腺苷A3受体在中枢神经系统痛觉传导通路及浅筋膜的表达分布
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摘要
目的:明确腺苷A3受体在针刺镇痛中的作用,探讨腺苷A3受体在中枢神经系统痛觉传导通路及浅筋膜的表达分布。方法:免疫组织化学法及Western Blot法分析腺苷A3受体在大鼠L4-S1脊髓神经节段、脑干、丘脑、大脑皮层及后三里、腹股沟中点处、腹膜浅筋膜的表达分布;鼠尾光照测痛仪观察针刺大鼠后三里对热辐射痛的镇痛作用,腹腔注射IB-MECA及Reversine对针刺镇痛的影响。结果:免疫组织化学法及Western Blot法分析显示,腺苷A3受体在痛觉传导通路中的脊髓、脑干、丘脑、大脑皮层均有分布,且在脊髓表达最高,依次是丘脑,大脑皮层,脑干。腺苷A3受体在后三里、腹股沟中点处、腹膜浅筋膜亦有分布,Western Blot法分析腹膜表达高于腹股沟和后三里,但免疫组织化学法分析各组光密度这一趋势并不明显。针刺后三里穴位进针后5、10、15、20、25、30 min甩尾潜伏期明显延长,具有时间依赖性,且在进针15～20 min镇痛效果最佳。腺苷A3受体激动剂IB-MECA可加强针刺效应,而腺苷A3受体拮抗剂Reversine可减弱该效应。结论:腺苷A3受体在中枢神经系统痛觉传导通路及浅筋膜均有丰富表达,参与针刺镇痛生理过程。
Objective:To investigate the role of adenosine A3 receptor in acupuncture analgesia and explore the expression and distribution of adenosine A3 receptor in the central nervous system pain conduction pathway and the superficial fascia.Methods:Immunohistochemistry and Western blot analysis were used for adenosine A3 receptor in rat L4-S1 spinal segment, brainstem, thalamus, cerebral cortex and after three years, at the midpoint of inguinal and peritoneal fascia expression. Rat tail light dolorimeter was used to analyze the analgesic effect of acupuncture at "Housanli" on thermal radiation pain. Effects of intraperitoneal injection of IB-MECA and Reversine on acupuncture analgesia were tested.Results:Immunohistochemistry and Western blot analysis showed that adenosine A3 receptors were distributed in the spinal cord, brainstem, thalamus and cerebral cortex in the pain transmission pathway, and the highest expression was in the spinal cord, followed by thalamus, cerebral cortex and brainstem. The adenosine A3 receptor was also distributed in "Housanli", the inguinal midpoint and the superficial peritoneum fascia. Western blot analysis showed that the peritoneal expression was higher than that in the groin and "Housanli", but the trend of light density in each group was not obvious by immunohistochemical analysis. After acupuncture at "Housanli" for 5 min, 10 min, 15 min, 20 min, 25 min, 30 min,tail flick latency was significantly prolonged, with time dependence, and needling for 15 min to 20 min had the best analgesic effect. Adenosine A3 receptor agonist IB-MECA enhanced the acupuncture effect while adenosine A3 receptor antagonist reversine can weaken the effect.Conclusion:The adenosine A3 receptor is expressed in both the central nervous system pain conduction pathway and the superficial fascia, and participates in the acupuncture analgesia physiological process.
