骨成型蛋白-7对兔腰椎间盘退行性变治疗效果的研究

发布时间：2018-06-01 14:00

本文选题：骨成型蛋白-7(Bone-morphogenetic + protein　；参考：《山东中医药大学》2017年硕士论文

【摘要】：目的:观察针刺方法不同对兔椎间盘髓核变性程度的影响,从而建立一个更贴近临床的动物模型;分别通过影像学、解剖学和酶联免疫吸附剂测定(enzyme linked immunosorbent assay,ELISA),验证体内注射骨成型蛋白-7(Bone-morphogenetic protein-7,BMP-7)对兔退变椎间盘髓核内Ⅱ型胶原蛋白含量的影响,为进一步研究多因素对退变椎间盘治疗效果提供参考,为临床治疗腰椎退行性变提供新思路、新方法。方法:准备18只成年健康新西兰大白兔,每只兔均选取L2~L6之间的全部4个椎间盘为研究对象,将18只兔随机地分成3组,A组6只,对其椎间盘既穿刺且旋转;B组6只,对其椎间盘仅穿刺,不旋转;C组6只为空白组,不做任何特殊处理,分别于术后1周、2周、4周从每组中随机抽取2只,验证兔椎间盘髓核退行性变模型。模型验证成功后,取成年健康新西兰大白兔48只,每只兔均选取L2~L6之间的全部4个椎间盘为研究对象,将48只兔随机分成4组并给予编号进行造模。其中治疗组12只,在退变椎间盘髓核内及周围注射BMP-7,术后三天予肌注抗生素预防感染,不做其他处理;设置对照组12只,在退变椎间盘髓核内及周围注射等渗生理盐水,术后三天予肌注抗生素预防感染,不做其他处理;空白组12只,仅完成造模,对应时间同样给予肌注抗生素,以便排除抗生素对治疗结果的干扰;另外,设置一组镇静组与治疗组对照研究,在其完成治疗组相同治疗方案的同时,给予持续镇静处理,以减少兔的运动。分别于造模当天、造模后2周(既造模成功当天)、治疗后2周(既造模后4周)、治疗后6周(既造模后8周)进行检验对比,空白组亦于相应时间进行检验对比,这样,每组又纵向分为4个亚组,每个亚组各3只兔的12个椎间盘作为研究对象,对相应亚组的检验结果进行横向和纵向比较,检验方法有(1)体重变化情况,(2)X线摄影记录椎间盘高度,(3)解剖学观察,(4)ELISA检测椎间盘髓核内Ⅱ型胶原蛋白含量。实验结果:通过对观察指标和统计学结果的总结,A组模型的退变程度与B组相近,但造模成功率高,可重复性稳定可靠,造模操作2周后即可形成理想退变模型。治疗情况,在横向比较中:X线片表现方面,治疗组在椎间盘高度上大于对照组和空白组,具有统计学意义,对照组和空白组之间未见明显差异;解剖学观察方面,治疗组椎间盘较新鲜,结构较清楚,瘢痕填塞量少,而对照组和空白组椎间髓核被瘢痕组织填塞较明显,对照组和空白组之间未见明显差异;ELISA检测Ⅱ型胶原蛋白含量方面,治疗组高于对照组和空白组,具有统计学意义,对照组和空白组之间未见明显差异。在纵向比较中:治疗组椎间盘X线片上高度变窄程度小,解剖学观察瘢痕化进度不明显,ELISA检测Ⅱ型胶原蛋白含量丢失不明显,无统计学意义;对照组及空白组X线片高度变窄程度明显,解剖学检查可见髓核内瘢痕组织逐渐增多,ELISA检测Ⅱ型胶原蛋白含量流失明显,具有统计学意义。镇静组与治疗组各项结果均无明显区别,统计分析结果显示无统计学意义。结论:对兔椎间盘既穿刺且旋转,可以制造良好的椎间盘髓核退行性变模型,造模操作2周后即可形成理想退变模型;BMP-7能有效缓解兔退变椎间盘髓核内Ⅱ型胶原蛋白含量的丢失,体内治疗兔椎间盘髓核退行性变具有确定效果,镇静辅助治疗对退变的进程没有明显帮助。
[Abstract]:Objective: To observe the effect of different acupuncture methods on the degree of intervertebral disc degeneration in rabbits, so as to establish a more close clinical animal model by imaging, anatomy and enzyme linked immunosorbent assay (enzyme linked immunosorbent assay, ELISA), to verify the body injection of bone morphogenetic protein -7 (Bone-morphogenetic protein-7, BMP-7). The effect of the type II collagen content in the nucleus pulposus of the rabbit degenerative intervertebral disc provides a reference for the further study of the effect of multiple factors on the treatment of degenerative intervertebral disc, and provides a new way of thinking for the clinical treatment of degenerative lumbar degeneration. Methods: 18 adult healthy New Zealand rabbits were prepared. All the 4 intervertebral discs were selected for each rabbit. 18 rabbits were randomly divided into 3 groups, group A 6, puncture and rotation of the intervertebral disc, group B, 6, only puncture and non rotation of the intervertebral disc, 6 group in group C, no special treatment. 2 rabbits were randomly selected from each group at 1 weeks, 2 weeks and 4 weeks respectively to verify the rabbit disc nucleus degeneration model. Model verification after success, 48 adult healthy New Zealand white rabbits were selected. All the 4 intervertebral discs between L2~L6 were selected for each rabbit, and 48 rabbits were randomly divided into 4 groups and given a model. 12 of the treatment groups were injected with BMP-7 in and around the nucleus and around the nucleus of degenerative intervertebral disc, and the antibiotics were injected to the muscle for three days after the operation. In the control group, 12 rats were injected with isotonic saline within and around the degenerative disc nucleus and injected with antibiotics to prevent infection and no other treatment on the three day after operation. 