论文标题:白细胞介素-2对急性脊髓损伤细胞凋亡的影响相关性实验研究
The Effect of Interleukin-2 on Apoptosis Following Spinal Cord Injury in Rats
论文作者 王颖
论文导师 陈晓亮,论文学位 硕士,论文专业 外科学
论文单位 青岛大学,点击次数 522,论文页数 52页File Size4117k
2005-03-15论文网 http://www.lw23.com/lunwen_1002522/ 脊髓损伤;白细胞介素-2;细胞凋亡
spinal cord injury; interleukin-2; apoptosis
目的:在建立大鼠脊髓损伤模型基础上,观察不同时间点IL—2对脊髓胶质细胞,神经元凋亡的影响,以期为临床急性脊髓损伤的治疗提供理论依据。 方法:健康成年大鼠72只,体重250～300g,随机分为两组,其中单纯损伤组(单纯打击损伤)36只,IL—2组(手术加IL—2)36只。脊髓损伤动物模型的制备:用自制Allen打击器25gcm致伤力撞击脊髓,制成脊髓损伤动物模型。IL—2组每日肌注IL—2(10u)直至处死,单纯损伤组仍每日肌注生理盐水0.5ml直至处死。组织切片的制备:脊髓损伤后1d,3d,5d,8d,14d,21d共6个时间段分批处死动物。4%多聚甲醛心肌灌注固定24h,切取每只大鼠T_5～T_(13)节段脊髓组织,常规制成切片,给于HE染色,Bcl-2检测,TUNEL原位末端标记以及伤后第14天检测Gale评分和斜板实验,检测大鼠脊髓损伤后细胞凋亡和脊髓神经功能恢复情况。 结果:TUNEL原位标记检测:单纯损伤组24小时已出现不少阳性细胞,以胶质细胞为主,3—8天达到高峰,此后逐渐回落,但21天时仍有阳性细胞,应用IL—2治疗后,凋亡细胞明显减少(P<0.05)。Bcl-2蛋白检测:在脊髓损伤后的第1天,脊髓组织即开始较高表达Bcl-2蛋白。单纯损伤组Bcl—2高峰发生在损伤后3天,而IL—2组伤后8天达到高峰,此时Bcl—2蛋白不仅表达在神经细胞中,更多的在胶质细胞中大量表达,且此状态一直维持到伤后14天才开始下降,伤后21天仅少量表达(P<0.05)。伤后第14天检测Gale评分和斜板实验,IL—2组同单纯损伤组相比,无明显区别(P>0.05)。 结论:本研究证明,IL—2在脊髓损伤中通过改善微循环,促进Bcl-2蛋白的表达,清除氧自由基,抗氧化作用能够抑制脊髓损伤早期神经细胞和胶质细胞的凋亡,从而减轻脊髓继发性损伤,促进脊髓神经功能的恢复。所以SCI后早期应用IL—2配合其它药物阻止神经细胞和胶质细胞的凋亡可能有助于减轻SCI的损害程度,促进脊髓神经功能的恢复,为SCI的治疗提供一个新的途径。
Objective: In rats model of spinal cord injury,to explore the interleukin-2 on apoptosis followingspinal cord injury in different time.Methods: 72 healthy rats were chosen and their body weight varied from 250 gram to 310 gram.The rats were divided into two groups randomly:controlled group(36) and interleukin-2 group(36). All of rats spinal cord injury model were established with ALLEN bump equipment(25gcm).The model of IL-2 group injected IL-2(10u) every day until they were sacrificed. The model of controlled group injected NS(0.5ml/d) every day until they were sacrificed. Fabrication of tissue section:after spinal cord injury ld,3d,5d,8d,14d,21d,we sacrificed rats in batch.4% formaldehydum polymerisatum were pured into the cardiac muscle and fixed for 24h.Then obtain the specimen of spinal cord from T_5 to T_(13).Establishing slices,HE dyeing,bcl-2 dectation,terminal doxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL).On the fourteenth day after spinal cord injury,we detect Gale grading score and tiltboard experiment. Result: Tunel dectation:In control group,after 24 hours,there are many positive cells,most of them are gliao cells,it reached the peak after 3-8 day,and then,they were decreased gradually.in IL-2 group,positive cells are obviously less than those of control group(P<0.05).Bcl-2 dectation:During the first day after spinal cord injury,we find many bcl-2 protein in the spinal cord tissue.In control group,they reached the peak on the third day after spinal cord injury. In IL-2 group,they reached the peak on the eighth day after spinal cord injury.Bcl-2 protein are more in glial cells than those in nerve cells.They begin to decrease after the fourteenth day of spinal cord injury (P<0.05) . The chang of hindlimb motor function on the fourteenth day after spinal cord injury: IL-2 group is not better than control group (P>0.05) .Conclusion: From the investigate,we proved that IL-2 can improve the microcirculation,get rid of oxygen free radical,resist oxygenation,so as to restrain the apoptosis of nerve cell and glial cell,lesson spinal cord injury,improve to recover spinal nerve.Therefore,during early time of spnal cord injury,we make use of IL-2 with other medicine,it can lesson spinal cord injury and provide a new method for treatment of spinal cord injury.
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