主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

北京中医药大学学报 ›› 2019, Vol. 42 ›› Issue (7): 584-591.doi: 10.3969/j.issn.1006-2157.2019.07.010

• 中药药理 • 上一篇    下一篇

藏药宽筋藤对炎性疼痛大鼠血清中炎性细胞因子及脊髓中NMDA受体表达的影响*

马丹, 张硕峰#, 李颖慧, 程文豪, 宋敬怡, 程龙   

  1. 北京中医药大学中药学院 北京 102488
  • 收稿日期:2019-01-25 出版日期:2019-07-30 发布日期:2019-09-03
  • 通讯作者: 张硕峰,男,博士,教授,硕士生导师,主要研究方向:中药药性理论研究, E-mail: shuofengzhang@sina.com
  • 作者简介:马丹,女,在读硕士生
  • 基金资助:
    国家自然科学基金青年科学基金资助项目(No.81503287),中国医药教育协会孙思邈中医药科研专项项目(No.2016SKT-M006)

Effects of Tibetan medicine Tinospora sinensis on the expression of inflammatory cytokines and NMDA receptors in the spinal cord of rats with pain-sensitive hyperinjury*

Ma Dan, Zhang Shuofeng#, Li Yinghui, Cheng Wenhao, Song Jingyi, Cheng Long   

  1. School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China
  • Received:2019-01-25 Online:2019-07-30 Published:2019-09-03
  • Contact: Prof. Zhang Shuofeng, Ph.D., Master tutor. Beijing University of Chinese Medicine, School of Chinese Medicine, Beijing 102488. E-mail: shuofengzhang@sina.com

摘要: 目的 观察藏药宽筋藤水提物对炎性疼痛模型大鼠的镇痛作用,探究宽筋藤治疗炎性疼痛的作用机制。方法 大鼠按照随机数字法分为5组:正常组、模型组、醋酸泼尼松组、宽筋藤水提物高剂量组、宽筋藤水提物低剂量组,大鼠右后足注射完全弗氏佐剂造模24 h后,灌胃给药12 d,观察炎性疼痛模型大鼠的50%机械缩足痛反应值、热缩足反应时间和患足足跖肿胀率。末次给药1 h后灌注固定,取血清,ELISA方法测定白细胞介素-1α(IL-1α)、白细胞介素-6(IL-6)、白细胞介素-10(IL-10)、肿瘤坏死因子-α(TNF-α)的表达情况;取脊髓做免疫组化观察患侧背角N-甲基-D-天冬安酸(NMDA)受体表达情况。结果 模型组大鼠与正常组比较患足足跖部明显肿胀(P<0.01);宽筋藤水提物高剂量(2.4 g/kg)可明显降低大鼠足跖的肿胀程度,与各时间点模型组比较差异有统计学意义(P<0.01)。模型大鼠热缩足反应时间明显降低,与正常组比较差异有统计学意义(P<0.01);宽筋藤水提物不同剂量在各时间点均可明显延长大鼠热缩足反应时间,明显降低患肢处痛觉敏感程度,与模型组比较差异有统计学意义(P<0.01)。模型大鼠50%机械缩足反应值与对照组比较明显降低(P<0.01),宽筋藤给药组连续给药12 d期间内持续产生镇痛效果,与正常组比较差异有统计学意义(P<0.01)。与正常组比较,模型大鼠血清炎症细胞因子IL-1α、TNF-α、IL-6含量明显增高(P<0.01),IL-10含量明显降低(P<0.01);给药12 d后,宽筋藤水提物可明显降低模型大鼠血清IL-1α、TNF-α、IL-6含量并升高模型大鼠IL-10含量,与模型组比较差异有统计学意义(P<0.01)。与正常组比较,炎性疼痛模型大鼠损伤侧的脊髓背角中NMDA受体表达增加(P<0.01);与模型组比较,宽筋藤水提物低、高剂量可明显降低炎性疼痛模型大鼠患侧脊髓中NMDA受体表达(P<0.01)。结论 宽筋藤水提物可有效降低炎性疼痛大鼠对于温度和机械刺激的高敏感性,其抗炎镇痛机制与其降低外周炎性因子的产生及抑制NMDA受体表达有关。

关键词: 宽筋藤, 炎性疼痛, N-甲基-D-天冬氨酸受体, 炎性因子, 大鼠

Abstract: Objective To observe the effect of water extract of Tinospora sinensis on hypersensitivity to temperature and mechanical stimulation in rats with inflammatory pain and its anti-inflammatory and analgesic mechanism. The expression levels of IL-1α in serum, IL-6, IL-10, TNF-α and N-methyl-D-aspartic acid (NMDA) receptors in the spinal dorsal horn of the affected side were measured to explore the anti-inflammatory and analgesic mechanisms of Tinospora sinensis in the treatment of inflammatory pain in rats. Methods Rats were randomly divided into five groups by using randomized sequence table: normal group, model group, prednisone acetate group, high-dose, and mid-dose Tinospora sinensis water extract (TSWE) groups. After 24 hours of Freund’s adjuvant injection to establish the inflammatory model, the rats were intragastrically administered for 12 days. The reaction value of 50% mechanical foot contraction pain, reaction time of thermal foot contraction and swelling rate of affected foot and sole were then evaluated. After 1 day of the last administration, the rats were sacrificed with feet perfused and fixed. ELISA method was used to determine the expression of IL-1α, IL-6, IL-10 and TNF-α. The spinal cord was taken out for immunohistochemistry to observe the expression of NMDA receptor in the dorsal horn of the affected side. Results Compared with the control group, rats in the model group developed significant swelling in the ankle and feet (P<0.01). The 2.4 g/kg Tinospora sinensis water extract significantly reduced the degree of swelling at day 12 of administration (P<0.01, compared with the model group, P<0.05). The reaction time of heat-shrinking foot in model rats was significantly reduced and there was significant difference between the two groups (P<0.01, compared with the control group). Different doses of Tinospora sinensis could obviously increase the reaction time of rat’s heat-shrinking foot and prolong the pain-sensitive reaction of affected limbs at different time points, which had statistical significance (P<0.01, compared with the model group). The 50 % mechanical pain response value of the model rats was significantly lower than that of control group (P<0.01). The analgesic effect of Tinospora sinensis group lasted for 12 days, which was significantly different from that of the normal group (P<0.01). Compared with the normal group, serum inflammatory cytokines IL-1α, TNF-α and IL-6 in the model rats were significantly increased (P<0.01), and IL-10 was significantly decreased (P<0.01). After 12 days of administration, Tinospora sinensis water extract could significantly reduce the IL-1α, TNF-α and IL-6 and increase the IL-10 in the model rats (P<0.01, compared with the normal group). Compared with the normal group, the expression of NMDA receptor in the spinal dorsal horn of the injured side of the inflammatory pain model rats increased (P<0.01). Compared with the model group, 1.2, 2.4 g/kg Tinospora sinensis water extract could significantly reduce the expression of NMDA receptor in the spinal cord of rats with inflammatory pain model (P<0.01). Conclusion Tinospora sinensis water extract can effectively reduce the high sensitivity of temperature and mechanical stimulation in rats with inflammatory pain, and its anti-inflammatory and analgesic mechanism is related to reducing the production of peripheral inflammatory factors and inhibiting the expression of NMDA receptors.

Key words: Tinospora sinensis, inflammatory pain, NMDA, inflammatory factors, rats

中图分类号: 

  • R285.5