12 rats in the blank group only completed the model, and the antibiotics were given at the same time in order to exclude the interference of the antibiotics to the treatment results; in addition, a group of sedative groups and treatment groups were set up. According to the study, the same treatment regimen was completed in the treatment group, and the continuous sedation treatment was given to reduce the motion of the rabbit. 2 weeks after the model was built (the day of model success), 2 weeks after the treatment (4 weeks after the model), and 6 weeks after the treatment (8 weeks after the model), and the blank group was also tested and compared at the corresponding time. Each group was divided into 4 subgroups, and 12 intervertebral discs of 3 rabbits in each subgroup were used as the research subjects. The test results of the corresponding subgroups were compared horizontally and vertically. The test method had (1) body weight change, (2) X-ray photography recorded the height of intervertebral disc, (3) anatomical observation, (4) ELISA detection of type II collagen content in the nucleus of intervertebral disc. Experimental results: through the summary of the observation and statistical results, the degree of degeneration of the A group was similar to that of the B group, but the success rate of the model was high and the reproducibility was stable and reliable. The ideal degenerative model could be formed after 2 weeks of model operation. In the blank group, there was no significant difference between the control group and the blank group. In the anatomical observation, the intervertebral disc in the treatment group was fresh, the structure was clearer, the scar filling was less, but the intervertebral nucleus of the control group and the blank group was more obvious in the scar tissue, and there was no significant difference between the control group and the blank group; ELISA detection of type II collagen was found. The protein content, the treatment group was higher than the control group and the blank group, with statistical significance, there was no significant difference between the control group and the blank group. In the longitudinal comparison, the degree of height narrowing on the intervertebral disc in the treatment group was small, the anatomical observation of scar formation was not obvious, and the loss of type II collagen content was not obvious in ELISA examination, and no statistical meaning was statistically significant. The degree of height narrowing in the X-ray films of the control group and the blank group was obvious. The anatomical examination showed that the scar tissue in the nucleus of the medulla gradually increased. The ELISA detection type II collagen content loss was obvious. There was no significant difference between the results of the sedation group and the treatment group. Conclusion: the statistical analysis showed no statistical significance. Conclusion: the rabbit disc was not statistically significant. Conclusion: the rabbit intervertebral disc is not significant. A good degenerative model of intervertebral disc nucleus can be made by puncture and rotation, and an ideal degeneration model can be formed after 2 weeks of operation. BMP-7 can effectively alleviate the loss of type II collagen content in the nucleus pulposus of rabbit degenerative intervertebral disc. In vivo treatment of rabbit disc nucleus degeneration has a definite effect. There was no obvious help in the journey.
【学位授予单位】：山东中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R681.5

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