本文選題：骨成型蛋白-7(Bone-morphogenetic + protein　； 參考：《山東中醫(yī)藥大學(xué)》2017年碩士論文

【摘要】：目的:觀察針刺方法不同對(duì)兔椎間盤髓核變性程度的影響,從而建立一個(gè)更貼近臨床的動(dòng)物模型;分別通過影像學(xué)、解剖學(xué)和酶聯(lián)免疫吸附劑測定(enzyme linked immunosorbent assay,ELISA),驗(yàn)證體內(nèi)注射骨成型蛋白-7(Bone-morphogenetic protein-7,BMP-7)對(duì)兔退變椎間盤髓核內(nèi)Ⅱ型膠原蛋白含量的影響,為進(jìn)一步研究多因素對(duì)退變椎間盤治療效果提供參考,為臨床治療腰椎退行性變提供新思路、新方法。方法:準(zhǔn)備18只成年健康新西蘭大白兔,每只兔均選取L2~L6之間的全部4個(gè)椎間盤為研究對(duì)象,將18只兔隨機(jī)地分成3組,A組6只,對(duì)其椎間盤既穿刺且旋轉(zhuǎn);B組6只,對(duì)其椎間盤僅穿刺,不旋轉(zhuǎn);C組6只為空白組,不做任何特殊處理,分別于術(shù)后1周、2周、4周從每組中隨機(jī)抽取2只,驗(yàn)證兔椎間盤髓核退行性變模型。模型驗(yàn)證成功后,取成年健康新西蘭大白兔48只,每只兔均選取L2~L6之間的全部4個(gè)椎間盤為研究對(duì)象,將48只兔隨機(jī)分成4組并給予編號(hào)進(jìn)行造模。其中治療組12只,在退變椎間盤髓核內(nèi)及周圍注射BMP-7,術(shù)后三天予肌注抗生素預(yù)防感染,不做其他處理;設(shè)置對(duì)照組12只,在退變椎間盤髓核內(nèi)及周圍注射等滲生理鹽水,術(shù)后三天予肌注抗生素預(yù)防感染,不做其他處理;空白組12只,僅完成造模,對(duì)應(yīng)時(shí)間同樣給予肌注抗生素,以便排除抗生素對(duì)治療結(jié)果的干擾;另外,設(shè)置一組鎮(zhèn)靜組與治療組對(duì)照研究,在其完成治療組相同治療方案的同時(shí),給予持續(xù)鎮(zhèn)靜處理,以減少兔的運(yùn)動(dòng)。分別于造模當(dāng)天、造模后2周(既造模成功當(dāng)天)、治療后2周(既造模后4周)、治療后6周(既造模后8周)進(jìn)行檢驗(yàn)對(duì)比,空白組亦于相應(yīng)時(shí)間進(jìn)行檢驗(yàn)對(duì)比,這樣,每組又縱向分為4個(gè)亞組,每個(gè)亞組各3只兔的12個(gè)椎間盤作為研究對(duì)象,對(duì)相應(yīng)亞組的檢驗(yàn)結(jié)果進(jìn)行橫向和縱向比較,檢驗(yàn)方法有(1)體重變化情況,(2)X線攝影記錄椎間盤高度,(3)解剖學(xué)觀察,(4)ELISA檢測椎間盤髓核內(nèi)Ⅱ型膠原蛋白含量。實(shí)驗(yàn)結(jié)果:通過對(duì)觀察指標(biāo)和統(tǒng)計(jì)學(xué)結(jié)果的總結(jié),A組模型的退變程度與B組相近,但造模成功率高,可重復(fù)性穩(wěn)定可靠,造模操作2周后即可形成理想退變模型。治療情況,在橫向比較中:X線片表現(xiàn)方面,治療組在椎間盤高度上大于對(duì)照組和空白組,具有統(tǒng)計(jì)學(xué)意義,對(duì)照組和空白組之間未見明顯差異;解剖學(xué)觀察方面,治療組椎間盤較新鮮,結(jié)構(gòu)較清楚,瘢痕填塞量少,而對(duì)照組和空白組椎間髓核被瘢痕組織填塞較明顯,對(duì)照組和空白組之間未見明顯差異;ELISA檢測Ⅱ型膠原蛋白含量方面,治療組高于對(duì)照組和空白組,具有統(tǒng)計(jì)學(xué)意義,對(duì)照組和空白組之間未見明顯差異。在縱向比較中:治療組椎間盤X線片上高度變窄程度小,解剖學(xué)觀察瘢痕化進(jìn)度不明顯,ELISA檢測Ⅱ型膠原蛋白含量丟失不明顯,無統(tǒng)計(jì)學(xué)意義;對(duì)照組及空白組X線片高度變窄程度明顯,解剖學(xué)檢查可見髓核內(nèi)瘢痕組織逐漸增多,ELISA檢測Ⅱ型膠原蛋白含量流失明顯,具有統(tǒng)計(jì)學(xué)意義。鎮(zhèn)靜組與治療組各項(xiàng)結(jié)果均無明顯區(qū)別,統(tǒng)計(jì)分析結(jié)果顯示無統(tǒng)計(jì)學(xué)意義。結(jié)論:對(duì)兔椎間盤既穿刺且旋轉(zhuǎn),可以制造良好的椎間盤髓核退行性變模型,造模操作2周后即可形成理想退變模型;BMP-7能有效緩解兔退變椎間盤髓核內(nèi)Ⅱ型膠原蛋白含量的丟失,體內(nèi)治療兔椎間盤髓核退行性變具有確定效果,鎮(zhèn)靜輔助治療對(duì)退變的進(jìn)程沒有明顯幫助。
[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.
【學(xué)位授予單位】：山東中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R681.5

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