Objective:To investigate the role of adenosine A3 receptor in acupuncture analgesia and explore the expression and distribution of adenosine A3 receptor in the central nervous system pain conduction pathway and the superficial fascia.Methods:Immunohistochemistry and Western blot analysis were used for adenosine A3 receptor in rat L4-S1 spinal segment, brainstem, thalamus, cerebral cortex and after three years, at the midpoint of inguinal and peritoneal fascia expression. Rat tail light dolorimeter was used to analyze the analgesic effect of acupuncture at "Housanli" on thermal radiation pain. Effects of intraperitoneal injection of IB-MECA and Reversine on acupuncture analgesia were tested.Results:Immunohistochemistry and Western blot analysis showed that adenosine A3 receptors were distributed in the spinal cord, brainstem, thalamus and cerebral cortex in the pain transmission pathway, and the highest expression was in the spinal cord, followed by thalamus, cerebral cortex and brainstem. The adenosine A3 receptor was also distributed in "Housanli", the inguinal midpoint and the superficial peritoneum fascia. Western blot analysis showed that the peritoneal expression was higher than that in the groin and "Housanli", but the trend of light density in each group was not obvious by immunohistochemical analysis. After acupuncture at "Housanli" for 5 min, 10 min, 15 min, 20 min, 25 min, 30 min,tail flick latency was significantly prolonged, with time dependence, and needling for 15 min to 20 min had the best analgesic effect. Adenosine A3 receptor agonist IB-MECA enhanced the acupuncture effect while adenosine A3 receptor antagonist reversine can weaken the effect.Conclusion:The adenosine A3 receptor is expressed in both the central nervous system pain conduction pathway and the superficial fascia, and participates in the acupuncture analgesia physiological process.
引文
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[2] Goldman N, Chen M, Fujita T, et al. Adenosine A1 receptors mediate local antinociceptive effects of acupuncture[J]. Nature Neuroscience, 2010, 13(7):883-889.
[3] Horiuchi H, Ogata T, Morino T, et al. Adenosine A1 receptor agonists reduce hyperalgesia after spinal cord injury in rats[J]. Spinal Cord, 2010, 48(9):685-690.
[4] Wang JY, Li H, Zhang L, et al. Adenosine as a probing tool for the mechanistic study of acupuncture[J]. Clinical and Experimental Pharmacology and Physiology, 2014,41(9):933-939.
[5] 陈江旭,郑莉琴,何逸鹏. 腺苷A1与A3受体参与的DRG神经元内Ca2+释放的多光子成像[J]. 光电子·激光, 2014, 25(4):811-817.
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[7] 王子旭, 杨毅, 王卫, 等. 澳大利亚针灸标准化现状分析[J]. 国际中医中药杂志, 2012, 34(5):390-393.
[8] 刘朝, 齐淑兰, 王晓红, 等.《中国针灸》30 年来临床研究类论文质量的调查与思考[J]. 国际中医中药杂志, 2012, 34(5):432-435.
[9] Fang CJ, Yang WJ, Chen YF. Research progress of acupuncture-moxibustion for insomnia:An analysis of literature in recent 5 years[J]. Journal of Acupuncture and Tuina Science, 2013,11(3):181-186.
[10] Sawynok J. Adenosine receptor targets for pain[J]. Neuroscience, 2016,338:1-18.
[11] Sneyd JR, Langton JA, Allan LG,et al. Multicentre evaluation of the adenosine agonist GR79236X in patients with dental pain after third molar extraction[J]. Br J Anaesth, 2007, 98(5):672-676.
[12] Schenone S, Brullo C, Musumeci F,et al. A1 receptors ligands: past, present and future trends[J]. Curr Top Med Chem, 2010,10(9):878-901.
[13] Zylka MJ. Pain-relieving prospects for adenosine receptors and ectonucleotidases[J]. Trends Mol Med, 2011, 17(4):188-196.
[14] 范刚启,钱俐俐,赵杨,等.针刺镇痛机制的多样性及问题分析[J].中国针灸, 2013, 33(1):92-96.
[15] Vase L, Baram S, Takakura N, et al. Specifying the nonspecific components of acupuncture analgesia[J]. Pain, 2013,154(9):1659-1667.
[16] 李忠仁. 实验针灸学[M].北京:中国中医药出版社, 2007:154-231.
[17] 原林, 焦培峰, 唐雷, 等.中医经络理论的物质基础--结缔组织、筋膜和自体监控系统[J]. 中国基础科学, 2005, (3):44-48.
