国际口腔医学杂志 ›› 2020, Vol. 47 ›› Issue (6): 635-643.doi: 10.7518/gjkq.2020102

• 论著 • 上一篇    下一篇

穴位中频脉冲电刺激对下颌前伸大鼠咬肌改建的生理与生化研究

陈青青(),刘珍巧,王豫蓉()   

  1. 重庆医科大学附属口腔医院正畸科 口腔疾病与生物医学重庆市重点实验室 重庆市高校市级口腔生物医学工程重点实验室 重庆 401147
  • 收稿日期:2019-12-05 修回日期:2020-06-10 出版日期:2020-11-01 发布日期:2020-11-06
  • 通讯作者: 王豫蓉
  • 作者简介:陈青青,硕士,Email: 2017110898@stu.cqmu.edu.cn
  • 基金资助:
    重庆市卫生健康委员会中医药科技项目(2019ZY023110);重庆高校创新团队建设计划(CXTDG201602006)

Physiological and biochemical study of acupoint mid-frequency pulse electrical stimulation on masseter muscle adapta-tions in rats with mandibular

Chen Qingqing(),Liu Zhenqiao,Wang Yurong()   

  1. Dept. of Orthodontics, Stomatological Hospital of Chongqing Medical University, Chongqing Key Laboratory of Oral Diseases and Biomedical Science, Chong-qing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Chongqing 401147, China
  • Received:2019-12-05 Revised:2020-06-10 Online:2020-11-01 Published:2020-11-06
  • Contact: Yurong Wang
  • Supported by:
    Science and Technology Project of Traditional Chinese Medicine of Chongqing Health Com-mittee(2019ZY023110);Chongqing University Innovation Team Construction Plan(CXTDG201602006)

摘要:

目的 研究穴位中频脉冲电刺激对下颌前伸大鼠咬肌肌电和脑源性神经营养因子(BDNF)表达的影响。方法 60只雄性Sprague-Dawley大鼠随机分为3个大组(实验组、条件对照组和空白对照组),每个大组根据实验观测天数又随机分为4个小组(3、7、14、21 d组),共12组(n=5)。实验组戴用前导下颌装置且接受穴位中频脉冲电刺激;条件对照组戴用前导下颌装置但不接受穴位中频脉冲电刺激;空白对照组不戴用前导下颌装置也不接受穴位中频脉冲电刺激。选择大鼠双侧颊车穴进行中频脉冲电刺激,参数设置为:正弦波形,0~150 Hz调制频率,2 000~8 000 Hz中频电频率。采用BL-420F生物信号采集与分析系统记录各组大鼠在不同实验时期(3、7、14、21 d)静息状态下的咬肌肌电,采用免疫组织化学染色法检测各组大鼠咬肌中BDNF蛋白的表达情况,采用实时荧光定量聚合酶链反应检测各组大鼠咬肌中BDNF mRNA的相对表达量。结果 实验组与条件对照组的咬肌肌电与BDNF蛋白和mRNA表达在观察期间均呈先上升后下降的趋势;3 d时,实验组咬肌肌电和BDNF蛋白表达水平均较条件对照组高(P<0.05);21 d时,实验组咬肌肌电和BDNF蛋白表达水平已基本恢复至正常水平,而条件对照组仍高于空白对照组(P<0.05)。结论 穴位中频脉冲电刺激可早期上调下颌前伸大鼠咬肌BDNF的表达,加快咬肌肌电恢复至正常水平的速度,促进大鼠咬肌适应性改建。

关键词: 电刺激, 下颌前伸, 咬肌, 肌电, 脑源性神经营养因子

Abstract:

