电针对创伤性脊髓损伤大鼠下肢骨骼肌萎缩相关蛋白表达的影响

doi:10.3969/j.issn.1006-9771.2019.10.004

《中国康复理论与实践》 ›› 2019, Vol. 25 ›› Issue (10): 1133-1139.doi: 10.3969/j.issn.1006-9771.2019.10.004

电针对创伤性脊髓损伤大鼠下肢骨骼肌萎缩相关蛋白表达的影响

范蕊^1,2, 吴宗辉^1,2, 陈晓琳^1,2, 邹佐强^1,2, 龙在云³, 姚兰^1,2, 李斌^1,2

1.西南大学医院，重庆市400715;
2.西南大学运动康复研究所，重庆市400715;
3.陆军军医大学大坪医院野战外科研究所第三研究室，创伤与烧伤国家重点实验室，重庆市 400042

出版日期:2019-10-25 发布日期:2019-10-30
通讯作者: 吴宗辉，E-mail： 185056720@qq.com E-mail:185056720@qq.com
作者简介:范蕊(1980-)，女，汉族，河南平顶山市人，硕士，主治医师，主要研究方向：物理因子治疗及运动损伤康复。
基金资助:
1.重庆市科委社会事业与民生保障科技创新项目(No. CSTC2016SHMZX130016)；2.中央高校基本科研基金项目(No. XDJK2017C090)；3.西南大学医院科研资金资助项目(No. xdyykyjj2019xm04)

Effects of Electroacupuncture on Skeletal Muscle Atrophy-associated Protein in Hind Limbs of Traumatic Spinal Cord Injury Rats

FAN Rui^1,2, WU Zong-hui^1,2, CHEN Xiao-lin^1,2, ZOU Zuo-qiang^1,2, LONG Zai-yun³, YAO Lan^1,2, LI Bin^1,2

1.Southwest University Hospital, Chongqing 400715, China;
2.Institute of Sports Rehabilitation, Southwest University, Chongqing 400715, China;
3.State Key Laboratory of Trauma, Burn and Combined Injury, Institute of Surgery Research, Daping Hospital, Army Medical University, Chongqing 400042, China

Published:2019-10-25 Online:2019-10-30
Contact: WU Zong-hui, E-mail: 185056720@qq.com E-mail:185056720@qq.com
Supported by:
Supported by Scientific and Technological Innovation Project of Social Career and People's Livelihood Guarantee of Chongqing Science and Technology Commission (No. CSTC2016SHMZX130016), The Fundamental Research Funds for the Central Universities (No. XDJK2017C090) and Southwest University Hospital Scientific Research Project (No. xdyykyjj2019xm04)

摘要/Abstract

摘要： 目的观察电针对创伤性脊髓损伤(TSCI)大鼠腓肠肌中肌肉生长抑制素(MSTN)、肌肉特异性环指蛋白1 (MuRF1/Trim63)、肌肉萎缩盒F蛋白32 (Atrogin-1/Fbxo32)、成肌分化抗原(Myod)和肌细胞生成素(Myog) mRNA表达的影响，探讨电针延缓TSCI大鼠下肢萎缩的可能机制。方法将雌性Sprague-Dawley大鼠45只随机分为假手术组(n = 12)和造模组(n = 33)。采用 Allen法制备大鼠TSCI模型。将造模组存活大鼠(n = 24)随机分为模型组(n = 12)和电针组(n = 12)。术后1 d，电针组电针大椎、命门和双侧足三里穴，共28 d。术前，术后3 d、7 d、14 d、21 d、28 d比较各组BBB评分；术前和术后28 d，比较各组大鼠体质量；术后28 d，比较各组腓肠肌湿重比，HE染色比较腓肠肌纤维截面积和直径，实时定量聚合酶链反应(PCR)比较各组腓肠肌中MSTN、Trim63、Fbxo32、Myod和Myog mRNA的相对表达量。结果术后3 d、7 d、14 d、21 d和28 d，模型组BBB评分明显低于假手术组(P < 0.01)，术后7 d、14 d、21 d和28 d，电针组BBB评分明显高于模型组(P < 0.01)。术后28 d，与假手术组比较，模型组体质量、左右侧肌湿重比、肌纤维横截面积和直径均减小(P < 0.01)，MSTN、Trim63、Fbxo32、Myod和Myog mRNA的相对表达量升高(P < 0.05)；与模型组比较，电针组左右侧肌湿重比、肌纤维横截面积和直径均增加(P < 0.05)，MSTN、Trim63和Fbxo32 mRNA相对表达量降低(P < 0.05)，Myod和Myog mRNA相对表达量升高(P < 0.05)。结论电针干预大鼠大椎、命门和双侧足三里穴可能通过下调MSTN、Trim63、Fbxo32 mRNA的表达，并上调Myod和Myog mRNA的表达，促进肌卫星细胞的成肌分化，延缓肌蛋白降解，从而减轻TSCI大鼠下肢腓肠肌萎缩。

