中国组织工程研究 ›› 2017, Vol. 21 ›› Issue (24): 3864-3869.doi: 10.3969/j.issn.2095-4344.2017.24.016

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

慢性膝关节炎疼痛模型的针灸治疗

徐丽丽1,黄 娟1,胥方元1,万永鲜2   

  1. 西南医科大学附属医院,1康复医学科,2骨与关节外科,四川省泸州市 646000
  • 修回日期:2017-06-07 出版日期:2017-08-28 发布日期:2017-08-30
  • 通讯作者: 万永鲜,博士,副教授,西南医科大学附属医院骨与关节外科,四川省泸州市 646000
  • 作者简介:徐丽丽,女,汉族,1981年生,山东省齐河县人,2004年成都体育学院运动医学毕业,康复主管技师,主要从事运动损伤康复研究。
  • 基金资助:

    四川省中医药管理局基金项目(2014-K-125);西南医科大学青年基金项目(2014QN-107);四川省卫计委普及应用项目(17PJ176)

Electroacupuncture for chronic pain in a model of knee arthritis

Xu Li-li1, Huang Juan1, Xu Fang-yuan1, Wan Yong-xian2   

  1. 1Department of Rehabilitation Medicine, 2Department of Bone and Joint Surgery, the Affiliated Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • Revised:2017-06-07 Online:2017-08-28 Published:2017-08-30
  • Contact: Wan Yong-xian, M.D., Associate professor, Department of Bone and Joint Surgery, the Affiliated Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • About author:Xu Li-li, Technician-in-charge, Department of Rehabilitation Medicine, the Affiliated Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • Supported by:

    the Project of Administration of Traditional Chinese Medicine of Sichuan, No. 2014-K-125; the Youth Foundation of Southwest Medical University, No. 2014QN-107; the Popularized Application Project of Health and Family Planning Commission of Sichuan Province, No. 17PJ176

摘要:

文章快速阅读:

文题释义:
白细胞介素(interleukin,IL):
最初是指由白细胞产生又在白细胞间发挥作用的细胞因子,虽然后来发现白细胞介素可由其他细胞产生, 也可作用于其他细胞,这一名称仍被沿用。在1979年第二届国际淋巴因子专题讨论会上,将来自单核-巨噬细胞、T淋巴细胞所分泌的某些非特异性发挥免疫调节和在炎症反应中起作用的因子称为白细胞介素(interleukin,IL)。目前发现了29 种白细胞介素,分别被命名为IL-1-IL-29。现在白细胞介素是指一类分子结构和生物学功能已基本明确,具有重要调节作用而统一命名的细胞因子,它和血细胞生长因子同属细胞因子。两者相互协调,相互作用,共同完成造血和免疫调节功能。白细胞介素在传递信息,激活与调节免疫细胞,介导T、B细胞活化、增殖与分化及在炎症反应中起重要作用。
白细胞介素17:是一种源于T细胞的细胞因子,有着广泛的生物学活性和参与机体免疫调节等作用,可通过复杂的作用机理调节细胞的生长、分化和增殖,诱导活性物质产生。

 

