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张 杨,王永慧,丁欣利,王艳琴,王 荣,岳寿伟.TRPV4在介导大鼠背根神经节持续受压后机械和热痛敏中的作用[J].中国康复医学杂志,2010,25(12):1123~1130
TRPV4在介导大鼠背根神经节持续受压后机械和热痛敏中的作用    点此下载全文
张 杨  王永慧  丁欣利  王艳琴  王 荣  岳寿伟
山东大学齐鲁医院康复科,济南,250012
基金项目:国家自然科学基金资助课题(30872732,81071597);山东省自然科学基金资助课题(Z2007C04)
DOI:
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摘要:
      摘要 目的:观察持续机械压迫(CCD)对瞬时感受器电位离子通道4(TRPV4)基因、蛋白表达及功能的影响,明确TRPV4是否参与CCD导致的机械和热痛敏。 方法:建立CCD模型后,分别于手术前及手术后第7天、第14天及第28天取材前测量运动功能、机械刺激缩爪反应阈值和热辐射刺激缩爪反应潜伏期。为了测量TRPV4反义核苷酸干扰对机械和热痛阈值的影响,在蛛网膜下腔内注入TRPV4寡脱氧核苷酸(ODN) 40μg/d,每天1次,第7天后测量大鼠行为学变化。使用实时定量RT-PCR检测TRPV4基因表达的变化,Western blot检测TRPV4蛋白质表达量的变化,激光共聚焦检测低渗溶液和佛波醇(4?琢-PDD)刺激背根神经节(DRG)神经元后细胞内钙离子浓度的变化。 结果:所有动物在损伤前后步态均正常,持续压迫明显降低大鼠的机械和热痛阈,TRPV4干扰可部分逆转该痛敏。持续机械压迫可以明显增加TRPV4基因和蛋白的表达,手术后第7天,第14天和第28天,TRPV4 mRNA的表达分别为假手术组大鼠的4.29倍、2.95倍和2.48倍, 蛋白表达量分别为假手术组大鼠的4.34倍,3.88倍和2.47倍。持续机械压迫后,对低渗溶液和4?琢-PDD产生反应的DRG神经元的比例数增加,细胞内钙的峰值增高。这种反应被TRPV4反义ODN所抑制。 结论:CCD可以上调TRPV4的基因、蛋白表达,敏化通道的功能;TRPV4参与介导CCD导致的机械和热痛敏。
关键词:机械性痛敏  热痛敏  瞬时感受器电位离子通道4  背根神经节  持续压迫
The role of transient receptor potential vanilloid 4 in mediating mechanical and thermal allodynia following chronic compression of dorsal root ganglion in rats    Download Fulltext
Department of Physical Medicine and Rehabilitation, Qilu Hospital of Shandong University, Jinan, 250012
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Abstract:
      Abstract Objective: To investigate the role of transient receptor potential vanilloid 4(TRPV4) in mediating mechanical and thermal allodynia in rodent models of chronic compression of dorsal root ganglion (CCD). Method: The levels of TRPV4 mRNA and protein expressions in dorsal root ganglion (DRG) were assessed using real-time RT-PCR and Western blot analysis respectively at the 7th, 14th and 28th d post-CCD. The effects of spinal administration of TRPV4 antisense oligodeoxynucleotide(ODN) and mismatch ODN on CCD-induced mechanical and thermal allodynia were evaluated. The calcium responses to hypotonic solution and 4α-phorbol 12, 13-didecanoate (4α-PDD) were assessed following sham surgery, CCD, spinal application of TRPV4 antisense ODN and mismatch ODN. Result: The levels of TRPV4 mRNA and protein expression increased significantly at the 7th-28th d post-CCD with the highest level at the 7th d post-CCD when compared with the sham group. TRPV4 antisense ODN, but not mismatch ODN, partly reversed the CCD-induced mechanical and thermal allodynia. Additionally, TRPV4 antisense ODN had no effect on the baseline nociceptive response. The percentage of DRG neurons responsive to hypotonic solution and 4α-PDD and the fluorescence ratio of calcium response also enhanced significantly in both CCD group and mismatch ODN group. These increased responses were significantly inhibited by TRPV4 antisense ODN. Conclusion: TRPV4 plays a crucial role in CCD-induced mechanical and thermal allodynia.
Keywords:mechanical allodynia  thermal allodynia  transient receptor potential vanilloid 4  dorsal root ganglion  chronic compression
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