發(fā)布時(shí)間：2018-06-12 19:11

  本文選題：椎間盤(pán)退變 + 基因治療　； 參考：《重慶醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的 研究兔腰椎間盤(pán)髓核內(nèi)注射慢病毒攜帶的BMP-2和細(xì)胞分化抑制因子1(inhibitor of differentiation 1,Id1)基因能否有效改善椎間盤(pán)退變進(jìn)程。方法 (1)經(jīng)腹建立兔腰椎間盤(pán)細(xì)針穿刺退變模型,并在影像學(xué)和組織學(xué)上加以驗(yàn)證;(2)取3月齡新西蘭兔32只,體質(zhì)量2.0~2.5 kg;以L3/4、L4/5、L5/6為目標(biāo)椎間盤(pán),經(jīng)腹細(xì)針穿刺法制造椎間盤(pán)退變模型。將30只造模成功的實(shí)驗(yàn)動(dòng)物隨機(jī)分為5組,每組6只;造模后4周于各組目標(biāo)椎間盤(pán)中分別注射Lv-BMP-2病毒(A組)、Lv-BMP-2+Lv-Id1病毒(B組)、Lv-Id1病毒(C組)、Lv-綠色熒光蛋白空載體病毒(D組)及PBS(E組)。于注射后2、4、8周各組分別隨機(jī)取2只動(dòng)物行T2-mapping MRI檢查獲得確切的T2弛豫時(shí)間(T2值);之后處死動(dòng)物取椎間盤(pán)組織,應(yīng)用實(shí)時(shí)熒光定量PCR、ELISA法檢測(cè)Ⅱ型膠原及蛋白聚糖的m RNA相對(duì)表達(dá)量及含量。結(jié)果 (1)手術(shù)操作本身對(duì)兔子的體質(zhì)量無(wú)明顯影響,僅一只兔子發(fā)現(xiàn)腹壁與腸管輕度部分粘連;術(shù)后證實(shí)出穿刺的椎間盤(pán)為目標(biāo)椎間盤(pán),其椎間盤(pán)高度指數(shù)和MRI信號(hào)強(qiáng)度呈進(jìn)行性退變改變;穿刺后4周組織學(xué)表現(xiàn)為髓核區(qū)域脊索細(xì)胞顯著減少而纖維細(xì)胞增生,纖維環(huán)結(jié)構(gòu)紊亂。(2)T2-mapping MRI檢查示,注射后0、2周各組間T2值比較差異均無(wú)統(tǒng)計(jì)學(xué)意義(P0.05);注射后4周,A、B組T2值顯著高于C、D、E組(P0.05),A、B組間比較差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05);注射后8周,B組T2值顯著高于其余各組(P0.05)。實(shí)時(shí)熒光定量PCR及ELISA檢測(cè)示,注射后2、4、8周B組Ⅱ型膠原及蛋白聚糖的m RNA相對(duì)表達(dá)量及含量均顯著高于其余各組(P0.05)。結(jié)論 (1)經(jīng)腹纖維環(huán)細(xì)針穿刺法,操作簡(jiǎn)單,目標(biāo)椎間盤(pán)定位準(zhǔn)確,手術(shù)視野更佳,椎間盤(pán)退變程度一致,并發(fā)癥少;而且在影像學(xué)和組織學(xué)上符合人腰椎間盤(pán)退變情況;(2)聯(lián)合應(yīng)用BMP-2和Id1基因在體內(nèi)實(shí)驗(yàn)中能有效延緩?fù)醚甸g盤(pán)退變的進(jìn)程。
[Abstract]:Objective to investigate whether injecting lentivirus-carrying BMP-2 and 1(inhibitor of differentiation 1 / Id1 gene into the lumbar disc nucleus of rabbits can effectively improve the process of disc degeneration. Methods 1) A fine needle puncture model of lumbar intervertebral disc degeneration was established through abdomen and verified by imaging and histology. 32 New Zealand rabbits of 3 months old with a body mass of 2.0 ~ 2.5 kg were taken as the target disc, and L3 / 4 / 4 / 5 / 5 / 6 of the intervertebral disc was used as the target. The model of intervertebral disc degeneration was established by transabdominal fine needle puncture. Thirty successful experimental animals were randomly divided into 5 groups (6 in each group), and Lv-BMP-2 Lv-ID1 virus group B (Lv-BMP-2 Lv-ID1 virus group B) and PBSE group (group D) were injected with Lv-BMP-2 virus in each group 4 weeks after the model was established, and Lv-BMP-2 Lv-ID1 virus group B was injected into the target disc of each group 4 weeks after the model was established, and Lv-BMP-2 Lv-ID1 virus group B was treated with Lv-BMP-2 Lv-ID1 virus group B, respectively. At 8 weeks after injection, 2 animals in each group were randomly selected for T 2-mapping MRI to obtain the exact T 2 relaxation time and T 2 value. The relative expression and content of mRNA in collagen 鈪，

本文編號(hào)：2010768