摘要
背景:
已经证实针灸对腰背痛、膝关节痛等急性疼痛等有较好疗效,但是对于慢性疼痛的效果存在争议。
目的:建立新西兰兔慢性关节炎疼痛模型,探讨针灸治疗慢性关节炎疼痛机制。  
方法:①32只新西兰兔使用4%木瓜蛋白酶0.3 mL注射双后肢膝关节腔内,制成兔膝骨性关节炎模型。随机分为4组,即生理盐水+电针组,生理盐水+假电针组,k-阿片受体拮抗剂(nor-Binaltorphimine,nor-BNI)+电针组,nor-BNI +假电针组,每组8只。nor-BNI用量 1 mg/kg,1次/d,连续3 d;电针干预时间为实验动物给予药物后2 h,1次/d,30 min/次,直至实验动物被处死当天结束;假电针组取穴、针刺方法及时间同电针组,但不通电刺激。使用BBB评分对各组进行下肢行为学评分。给药1,3,5,7日分别处死,分离脊髓组织后多聚甲醛固定,使用免疫荧光法检测脊髓组织白细胞介素17及白细胞介素17受体A、NR1的表达水平;②另取24只新西兰兔,12只兔建立膝骨性关节炎模型后随机分为模型+生理盐水组、模型+2 μg白细胞介素17抗血清组;剩余12只分为对照+生理盐水组、对照+2 μg白细胞介素17 抗血清组。以2 μg 白细胞介素17抗血清溶于10 μL生理盐水中进行椎管内给药,1次/d,连续3 d。使用BBB评分对新西兰兔下肢功能进行行为测试,每次行为测试后,Western-blot测定样品中 p-NR1,白细胞介素17R 的表达水平。
结果与结论:①nor-BNI +电针组BBB评分明显增高(P < 0.05);而脊髓组织白细胞介素17及白细胞介素17受体A、NR1的表达水平明显减低(P < 0.05);②使用nor-BNI拮抗阿片类受体后,nor-BNI+电针组脊髓组织NR1的表达水平与nor-BNI +假电针组相比无明显差异(P > 0.05);③使用白细胞介素17抗血清处理后,模型+2 μg白细胞介素17抗血清组BBB评分明显增高(P < 0.05);而该组脊髓组织白细胞介素17及NR1的表达水平明显减低(P < 0.05),但仍高于2个对照组(P < 0.05);④实验证实,慢性关节炎疼痛原因为白细胞介素17可以使脊髓组织NR1表达增加从而引起;电针刺激可以明显改善关节炎模型新西兰兔的下肢疼痛,这种作用是由于电针刺激减少脊髓组织白细胞介素17而非调节白细胞介素17受体A发挥作用。

 

关键词: 组织构建, 组织工程, 关节炎, 疼痛, 白细胞介素17, 阿片受体

Abstract:

BACKGROUND: Acupuncture has been found to be effective for alleviating low back pain and acute pain due to knee arthritis, but its effect on chronic pain is under discussion.

OBJECTIVE: To investigate the mechanism underlying electroacupuncture (EA) alleviating chronic pain in a New Zealand rabbit model of knee arthritis.
METHODS: (1) Thirty-two New Zealand rabbits were selected, and the knee osteoarthritis model was established by injecting 4% papain into the knee articular cavity of rabbit’s bilateral hind limbs. The model rabbits were randomly divided into four groups (n=8 per group): normal saline plus EA, normal saline plus sham EA, nor-Binaltorphimine (nor-BNI) plus EA, and nor-BNI plus sham EA groups. The dosage of nor-BNI was 1 mg/kg, once daily, for consecutive 3 days. 30-minute EA was given at 2 hours after administration, once daily, until the day the rabbits were killed. Sham EA indicated no given electric current. The behaviors of the lower limbs were evaluated by Basso, Beattie, Bresnahan scores. The rabbits were respectively killed at 1, 3, 5 and 7 days after administration, the spinal cord was separated, and then fixed with formaldehyde. The expression levels of interleukin-17, interleukin-17 receptor A and NR1 in the spinal cord tissues were detected by immunofluorescence. (2) The other 24 New Zealand rabbits were randomly divided into model and control groups (n=12 per group), and the knee osteoarthritis model was established in the former group. Afterwards, the two groups were randomized into two subgroups, followed by given the intrathecal administration of normal saline, or 2 μg interleukin-17 antibody serum dissolved in 10 μL normal saline, once daily, for consecutive 3 days. The behaviors of the lower limbs were evaluated by Basso, Beattie, Bresnahan scores, and the expression levels of p-NR1 and interleukin-17 receptor were detected by western blot assy.

RESULTS AND CONCLUSION: The Basso, Beattie, Bresnahan scores in the nor-BNI plus EA group were significantly increased, while the expression levels of interleukin-17, interleukin-17 receptor A and NR1 in the spinal cord tissues were significantly decreased (P < 0.05). The expression level of NRI did not differ significantly between nor-BNI plus EA and nor-BNI plus sham EA groups (P > 0.05). After administration of interleukin-17 antibody serum, the Basso, Beattie, Bresnahan scores in the model group was significantly increased, and the expression levels of interleukin-17 and NR1 in the spinal cord tissues were significantly decreased, but still significantly higher than those in the control subgroups (P < 0.05). These results suggest that chronic pain in knee arthritis is the result of an increase in the expression level of NRI induced by interleukin-17. EA can remarkably improve the pain in the model rabbits of knee arthritis by downregulating interleukin-17 in the spinal cord tissues, rather than interleukin-17 receptor.

 

Key words: Electroacupuncture, Arthritis, Pain, Tissue Engineering

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