CAJ | 학술논문

背景:Cathepsis 家族是否参与脊髓损伤早期的病理过程以及大剂量甲基强的松龙是否通过溶酶体机制发挥神经保护作用目前尚不清楚.目的:检测 Cathepsin 基因家族在脊髓损伤早期的表达和大剂量甲基强的松龙干预后的变化,明确大剂量甲基强的松龙是否通过调节溶酶体凋亡途径发挥神经保护作用.方法:9只日本大耳兔随机分为3组:模型组和药物组进行椎板切除后采用Al en法建立急性脊髓损伤模型,药物组在造模后2 h按人-兔等效剂量给予大剂量甲基强的松龙冲击治疗,对照组仅进行椎板切除.造模后8 h 处死实验动物,取脊髓组织,采用 Trizol法提取总 RNA,用9张 Agilent 兔子全基因4*44K 芯片进行检测.采用 GeneSpring 10.0软件,以 P <0.05且倍数变化(FC)≥2筛选出差异表达基因.结果与结论:成功建立脊髓损伤的动物模型并获得相应的组织标本.9组标本总 RNA 的质量均能满足基因芯片检测要求.基因芯片结果显示:在10个Cathepsin 基因家族成员中,仅Cathepsin Z 和 proathepsin E 在创伤后呈现差异性表达,Cathepsin C、D、F、K、L、S 和 W 表达均无差异.甲基强的松龙冲击治疗后 Cathepsin 家族基因表达均无差异(在药物组与模型组的比较).提示 Cathepsin Z 和 E 参与了脊髓损伤早期凋亡过程,但大剂量甲基强的松龙并不能通过溶酶体凋亡途径发挥神经保护作用.
배경:Cathepsis 가족시부삼여척수손상조기적병리과정이급대제량갑기강적송룡시부통과용매체궤제발휘신경보호작용목전상불청초.목적:검측 Cathepsin 기인가족재척수손상조기적표체화대제량갑기강적송룡간예후적변화,명학대제량갑기강적송룡시부통과조절용매체조망도경발휘신경보호작용.방법:9지일본대이토수궤분위3조:모형조화약물조진행추판절제후채용Al en법건립급성척수손상모형,약물조재조모후2 h안인-토등효제량급여대제량갑기강적송룡충격치료,대조조부진행추판절제.조모후8 h 처사실험동물,취척수조직,채용 Trizol법제취총 RNA,용9장 Agilent 토자전기인4*44K 심편진행검측.채용 GeneSpring 10.0연건,이 P <0.05차배수변화(FC)≥2사선출차이표체기인.결과여결론:성공건립척수손상적동물모형병획득상응적조직표본.9조표본총 RNA 적질량균능만족기인심편검측요구.기인심편결과현시:재10개Cathepsin 기인가족성원중,부Cathepsin Z 화 proathepsin E 재창상후정현차이성표체,Cathepsin C、D、F、K、L、S 화 W 표체균무차이.갑기강적송룡충격치료후 Cathepsin 가족기인표체균무차이(재약물조여모형조적비교).제시 Cathepsin Z 화 E 삼여료척수손상조기조망과정,단대제량갑기강적송룡병불능통과용매체조망도경발휘 신경보호작용.
BACKGROUND: It is not clear whether the Cathepsis family is involved in the pathological process of early spinal cord injury and whether high-dose methylprednisolone plays neuroprotective effect through lysosome apoptosis pathway. OBJECTIVE: To explore the expression and change of genes Cathepsin family in early spinal cord injury and to identify if high-dose methylprednisolone plays neuroprotective effect by the lysosome apoptosis pathway. METHODS: Nine Japanese rabbits were randomly divided into three groups, the rabbits in the model group and drug treatment group were treated with laminectomy, and then the rabbits were used to establish the acute spinal cord injury model with Al en fal ing strike method. The rabbits in the drug treatment group were treated with human equivalent dose flushing-dose methylprednisolone at 2 hours after acute spinal cord injury. The rabbits in the control group were treated with laminectomy. Al rabbits were kil ed at 8 hours after acute spinal cord injury, and then the damaged spinal cord tissues were obtained careful y. Total RNA was extracted from above nine samples with Trizol One-step method to undergo the examination of the gene expression profile by using Agilent Rabbit Oligo Microarray (4×44K) respectively. GeneSpring 11.0 software was then used to filter potential candidate genes, and only genes with P values ≤0.01 and fold change≥2 were retained for further analysis. RESULTS AND CONCLUSION: The spinal cord injury models were successful y set up and the corresponding tissue specimens were obtained. Acquired nine subsample of total RNA were qualified for microarray examination. The results of microarray examination showed that among the 10 genes of Cathepsin family, only Cathepsin Z and Procathepsin E showed significant different expression. Al of Cathepsin family genes of Cathepsin C, D, F, K, L, S and W did not showed significant different expression. There were no significant differences of Cathepsin family genes expressions between drug treatment group and model group after treated with methylprednisolone. Gene Cathepsin Z and Procathepsin E took part in the apoptosis of spinal cord injury at acute phase, but high-dose methylprednisolone cannot play neuroprotective effect by the lysosome apoptosis pathway.