MP对大鼠（环扎法）急性脊髓损伤后HSP70的表达的影响

发布时间：2018-05-02 17:12

本文选题：脊髓损伤(SCI) + 动物模型　；参考：《皖南医学院》2015年硕士论文

【摘要】：目的:通过对随机选入的SD大鼠采用环扎法制造大鼠脊髓损伤的模型,研究脊髓损伤后MP的使用与HSP70的表达变化规律,探究初期脊髓损伤的医治策略,为进一步学习脊髓损伤的模型打下坚实的理论基础。方法:本课题在前期工作基础上再次通过大鼠环扎法造成大鼠脊髓损伤的模型,该方法具有很强的重复性,且方法简单稳定,适合研究本课题。随机选入成年SD雄性大鼠60只,大小均为13周龄,分为三组,第一组为空白组即A组:仅行椎板减压,不作脊髓环扎处理。第二组为对照组即B组:采用环扎法进行脊髓环扎,术后6小时作脊髓减压处理;试验治疗组为C组:同B组方法一致,也行脊髓环扎处理,在环扎法术后6小时减压,但在环扎术后1小时给予大剂量MP静脉注射治疗。环扎术的关键就是充分地将研究段脊髓清晰暴露,本方法以蚊式血管钳在肉眼情况下逐步咬除胸11椎板,暴露出脊髓,使用POLYSORB 5-0白色缝合线,精确地测量出硬膜囊周长(C1),取出缝合线,通过公式(C2=C1×7.0)进行运算,得出需要的C2值,也就是硬膜囊原横截面积的70%,通过可吸收线将原测量平面上的脊髓环扎至70%。之后分别于脊髓环扎术后第一天、第三天、第七天、第十四天和第二十一天等时间段对各组大鼠分批进行处死,并取各组大鼠受损脊髓进行制作病理切片,选用HE染色、免疫组化及原位末端脱氧核糖核酸转移酶介导的脱氧尿苷三磷酸(dUTP)标记法(TUNEL法)检查和采用双人盲法结合BBB评分、斜板法评价对各组结果进行统计学分析,做出相关性研究。结果:空白组A组神经运动功能BBB评分、斜板评分、HE染色均未见明显异常改变;大鼠脊髓组织中HSP70阳性细胞数比较少见。与空白组A组相比,对照组B组大鼠的神经运动功能BBB评分、斜板评分均降低,切片观察脊髓组织中HSP70阳性细胞数于术后第1天开始升高,术后第3天达到最大值,差别有统计学意义(每组之间数据比较P0.05)。与对照组B组比较,治疗组C组大鼠的神经运动功能BBB评分、斜板评分明显增高,脊髓组织中HSP70阳性细胞数明显增加,HSP70阳性细胞数于术后第3天达到峰值,差异有统计学意义(各组之间数据比较P0.05)。结论:1、环扎法是通过缝合线对脊髓进行持续性静态压迫而制作出大鼠急性脊髓损伤的模型,该方法的重复性良好,可定量化及可结合临床,是一种比较可靠的脊髓损伤造模方法。2、实验组与对照组之间的BBB评分、斜板评分、HSP70阳性细胞数均存在统计学差异,由此可以推论MP可增加大鼠脊髓损伤后HSP70的表达,而HSP70的作用机制为改善急性脊髓损伤后钙离子的不平衡,抑制脂质过氧化反应,避免自由基的损害,还能够增加前列腺素的合成,改善损伤脊髓的血运状况,加强脊髓组织的修复,并减少了脊髓损伤后继发性损害的发生,起到了保护神经的作用。3、脊髓损伤后,在外界因素的应激作用下,机体内HSP70表达增高,研究发现该蛋白为保护性蛋白,有保护机体脊髓神经的功效,避免脊髓的进一步损伤,对后期机体脊髓功能的恢复有一定的作用。
[Abstract]:Objective: To study the model of rat spinal cord injury by ligation of random selected SD rats, to study the changes in the use of MP and the expression of HSP70 after spinal cord injury, to explore the treatment strategy of spinal cord injury in the early stage, and to lay a solid theoretical basis for further learning of the model of spinal cord injury. The rat model of spinal cord injury caused by rat ring ligature was repeated, and the method was very repeatable, and the method was simple and stable. 60 adult SD male rats were randomly selected, the size of which was 13 weeks old and divided into three groups. The first group was the blank group, A group, only the vertebral lamina decompression and no spinal cord ligature. The second group was the same. Group B: group B: spinal cord ligature and decompression of spinal cord after operation. The test group was group C: same as group B, spinal cord cerclage, 6 hours after cerclage, 1 hours after cerclage, with large dose of MP intravenous injection. The key of cerclage was to fully study the spinal cord. In this method, the 11 vertebral lamina was gradually bitten by the mosquito type vascular forceps in the naked eye, the spinal cord was exposed, the POLYSORB 5-0 white suture was used, the perid sac circumference (C1) was accurately measured, the suture was taken out, the C2 value needed was obtained through the formula (C2=C1 x 7), that is, 70% of the original cross section of the dural sac, through the absorbable line. The spinal cord in the original measurement plane was ligation to 70%. after the first day, third days, seventh days, fourteenth days and twenty-first days after the spinal cord cerclage, and the injured spinal cord of each group was taken to make pathological sections, and HE staining, immunohistochemistry and in situ DDT were used. Transfer enzyme mediated deoxy uridine three phosphoric acid (dUTP) labelling (TUNEL) examination and double blind method combined with BBB score. The results of each group were statistically analyzed by oblique plate method, and the correlation study was made. Results: there was no obvious abnormal change in the BBB score of nerve movement function in the blank group A group, the score of oblique plate, HE staining, and the spinal cord tissue of rats. The number of HSP70 positive cells was rare. Compared with the A group in the blank group, the neuromotor function BBB score of the control group of the control group and the score of the oblique plate decreased. The number of HSP70 positive cells in the spinal cord tissue began to rise at first days after the operation, and the maximum value was reached on the third day after the operation, and the difference has the significance of the statistical significance (the comparison of the data between each group, P0.05). And the control. Group B was compared with the BBB score of the neuromotor function in the C group of the treatment group, the score of the oblique plate increased obviously, the number of HSP70 positive cells in the spinal cord increased obviously, and the number of HSP70 positive cells reached the peak at third days after the operation. The difference was statistically significant (the comparison of the data between each group was P0.05). Conclusion: the ring ligature method is the continuation of the spinal cord through the suture line to the spinal cord. The model of acute spinal cord injury in rats is made by sexual static compression. The reproducibility of this method is good. It can be quantified and can be combined with clinical. It is a reliable method of making model.2 for spinal cord injury. The BBB score, the score of oblique plate and the number of HSP70 positive cells between the experimental group and the control group are statistically different, thus the inference that the MP can be increased can be deduced. The expression of HSP70 after spinal cord injury in rats, and the action mechanism of HSP70 is to improve the imbalance of calcium ion after acute spinal cord injury, inhibit lipid peroxidation, avoid the damage of free radicals, increase the synthesis of prostaglandins, improve the blood transport status of the injured spinal cord, strengthen the repair of spinal cord tissue, and reduce the secondary sex of spinal cord injury. The occurrence of damage has played a role in protecting the nerve.3. After the injury of the spinal cord, the expression of HSP70 in the body is increased under the stress of external factors. The study found that the protein is a protective protein. It has the effect of protecting the spinal nerve of the body, avoiding the further injury of the spinal cord, and has a certain effect on the recovery of the spinal function of the later body.

【学位授予单位】：皖南医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R651.2

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