Identification of Genes Expression in Spinal Cord after Peripheral Inflammation Using DNA Microarray Techniques

以DNA微陣列技術辨識周邊組織發炎後之脊髓基因表現

孫孝倫(Hsiao-Lun Sun) ； 鄭宇哲(Yu-Che Cheng) ； 郭書麟(Shu-Lin Guo) ； 曾劍英(Jann-Inn Tzeng) ； 簡志誠(Chih-Cheng Chien) ； 錢良慧(Liang-Hui Chien) ； 凌慶東(Qing-Dong Ling)

《疼痛醫學雜誌》 18卷1期 (2008/03) Pp. 15-24

https://doi.org/10.29792/TTJP.200803.0003

摘要

周邊組織發炎可以暫時性激發許多脊髓背角神經元的基因表現，然而，不少研究指出，在周邊組織發炎後的脊髓背角神經元中，被激發的不只是基因。在本研究中，我們應用DNA微陣列技術辨識周邊組織發炎後之脊髓基因表現。
我們以公SD大鼠做實驗，在其左後腳掌注射complete Freund's adiuvant (CFA) (1:1, CFA: saline, 200μ 1)。24小時後取出其第4-5節腰椎，萃取出RNA後製作cDNA及cRNA，並應用CodeLink大鼠全基因體微陣列晶片，輔以RT-PCR做確認。我們的結果顯示，62個基因表現增加而48個基因表現減少，其中各有32/16個EST／未知基因。我們的結論是：周邊組織發炎可以動態陸的激發許多與疼痛覺相關的脊髓背角神經元基因表現。

關鍵字

脊髓 ； 疼痛覺 ； 基因表現 ； 微陣列

並列摘要

Peripheral inflammation may result in the transcriptional activation of many genes in spinal cord dorsal horn neurons. However, reports of many studies suggested that more than genes should be involved in spinal nociceptive processing after peripheral inflammatory stimulation. In this study, we used microarray techniques to identify genes that were altered in spinal cord after peripheral inflammation. Male rats received an injection of complete Freund's adiuvant (CFA) (1:1, CFA: saline, 200 μ1) into the plantar surface of the left hindpaw. 24hrs after injection, L4-L5 of spinal cord segments were removed and tissues were cut along the midline into ipsilateral and contralateral sides. Total RNA was isolated and synthesis of eDNA and cRNA were carried out. 10 μ g of ipsilateral side (n=4) or contralateral side (n=2) cRNA were hybrid with each chamber of a CodeLink Rat whole genome microarray chip, which contains approximately 33,000 oligonucleotide gene probes. The changes in gene identified through the microarrays analysis were confirmed with quantitative RT-PCR. We screened out total 110 genes that were altered over 2-fold in ipsilateral side comparing contralateral side. Among them, 62 genes including 32 of EST/unknown genes were shown increase and 48 genes including 16 of EST/unknown genes were shown decrease in level of expression. Our results suggest that peripheral inflammation induced the dynamic activation of neuron markers in nociceptive processing of spinal neurons.

並列關鍵字

spinal cord ； nocicepte ； gene expression ； microarray