关键词: 创伤性脊髓损伤, 肌萎缩, 电针, 肌肉生长抑制素, 肌卫星细胞, 大鼠

Abstract: Objective To explore the effects and mechanism of electroacupuncture (EA) on expression of myostatin (MSTN), muscle-specific ring finger protein 1 (MuRF1/Trim63), F-box only protein 32 (Atrogin-1/ Fbxo32), myogenic differentiation antigen (Myod) and myogenin (Myog) in traumatic spinal cord injury (TSCI) rats. Methods A total of 45 adult female Sprague-Dawley rats were randomly divided into sham operation group (n = 12) and operation group (n = 33). The TSCI model was established with the modified Allen's method. After modeling, there were 24 survival rats and they were randomly divided into model group (n = 12) and EA group (n = 12). EA group was electroacupunctured at Dazhui (DU 14), Mingmen (DU 4) and bilateral Zusanli (ST 36) for 10 minutes, once a day, six times a week for 28 days. Basso-Beattie-Bresnahan (BBB) score was tested before modeling, and three days, seven days, 14 days, 21 days and 28 days after modeling. The rats were measured their body mass before and 28 days after modeling. The ratio of gastrocnemius wet mass was calculated; the cross-sectional area (CSA) and fiber diameter were measured by HE staining; the expression of MSTN, Trim63, Fbxo32, Myod and Myog mRNA were tested with real-time quantitative polymerase chain reaction (qPCR). Results Three days, seven days, 14 days, 21 days, and 28 days after modeling, the score of BBB was lower in the model group than in the sham operation group (P < 0.01); seven days, 14 days, 21 days, and 28 days after modeling, the score of BBB was higher in EA group than in the model group (P < 0.01). Compared with the sham operation group, the mass of rats, the gastrocnemius wet mass, the CSA and the diameter of the muscle fiber were smaller in the model group (P < 0.05), while the expression of MSTN, Trim63, Fbxo32, Myod and Myog mRNA were higher (P < 0.05). Compared with the model group, the mass of rats, the gastrocnemius wet mass, the CSA, the expression of Myod and Myog mRNA were higher (P < 0.05) in EA group, while the expression of MSTN, Trim63 and Fbxo32 mRNA were lower (P < 0.05). Conclusion EA might delay the gastrocnemius atrophy in TSCI rats by down-regulating the expression of MSTN, Trim63, Fbxo32 mRNA and up-regulating the expression of Myod and Myog mRNA via controlling the differentiation of the muscle satellite cells and the degradation of protein in skeletal muscle cells.

Key words: traumatic spinal cord injury, muscle atrophy, electroacupuncture, myostatin, muscle satellite cells, rats

中图分类号:

R651.2

范蕊, 吴宗辉, 陈晓琳, 邹佐强, 龙在云, 姚兰, 李斌. 电针对创伤性脊髓损伤大鼠下肢骨骼肌萎缩相关蛋白表达的影响[J]. 《中国康复理论与实践》, 2019, 25(10): 1133-1139.

