CAJ | 학술논문

脊髓损伤是一类常见的、高致残率的中枢神经系统疾病,由于多种复杂因素影响其损伤后的修复过程,损伤脊髓的再生能力非常有限.本研究采用cDNA微阵列技术筛选大鼠脊髓损伤后出现的差异表达基因.实验组动物在T8-T9进行脊髓全横断手术,对照组动物只打开椎板;4.5 d后取脊髓进行RNA提取并在反转录过程中进行Cy3/Cy5标记,然后与预制的、带有4 041条特异性探针的芯片进行杂交.Cy5/Cy3信号比值≥2.0视为脊髓损伤后出现差异表达的基因.通过筛选,我们得到了65个上调表达基因(21个已知基因,30个已知EST和14个未知基因)和79个下调基因(20个已知基因,42个已知EST和17个未知基因).进一步通过半定量RT-PCR对其中的5个上调已知基因(Timp1,Tagln,Vim,Fc gamma receptor.Ctss)和三个下调已知基因(stearyl-CoA desaturase,F2,Ensa)的表达情况进行了验证,结果显示与芯片结果一致.这些基因可能在脊髓损伤后的修复过程中起一定的作用,对其深入研究将有助于揭示脊髓损伤修复的分子机制.
척수손상시일류상견적、고치잔솔적중추신경계통질병,유우다충복잡인소영향기손상후적수복과정,손상척수적재생능력비상유한.본연구채용cDNA미진렬기술사선대서척수손상후출현적차이표체기인.실험조동물재T8-T9진행척수전횡단수술,대조조동물지타개추판;4.5 d후취척수진행RNA제취병재반전록과정중진행Cy3/Cy5표기,연후여예제적、대유4 041조특이성탐침적심편진행잡교.Cy5/Cy3신호비치≥2.0시위척수손상후출현차이표체적기인.통과사선,아문득도료65개상조표체기인(21개이지기인,30개이지EST화14개미지기인)화79개하조기인(20개이지기인,42개이지EST화17개미지기인).진일보통과반정량RT-PCR대기중적5개상조이지기인(Timp1,Tagln,Vim,Fc gamma receptor.Ctss)화삼개하조이지기인(stearyl-CoA desaturase,F2,Ensa)적표체정황진행료험증,결과현시여심편결과일치.저사기인가능재척수손상후적수복과정중기일정적작용,대기심입연구장유조우게시척수손상수복적분자궤제.
The acute traumatic spinal cord injury (SCI) is a commonly seen and severe case in clinic. However, the repair and regeneration of injured spinal cord is limited. This is likely due to that different kinds of factors are involved in regeneration after SCI.In the present study, we used complementary DNA microarray consisting of 4 041 specific probes from rat to identify genes that were differentially expressed after SCI. The animals were subjected to complete transection injury of the thoracic spinal cord (T8-T9). Sham operated animals Receivedonly a laminectomy. Four and a half days later, rat spinal cord was dissected out for total RNA isolation. The fluorescent (Cy3 and Cy5) labeled probes were prepared and hybridized to the microarray. Genes that showed 2-fold difference in SCI tissue were identified. Sixty-five up-regulated genes consisted of 21 known genes, 30 known expressed sequence tags (ESTs) and 14unknown genes. Seventy-nine down-regulated genes comprised 20 known genes, 42 known ESTs and 17 unknown genes. In 41differentially expressed known genes, 5 up-regulated genes, i.e., tissue inhibitor of metalloproteinase 1 (Timpl), transgelin (Tagln),vimentin (Vin), Fc gamma receptor, cathepsin S (Ctss), and 3 down-regulated genes, i.e., stearyl-CoA desaturase, coagulation factor H (F2), endosulfin alpha (Ensa), were further confirmed by reverse transcription polymerase chain reaction (RT-PCR). These genes may play a role in the response to tissue damage or repair following SCI and characterization of them might be helpful to elucidate the molecular mechanisms of spinal cord injury and regeneration.