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KMID : 0363120080210020112
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Korean Journal of Pain
2008 Volume.21 No. 2 p.112 ~ p.118
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A Proteomic Approach for Quantitative Analysis of Calcitonin Gene-related Peptides in the Cerebrospinal Fluid Obtained from a Rat Model of Chronic Neuropathic Pain
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Kim Dong-Hee
Hong Sung-Ho
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Abstract
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Background: This study was conducted to quantitatively analyze proteins associated with the calcitonin gene-related peptide (CGRP) in cerebrospinal fluid (CSF) that was obtained from a rat model of chronic neuropathic pain following administration of intrathecal CGRP8-37.
Methods: Male Sprague-Dawley rats (100?150 g, 5?6 wks) were divided into two groups, sham controls and neuropathic pain models. At the time of operation for neuropathic pain model, an intrathecal catheter was threaded through the intrathecal space. At 1 or 2 wks after the operation (maximum pain state), a test dose of 1, 5, 10, or 50 nM of CGRP8-37 was injected into the intrathecal catheter and the CSF was then aspirated. Conventional proteomics to evaluate the CSF were then performed using high resolution 2-D, gel electrophoresis followed by computational image analysis and protein identification by mass spectrometry.
Results: Treatment with CGRP8-37 effectively alleviated mechanical allodynia in a dose dependent manner. The most effective response was obtained when a dose of 50 nM was administered, but significant differences were obtained following administration of only 5 nM CGRP8-37. Furthermore, the results of the proteomic analysis were consistent with the experimental results. Specially we detected 30 differentially expressed spots in 7 images when 2-D gel electrophoresis was conducted. The intensity of 6 of these spots (spot number: 20 and 26?30) was found decrease the CGRP8-37 dose increased; therefore, these spots were evaluated by mass spectrometry. This analysis identified 2 different proteins, CGRP (spot numbers: 26?30) and neurotensin-related peptide (spot
number: 20).
Conclusions: The results of this study suggest that CGRP plays a role in chronic central neuropathic pain and is a major target of chronic neuropathic pain management.
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KEYWORD
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allodynia, CGRP, electrophoresis, neuropathic pain, NRP, proteomics
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