FAN Rui, WU Zong-hui, CHEN Xiao-lin, ZOU Zuo-qiang, LONG Zai-yun, YAO Lan, LI Bin. Effects of Electroacupuncture on Skeletal Muscle Atrophy-associated Protein in Hind Limbs of Traumatic Spinal Cord Injury Rats[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2019, 25(10): 1133-1139.

参考文献

1 KumarR, LimJ, MekaryR A, et al. Traumatic spinal injury: global epidemiology and worldwide volume [J]. World Neurosurg, 2018, 113: e345-e363.
2 陈星月,陈栋,陈春慧,等. 中国创伤性脊髓损伤流行病学和疾病经济负担的系统评价[J]. 中国循证医学杂志, 2018, 18(2): 143-150.
3 于铁强,冯世庆. 216例外伤性脊髓损伤住院患者流行病学分析[J]. 中国骨与关节损伤杂志, 2010, 25(7): 583-585.
4 SavikjM, RubyM A, KostovskiE, et al. Retained differentiation capacity of human skeletal muscle satellite cells from spinal cord-injured individuals [J]. Physiol Rep, 2018, 6(12): e13739.
5 FanzaniA, ConraadsV M, PennaF, et al. Molecular and cellular mechanisms of skeletal muscle atrophy: an update [J]. J Cachexia Sarcopenia Muscle, 2012, 3(3): 163-179.
6 O'BrienL C, GorgeyA S. Skeletal muscle mitochondrial health and spinal cord injury [J]. World J Orthop, 2016, 7(10): 628-637.
7 Yarar-FisherC, BickelC S, KellyN A, et al. Heightened TWEAK-NF-κB signaling and in?ammation -associated ?brosis in paralyzed muscles of men with chronic spinal cord injury [J]. Am J Physiol Endocrinol Metab, 2016, 10(9): E754-E761.
8 朱斌,王策,徐青青. 药物穴位注射联合夹脊督脉电针对脊髓损伤患者运动功能障碍改善的疗效观察[J]. 中国中医急症, 2017, 26(11): 2054-2056.
9 程瑞动,朱根应,叶祥明,等. 不同时期针刺对不完全性脊髓损伤患者盆底肌及排尿功能的影响[J]. 中国康复理论与实践, 2016, 22(12): 1438-1441.
10 闵友江,程立红,高洁. 三通针法治疗脊髓损伤恢复期截瘫患者临床观察[J]. 上海针灸杂志, 2013, 32(12): 1010-1013.
11 李波霖,周宾宾,金昕,等. 督脉对比夹脊电针治疗对脊髓损伤大鼠下肢功能恢复作用的网状Meta分析[J]. 中国中医基础医学杂志, 2016, 22(12): 1676-1680, 1716.
12 马睿杰,孙连珠,张柳娟,等. 电针结合减重步行训练对急性脊髓损伤大鼠巢蛋白和神经生长因子表达的影响[J]. 中国康复医学杂志, 2015, 30(6): 532-537.
13 曾欢欢,黄英如,李子健,等. 大黄素对大鼠急性脊髓损伤后继发脊髓水肿的影响[J]. 中国康复理论与实践, 2018, 24(4): 378-384.
14 赵丹丹,唐成林,黄思琴,等. 电针干预对大鼠腓肠肌适应性肥大及肌卫星细胞增殖分化的影响[J]. 针刺研究, 2017, 42(6): 489-495.
15 吕威,莫雨平,李冰,等. 督脉电针对不同时间段脊髓损伤大鼠运动功能及p75神经营养素受体表达的影响[J]. 中国康复理论与实践, 2016, 22(8): 876-883.
16 PeiJ P, FanL H, NanK, et al. HSYA alleviates secondary neuronal death through attenuating oxidative stress, inflammatory response, and neural apoptosis in SD rat spinal cord compression injury [J]. J Neuroinflammation, 2017, 14(1): 97-110.
17 KellerA V, WainwrightG, Shum-SiuA, et al. Disruption of locomotion in response to hindlimb muscle stretch at acute and chronic time points after a spinal cord injury in rats [J]. J Neurotrauma, 2017, 34(3): 661-670.