Objective This study aimed to investigate the effects of acupoint mid-frequency pulse electrical stimulation on the surface electromyography and the expression of brain-derived neurotropic factor (BDNF) in the mandibular protrusion of Sprague-Dawley (SD) rats. Methods Sixty male SD rats were randomly divided into three large groups, namely, blank control, conditional control (with functional appliance and without acupoint mid-frequency pulse electrical stimulation therapy) and experimental groups (with functional appliance and acupoint mid-frequ- ency pulse electrical stimulation therapy). Each large group was randomly divided into four groups (3, 7, 14 and 21 days according to the number of experimental observation days), resulting in 12 groups (n=5). In this experiment, acupuncture points were selected on both sides of the rat, and the parame-ters of acupoint mid-frequency pulse electrical stimulation therapy were set as follows: sinusoidal waveform, 0-150 Hz modulation frequency, and 2 000-8 000 Hz intermediate frequency electrical frequency. The BL-420F biological signal acquisition and analysis system was used to record the electromyograms of rats in the resting state at different experimental periods (3, 7, 14, and 21 days). Immunohistochemistry and real-time quantitative polymerase chain reaction were carried out to detect the protein and mRNA expression levels of BDNF in the masseter muscle of rats. Results The electromyo-grams of masseter muscle, the protein and mRNA expression levels of BDNF first increased and then decreased in the experimental and condition control groups. Moreover, the electromyography of masseter muscle, the protein and mRNA expression levels of BDNF in the experimental group were higher than those in the condition control group on day 3 (P<0.05); the electromyography of masseter and the protein and mRNA expression levels of BDNF in the experimental group basically returned to the normal level on day 21, while those in the condition control group remained higher than those in the blank control group (P<0.05). Conclusion Acupoint mid-frequency pulse electrical stimulation therapy can improve the expression of BDNF in the masseter muscle of rats with functional mandibular protrusion in the early stage, accelerate the recovery of the masseter muscle to normal level, and may improve the treatment of functional appliance.

Key words: electric stimulation, mandibular protrusion, masseter muscle, electromyographic, brain-derived neuro-tropic factor

中图分类号: 

  • R78

图1

颊车穴定位"

表1

qRT-PCR引物序列"

引物名称 引物序列(5’—3’)
β-actin F:TCATGAAGTGTGACGTGGACATC
R:TGTTGCATTTGCGGGGACGATG
BDNF F:GAGCTGAGCGTGTGTGACAG
R:CGCCAGCCAATTCTCTTTTTGC

图2

各组大鼠下颌姿势位咬肌静息肌电变化"

表2

各组大鼠下颌姿势位咬肌静息积分肌电值"

戴入时间 空白对照组 条件对照组 实验组
戴入前 143.81±5.20 147.71±4.27 150.61±2.41
戴入当时 149.40±5.83 214.71±4.79a 241.13±6.85ab
戴入3 d 142.43±7.65 202.67±5.89a 228.40±7.58ab
戴入7 d 149.99±2.13 188.16±3.92a 204.66±6.89ab
戴入14 d 144.44±5.46 178.74±3.65a 166.43±5.03ab
戴入21 d 148.59±3.15 175.49±2.65a 155.11±7.44b

图3

各组大鼠下颌姿势位时咬肌的组织学变化 HE × 400 上:空白对照组;中:条件对照组;下:实验组。"

图4

各组大鼠咬肌BDNF蛋白的表达变化 SABC × 400 上:空白对照组;中:条件对照组;下:实验组。"

表3

大鼠咬肌BDNF蛋白的IOD测量值变化"

戴入时间/d 空白对照组 条件对照组 实验组
3 158.16±15.64 167.50±5.96 183.69±16.88ab
7 156.49±12.22 225.20±10.54a 213.43±9.49a
14 150.53±6.94 229.14±9.68a 196.45±13.11ab
21 149.33±5.64 200.93±20.25a 155.63±10.09b

表4

各组大鼠咬肌BDNF mRNA的相对表达量"

戴入时间/d 空白对照组 条件对照组 实验组
3 1.03±0.22 1.29±0.13 1.68±0.30ab
7 1.02±0.0.14 1.62±0.29a 1.43±0.20a
14 1.01±0.14 1.47±0.17a 1.36±0.34a
21 1.01±0.20 1.31±0.28a 1.17±0.33b
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