【作者】 沈雄杰；

【导师】 王锡阳；

【作者基本信息】 中南大学 ， 外科学， 2011， 硕士

【摘要】 目的：利用改良Allen’s打击法建立大鼠急性脊髓损伤模型,观察英夫利昔单抗、甲基强的松龙及英夫利昔单抗联合甲基强的松龙对大鼠急性脊髓损伤的治疗疗效,分析英夫利昔单抗、甲基强的松龙及英夫利昔单抗联合甲基强的松龙对损伤段脊髓组织中Bcl-2、Survivin的影响。探讨英夫利昔单抗联合甲基强的松龙对急性脊髓损伤的保护作用及可能机制。材料和方法：90只正常成年SD大鼠,同一批次,雄性,体重控制在300-350g,随机分为6组,正常对照组(Group NC, n=15),假手术组(Group SO, n=15),安慰剂治疗组(Group NS, n=15),甲基强的松龙治疗组(Group MP, n=15),英夫利昔单抗治疗组(Group IN,n=15),英夫利昔单抗联合甲基强的松龙治疗组(Group IM, n=15)。除正常对照组不做手术及治疗处理、假手术组仅作椎板切除外,其余各组均建立急性脊髓损伤(ASCI)模型,并给予相应的药物治疗(甲基强的松龙30mg/kg,腹腔注射；英夫利昔单抗5mg/kg,腹腔注射；单药治疗组给药一次,药物联合治疗组给药两次)。采用改良Allen’s重物打击法制作胸10水平急性脊髓中度损伤模型。急性脊髓损伤(ASCI)模型建立成功后,各实验组立即给予相应的药物治疗,安慰剂治疗组经腹腔给予注射用生理盐水2ml,甲基强的松龙治疗组给予甲基强的松龙(30mg/kg,腹腔注射,注射剂量为2ml,)一次,英夫利昔单抗治疗组(5mg/kg,腹腔注射,注射剂量为2ml)给予英夫利昔单抗一次,英夫利昔单抗联合甲基强的松龙治疗组给予英夫利昔单抗及甲基强的松龙各一次(英夫利昔单抗,5mg/kg;甲基强的松龙,30mg/kg;腹腔注射；分两次给药),正常对照组不予处理,假手术组于相应时间经腹腔给予注射用生理盐水lml。每组取5只大鼠对其进行BBB评分,剩余大鼠于脊髓损伤后第24h处死并获取脊髓标本,检测指标包括：1.使用免疫组化分析Bcl-2、Survivin表达情况。2.使用RT-PCR分析Bcl-2 mRNA、Survivin mRNA表达情况结果：(1)BBB评分：在伤后14、21天,MP治疗组和Infliximab治疗组的大鼠BBB评分大于安慰剂治疗(p<0.05),同时MP联合Infliximab治疗组的大鼠BBB评分又大于MP和Infliximab单独治疗组(p<0.05)。(2)免疫组化：在伤后24h,MP治疗组和Infliximab治疗组表达的Bcl-2和Survivin大于安慰剂治疗(p<0.05),同时MP联合Infliximab治疗组表达的Bcl-2和Survivin又大于MP和Infliximab单独治疗组(p<0.05)。(3)RT-PCR：在伤后24h,MP治疗组和Infliximab治疗组表达的Bcl-2 mRNA和Survivin mRNA大于安慰剂治疗(p<0.05),同时MP联合Infliximab治疗组表达的Bcl-2 mRNA和Survivin mRNA又大于MP和Infliximab单独治疗组(p<0.05)。结论：(1)英夫利昔单抗、甲基强的松龙对大鼠脊髓损伤有效,二者联用效果更佳。(2)英夫利昔单抗、甲基强的松龙均可通过促进抗凋亡蛋白Bcl-2、Survivin的表达减轻脊髓的损伤程度及改善脊髓损伤后神经功能障碍。更多还原

【Abstract】 Objective:By the modified Allen’s weight drop method, a model of acute spinal cord injury(ASCI) in rats was framed. Observation of infliximab, methylprednisolone and infliximab combined methylpredni-solone on acute spinal cord injury therapy, analyzed of Infliximab, methylprednisolone and infliximab combined methylprednisolone impact on Bcl-2, Survivin, expression of injured spinal cord tissues. To investigate the protective effect and possible mechanism of infliximab combined methylprednisolone.Materials and methods:90 adult Sprague Dawley rats with 300-350g body weight(BWT)were randomly divided into six primay groups:(1)Normal control group (group NC n=15);(2)Sham operation group (group SO n=15);(3) Placebo group (group NS n=15);(4) Methylprednisolone treated group(group MP N=15);(5) Infliximab treated group(group IN n=15);(6) infliximab combined methylprednisolone treated group(group IM n=15).Excepted non-surgical and non-treatment in group NC,only cut lamina in group SO,other groups were established ASCI model, and appropriate drug treated.(Methylprednisolone 30mg/kg, intraperitoneal injection; infliximab 5mg/kg, intraperitoneally; monotherapy group administered once, twice administration of drug combination therapy).Allen’s weight drop method was used to establish the acute spinal cord injury (ASCI)rat model at T10 section.The appropriate drug treatment given at once, after the model was established.Group NS given normal saline 2ml by intraperitoneal injection, Group MP given methylprednisolone once (30mg/kg, intraperitoneal injection, injecting a dose of 2ml),Group IN given infliximab once (5mg/kg, intraperitoneal injection, injecting a dose of 2ml),Group IM given methylprednisolone infliximab and methylpred-nisolone(infliximab 5mg/kg, methylprednisolone 30mg/kg, intraperitoneal injection, divided doses).Group NC not to processing. Group SO given by intraperitoneal injection of nonnal saline lml in corresponding time.5 rats in each group were evaluton by BBB(Basso,Beattie, Bresnahan) locomotor rating, the remaining rats were killed 24h after spinal cord injury and spinal cord specimens were obtained. Detection indicators include:1 By immunohistochemical analysis Bcl-2, survivin expression; 2 By RT-PCR analysis Bcl-2 mRNA、survivin mRNA expression.Result:1 After injury 14d,21d, the BBB scores of rats in the group MP and the group IN were greater than the group NS (p<0.05), while MP combined Infliximab treatment group rats BBB score is greater than MP or Infliximab monotherapy groups (p<0.05). After injury 24h, MP or Infliximab treatment group treated group greater than placebo group in the expression of Bcl-2 and Survivin (p<0.05), while MP combined Infliximab treatment group was greater than MP or Infliximab monotherapy group in the expression of Bcl-2 and Survivin (p<0.05) After injury 24h, MP or Infliximab treatment group treated group greater than placebo group in the expression of Bcl-2 mRNA and Survivin mRNA (p<0.05), while MP combined Infliximab treatment group was greater than MP or Infliximab monotherapy group in the expression of Bcl-2 mRNA and Survivin mRNA (p<0.05)Conclusions:(1) infliximab, methylprednisolone on spinal cord injury has therapeutic effect, combined them with better effect. (2) infliximab, methylprednisolone can be through the promotion of anti-apoptotic protein Bcl-2, Survivin expression and reduce the degree of spinal cord injury to improve neurological function after spinal cord injury.更多还原