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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Allen法建立急性脊髓损伤模型,药物组在造模后2h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8h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。
BACKGROUND: Whether the Cathepsis family participates in the early pathological process of spinal cord injury and whether neuroprotection by high-dose methylprednisolone via the lysosomal mechanism is unclear. OBJECTIVE: To detect the expression of Cathepsin gene family in the early stage of spinal cord injury and the changes after intervention with high-dose methylprednisolone, and to determine whether high-dose methylprednisolone can play a neuroprotective role through the regulation of lysosomal apoptotic pathway. Methods: Nine Japanese white rabbits were randomly divided into three groups: acute spinal cord injury model was established by Allen method in model group and drug group after laminectomy. The drug group was given high dose Methylprednisolone impact treatment, the control group only for laminectomy. The experimental animals were sacrificed at 8h after modeling, and the spinal cord tissue was taken. The total RNA was extracted by Trizol method and detected by 9 * 4K gene chips of Agilent rabbit. GeneSpring 10.0 software was used to screen differentially expressed genes with P <0.05 and fold change (FC)> 2. RESULTS AND CONCLUSION: Animal models of spinal cord injury were successfully established and the corresponding tissue specimens were obtained. The quality of total RNA of 9 groups of samples can meet the requirements of gene chip detection. Gene chip results showed that only Cathepsin Z and proathepsin E were differentially expressed after trauma in 10 Cathepsin gene family members, and there was no difference in Cathepsin C, D, F, K, L, S and W expression. There was no difference in Cathepsin family gene expression after methylprednisolone treatment (comparison between drug group and model group). These results suggest that Cathepsin Z and E are involved in the early apoptotic process of spinal cord injury. However, high doses of methylprednisolone do not play a neuroprotective role through the lysosomal apoptotic pathway.