[18] Langevin HM, Churchill DL, Fox JR, et al.Biomechanical response to acupuncture needling in humans[J]. J A ppl Physiol, 2001, 91(6):2471-2478.
[19] 原林, 姚大卫, 唐雷, 等. 针灸经穴的数字解剖学研究[J]. 解剖学报, 2004, 35(4):337-343.
[20] 姜雪梅, 张学全, 原林. 结缔组织在针刺力学信号转导中的作用和研究进展[J]. 针刺研究, 2009, 34(2):136-139.
[2] Goldman N, Chen M, Fujita T, et al. Adenosine A1 receptors mediate local antinociceptive effects of acupuncture[J]. Nature Neuroscience, 2010, 13(7):883-889.
[3] Horiuchi H, Ogata T, Morino T, et al. Adenosine A1 receptor agonists reduce hyperalgesia after spinal cord injury in rats[J]. Spinal Cord, 2010, 48(9):685-690.
[4] Wang JY, Li H, Zhang L, et al. Adenosine as a probing tool for the mechanistic study of acupuncture[J]. Clinical and Experimental Pharmacology and Physiology, 2014,41(9):933-939.
[5] 陈江旭,郑莉琴,何逸鹏. 腺苷A1与A3受体参与的DRG神经元内Ca2+释放的多光子成像[J]. 光电子·激光, 2014, 25(4):811-817.
[6] 李忠仁. 实验针灸学[M]. 北京:中国中医药出版社, 2003:327.
[7] 王子旭, 杨毅, 王卫, 等. 澳大利亚针灸标准化现状分析[J]. 国际中医中药杂志, 2012, 34(5):390-393.
[8] 刘朝, 齐淑兰, 王晓红, 等.《中国针灸》30 年来临床研究类论文质量的调查与思考[J]. 国际中医中药杂志, 2012, 34(5):432-435.
[9] Fang CJ, Yang WJ, Chen YF. Research progress of acupuncture-moxibustion for insomnia:An analysis of literature in recent 5 years[J]. Journal of Acupuncture and Tuina Science, 2013,11(3):181-186.
[10] Sawynok J. Adenosine receptor targets for pain[J]. Neuroscience, 2016,338:1-18.
[11] Sneyd JR, Langton JA, Allan LG,et al. Multicentre evaluation of the adenosine agonist GR79236X in patients with dental pain after third molar extraction[J]. Br J Anaesth, 2007, 98(5):672-676.
[12] Schenone S, Brullo C, Musumeci F,et al. A1 receptors ligands: past, present and future trends[J]. Curr Top Med Chem, 2010,10(9):878-901.
[13] Zylka MJ. Pain-relieving prospects for adenosine receptors and ectonucleotidases[J]. Trends Mol Med, 2011, 17(4):188-196.
[14] 范刚启,钱俐俐,赵杨,等.针刺镇痛机制的多样性及问题分析[J].中国针灸, 2013, 33(1):92-96.
[15] Vase L, Baram S, Takakura N, et al. Specifying the nonspecific components of acupuncture analgesia[J]. Pain, 2013,154(9):1659-1667.
[16] 李忠仁. 实验针灸学[M].北京:中国中医药出版社, 2007:154-231.
[17] 原林, 焦培峰, 唐雷, 等.中医经络理论的物质基础--结缔组织、筋膜和自体监控系统[J]. 中国基础科学, 2005, (3):44-48.
[18] Langevin HM, Churchill DL, Fox JR, et al.Biomechanical response to acupuncture needling in humans[J]. J A ppl Physiol, 2001, 91(6):2471-2478.
[19] 原林, 姚大卫, 唐雷, 等. 针灸经穴的数字解剖学研究[J]. 解剖学报, 2004, 35(4):337-343.
[20] 姜雪梅, 张学全, 原林. 结缔组织在针刺力学信号转导中的作用和研究进展[J]. 针刺研究, 2009, 34(2):136-139.