA-206 in the gastrocnemius muscles were detected with reverse transcription real-time quantitative polymerase chain reaction.Results Compared with the sham group, the wet weight ratio, the area of muscle cells (except the 14-day-Tuina group) and the diameter of muscle cells decreased at each time point in the model group and Tuina group (P < 0.05); compared with the model group, the wet weight ratio, muscle cell diameter and muscle cell area increased at each time point in Tuina group ( P < 0.05). Compared with the sham group, the expression of Pax7 increased in the 14-day-model group ( P < 0.05) and decreased in the 28-day-model group ( P < 0.05), and it increased at each time point (except 28-day) in Tuina group ( P < 0.05); compared with the model group, the expression of Pax7 increased at each time point in Tuina group ( P < 0.05). Compared with the sham group, the expression of MyoD and MyoG increased at each time point in the model group and Tuina group ( P < 0.05); compared with the model group, the expression of MyoD and MyoG increased at each time point (except 14-day) in Tuina group ( P < 0.05). Compared with the sham group, the expression of microRNA-1 and microRNA-133a decreased, and microRNA-206 increased in the model group and Tuina group at 21-day ( P < 0.05); compared with the model group, the expression of microRNA-1, microRNA-133a and microRNA-206 increased in Tuina group ( P < 0.05). Conclusion Tuina may activate the Pax7/MyoD/MyoG pathway by increasing the expression of muscle-specific microRNA, to promote the proliferation and differentiation of muscle satellite cells, and delay denervation-induced atrophy.

Key words: denervation-induced atrophy, Tuina, muscle satellite cells, proliferation, differentiation, microRNA, rats

中图分类号:

R685

马翔,唐成林,赵丹丹,安荟羽,万小凤,谯童茜. 推拿对失神经肌萎缩大鼠肌特异性microRNA和肌卫星细胞增殖分化相关因子的影响[J]. 《中国康复理论与实践》, 2020, 26(11): 1297-1304.

MA Xiang,TANG Cheng-lin,ZHAO Dan-dan,AN Hui-yu,WAN Xiao-feng,QIAO Tong-qian. Effects of Tuina on Muscle-specific microRNA and Factors Related with Proliferation and Differentiation of Muscle Satellite Cells in Denervation-induced Atrophy in Rats[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2020, 26(11): 1297-1304.

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组别	n	14 d	21 d	28 d
假手术组	6	1.00±0.02^*	1.00±0.02^*	1.00±0.02
模型组	6	0.58±0.07^a	0.38±0.03^a	0.25±0.02^a
推拿组	6	0.70±0.04^a,b	0.46±0.04^a,b	0.37±0.06^a,b
F值		115.220	865.980	677.010
P值		< 0.001	< 0.001	< 0.001

组别	n	14 d	21 d	28 d
假手术组	6	39.92±0.69^*	39.92±0.69^*	39.92±0.69
模型组	6	34.14±1.04^a	30.83±1.81^a	23.85±1.97^a
推拿组	6	38.11±1.33^a,b	32.86±0.96^a,b	26.93±0.71^a,b
F值		46.909	87.552	270.374
P值		< 0.001	< 0.001	< 0.001

组别	n	14 d	21 d	28 d
假手术组	6	1028.27±19.33^*	1028.27±19.33^*	1028.27±19.33
模型组	6	815.56±102.71^a	733.10±66.23^a	572.43±54.44^a
推拿组	6	967.50±79.14^b	868.17±15.67^a,b	653.08±37.16^a,b
F值		12.277	76.807	222.430
P值		0.001	< 0.001	< 0.001

组别	n	14 d	21 d	28 d
假手术组	6	1.01±0.12^*	1.01±0.12^*	1.01±0.12
模型组	6	1.83±0.15^a	1.06±0.44	0.50±0.21^a
推拿组	6	2.49±0.33^a,b	1.67±0.34^a,b	1.09±0.40^b
F值		67.533	7.651	8.293
P值		< 0.001	0.005	0.004

组别	n	14 d	21 d	28 d
假手术组	6	1.09±0.16^*	1.09±0.16^*	1.09±0.16
模型组	6	3.96±0.53^a	8.65±0.44^a	6.29±0.53^a
推拿组	6	4.65±0.34^a,b	11.63±0.50^a,b	7.49±0.29^a,b
F值		154.470	1132.584	533.664
P值		< 0.001	< 0.001	< 0.001

推拿对失神经肌萎缩大鼠肌特异性microRNA和肌卫星细胞增殖分化相关因子的影响

Effects of Tuina on Muscle-specific microRNA and Factors Related with Proliferation and Differentiation of Muscle Satellite Cells in Denervation-induced Atrophy in Rats

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组别	n	14 d	21 d	28 d
假手术组	6	1.01±0.15^*	1.01±0.15^*	1.01±0.15
模型组	6	21.65±1.49^a	33.73±1.48^a	13.36±0.77^a
推拿组	6	21.64±0.91^a	48.04±2.00^a,b	32.84±2.77^a,b
F值		833.071	1688.995	560.949
P值		< 0.001	< 0.001	< 0.001

组别	n	microRNA-133a	microRNA-206	microRNA-1
假手术组	6	1.03±0.18^*	1.05±0.31^*	1.01±0.13^*
模型组	6	0.21±0.06^a	3.11±0.51^a	0.59±0.11^a
推拿组	6	0.46±0.13^a,b	4.94±1.11^a,b	0.77±0.07^a,b
F值		57.763	42.764	23.095
P值		< 0.001	< 0.001	< 0.001

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