18 ZhangY T, JinH, WangJ H, et al. Tail nerve electrical stimulation and electro-acupuncture can protect spinal motor neurons and alleviate muscle atrophy after spinal cord transection in rats [J]. Neural Plast, 2017, 2017: 7351238.
19 ZhangC, RongW, ZhangG H, et al. Early electrical field stimulation prevents the loss of spinal cord anterior horn motoneurons and muscle atrophy following spinal cord injury [J]. Neural Regen Res, 2018, 13(5): 869-876.
20 王玲洁,王子礼,杜俊龙,等. 中医对脊髓损伤的认识及治疗探析[J]. 湖北民族学院学报(医学版), 2016, 33(4): 62-64.
21 谭明生. “病证结合,从督论治”颈脊髓损伤的思考[J]. 中国中医骨伤科杂志, 2018, 26(2): 1-2.
22 蒋松鹤,林海燕,何蓉,等. 督脉、夹脊电针对脊髓损伤大鼠功能康复的影响[J]. 中华针灸电子杂志, 2015, 4(1): 7-12.
23 HoogaarsW M H, JaspersR T. Past, present, and future perspective of targeting Myostatin and related signaling pathways to counteract muscle atrophy [J].Adv Exp Med Biol, 2018, 1088: 153-206.
24 BoonH, Sj?grenR J, MassartJ, et al. MicroRNA-208b progressively declines after spinal cord injury in humans and is inversely related to myostatin expression [J]. Physiol Rep, 2015, 3(11). pii: e12622.
25 TandoT, HirayamaA, FurukawaM, et al. Smad2/3 proteins are required for immobilization-induced skeletal muscle atrophy [J]. J Biol Chem, 2016, 291(23): 12184-12194.
26 FolettaV C, WhiteL J, LarsenA E, et al. The role and regulation of MAFbx/atrogin-1 and MuRF1 in skeletal muscle atrophy [J]. Pflugers Arch, 2011, 461(3): 325-335.
27 MendiasC L, LynchE B, GumucioJ P, et al. Changes in skeletal muscle and tendon structure and function following genetic inactivation of myostatin in rats [J]. J Physiol, 2015, 593(8): 2037-2052.
28 De GasperiR, GrahamZ A, HarlowL M, et al. The signature of MicroRNA dysregulation in muscle paralyzed by spinal cord injury includes downregulation of MicroRNAs that target Myostatin signaling [J]. PLoS One, 2016, 1(12): e0166189.
29 GuitartM, LloretaJ, Ma?as-GarciaL, et al. Muscle regeneration potential and satellite cell activation profile during recovery following hindlimb immobilization in mice [J]. J Cell Physiol, 2018, 233(5): 4360-4372.
30 GanassiM, BadodiS, Ortuste QuirogaH P, et al. Myogenin promotes myocyte fusion to balance fibre number and size [J]. Nat Commun, 2018, 9(1): 4232-4248.
31 李利军,宋洁富,魏杰,等. 慢性压迫性脊髓损伤后骨骼肌形态学及相关蛋白Myogenin和myoD表达的实验研究[J]. 中华临床医师杂志(电子版), 2010, 4(9): 1691-1694.

[1]	陈怡婷, 王倩, 崔慎红, 李映彩, 张思鈺, 魏衍旭, 任慧, 冷军, 陈斌. 双侧序贯重复经颅磁刺激干预脑卒中患者上肢运动功能的效果[J]. 《中国康复理论与实践》, 2023, 29(8): 926-932.
[2]	李芳, 霍速, 杜巨豹, 刘秀贞, 李小爽, 宋为群. 经颅直流电刺激联合任务导向性康复训练对脊髓损伤大鼠前肢运动障碍的效果[J]. 《中国康复理论与实践》, 2023, 29(7): 777-781.
[3]	罗兰, 李璐, 金沐. 氙气后处理对脊髓缺血再灌注损伤的效果：Akt信号通路和自噬机制[J]. 《中国康复理论与实践》, 2023, 29(2): 174-181.
[4]	黄志霖, 徐发邵, 施静, 黄淦, 刘梅芳, 张霞辉. 线栓法建立卒中后吞咽障碍的大鼠模型[J]. 《中国康复理论与实践》, 2023, 29(10): 1147-1153.
[5]	秦彦强, 董浩, 孙迎春, 程先宽, 姚海江. 不同针刺方案对卒中后抑郁大鼠神经递质及相关炎性因子的影响[J]. 《中国康复理论与实践》, 2023, 29(1): 30-37.
[6]	张立,吴珊红,赵乐乐,王艳. Netrin-1、Slit2在夹脊电针结合神经松动术调节兔坐骨神经损伤后Rho GTPases失衡的作用[J]. 《中国康复理论与实践》, 2022, 28(8): 914-926.
[7]	缪培,张通,米海霞,张伟东. 不同线栓法复制局灶性脑缺血模型大鼠恢复期学习记忆能力的差异及其机制[J]. 《中国康复理论与实践》, 2022, 28(7): 789-796.
[8]	周小珏,冯婧,庞日朝,刘捷,张安仁. 隔日限食减轻脊髓损伤大鼠炎症反应的芳香烃受体/细胞因子信号传导抑制因子2/核转录因子-κB信号通路机制[J]. 《中国康复理论与实践》, 2022, 28(5): 544-551.
[9]	宋绍霏,侯园园,王云雷,张通. 异常光周期所致昼夜节律紊乱对大鼠腓肠肌时钟基因和葡萄糖摄取相关基因表达节律的影响[J]. 《中国康复理论与实践》, 2022, 28(5): 552-558.
[10]	李童,方志鹏,邵玉萍,王平. 有氧运动对睡眠剥夺大鼠学习记忆及海马神经元突触可塑性的效果[J]. 《中国康复理论与实践》, 2022, 28(11): 1270-1277.
[11]	丁妍怡,张胜行,刘雨露,余燕,杨敏光,梁胜祥,柳维林,陶静. 电针对血管性认知障碍大鼠脑功能局部一致性的效果[J]. 《中国康复理论与实践》, 2022, 28(1): 55-61.
[12]	王静怡,尹杰,刘建成,庞日朝,王文春. 蜘蛛香环烯醚萜类成分对急性脊髓损伤大鼠神经细胞焦亡的影响[J]. 《中国康复理论与实践》, 2021, 27(6): 653-660.
[13]	周文美,陶陶,吴霜,王廷龙,杨正奕,张莹. 丰富环境对脑缺血再灌注损伤大鼠神经功能和缺血半暗带区葡萄糖代谢的影响[J]. 《中国康复理论与实践》, 2021, 27(5): 522-529.
[14]	王琼芬,王风波,王科,钟永强,王娇娇. 电针风池对急性脑梗死大鼠脑星形胶质细胞和神经元的保护效果[J]. 《中国康复理论与实践》, 2021, 27(3): 302-309.
[15]	李向哲,丁洁,王庆华,董传明,王彤,吴勤峰. 减重平板训练对脊髓损伤大鼠神经病理性疼痛及脊髓后角谷氨酸脱羧酶-65/67表达的影响[J]. 《中国康复理论与实践》, 2021, 27(2): 131-136.

电针对创伤性脊髓损伤大鼠下肢骨骼肌萎缩相关蛋白表达的影响

Effects of Electroacupuncture on Skeletal Muscle Atrophy-associated Protein in Hind Limbs of Traumatic Spinal Cord Injury Rats

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

Metrics

本文评价

推荐阅读 0