直立體位下無創(chuàng)軸向加載建立兔椎間盤退變動(dòng)物模型
發(fā)布時(shí)間:2018-06-25 22:04
本文選題:椎間盤退變 + 動(dòng)物模型。 參考:《中國脊柱脊髓雜志》2017年06期
【摘要】:目的:通過直立體位下無創(chuàng)性軸向加載的方式,建立一種新型兔腰椎間盤早期退變的動(dòng)物模型。方法:24只4月齡雄性新西蘭大白兔隨機(jī)分為實(shí)驗(yàn)組及對(duì)照組。將實(shí)驗(yàn)組動(dòng)物置于特制筒內(nèi)使之保持直立體位,并自頸部施以600g軸向載荷,每日6h;對(duì)照組動(dòng)物不接受任何處理而常規(guī)飼養(yǎng)。在實(shí)驗(yàn)開始前及實(shí)驗(yàn)開始后4周、8周、12周時(shí)對(duì)全部動(dòng)物行X線及MRI檢查,觀察實(shí)驗(yàn)組動(dòng)物在筒內(nèi)時(shí)腰椎的骨性結(jié)構(gòu),通過椎間盤高度指數(shù)(disc height index,DHI)測量,比較兩組動(dòng)物側(cè)臥位時(shí)L2/3、L4/5和L6/7節(jié)段椎間隙高度改變;通過髓核相對(duì)灰度值測量比較兩組動(dòng)物髓核含水量變化。14周后全部動(dòng)物處死,取L5/6節(jié)段膠凍狀髓核組織,應(yīng)用rtPCR檢測Ⅰ型膠原、Ⅱ型膠原及蛋白多糖表達(dá)水平;取L6/7節(jié)段椎間盤連同上、下各1mm軟骨下骨,應(yīng)用HE及天狼星紅染色觀察各組動(dòng)物椎間盤組織結(jié)構(gòu)改變。結(jié)果:2只實(shí)驗(yàn)組動(dòng)物在實(shí)驗(yàn)過程中死亡,其對(duì)應(yīng)的實(shí)驗(yàn)數(shù)據(jù)從結(jié)果中剔除。腰椎X線片顯示,在直立體位負(fù)重條件下,實(shí)驗(yàn)組兔腰椎形成明顯后凸,較側(cè)臥位下腰椎間隙明顯變窄,在L2/3節(jié)段,直立體位下椎間隙高度為側(cè)臥位時(shí)的75.1%(P0.05);在L4/5節(jié)段為54.8%(P0.05);在L6/7節(jié)段為47.9%(P0.05)。DHI測量顯示兩組動(dòng)物腰椎各節(jié)段DHI在任何時(shí)間點(diǎn)均無顯著差異(P0.05)。MRI結(jié)果顯示,在L2/3節(jié)段,實(shí)驗(yàn)組與對(duì)照組髓核灰度值在任何時(shí)間點(diǎn)均無統(tǒng)計(jì)學(xué)差異(P0.05),在L4/5節(jié)段,實(shí)驗(yàn)組與對(duì)照組髓核相對(duì)灰度值在12周時(shí)開始具有顯著差異(P0.05);在L6/7節(jié)段,兩組間在8周時(shí)即開始有顯著性差異(P0.05)。rt-PCR結(jié)果顯示,實(shí)驗(yàn)組Ⅰ型膠原m RNA表達(dá)明顯高于對(duì)照組(3.57倍,P0.05);而Ⅱ型膠原及蛋白多糖表達(dá)明顯低于對(duì)照組(分別為0.35倍和0.43倍,P0.05)。病理學(xué)檢查顯示對(duì)照組與實(shí)驗(yàn)組椎間盤組織結(jié)構(gòu)存在顯著差異,對(duì)照組髓核組織與纖維環(huán)間邊界清晰,髓核內(nèi)富含均勻分布的髓核細(xì)胞;實(shí)驗(yàn)組內(nèi)層纖維環(huán)明顯增生,而髓核區(qū)域相應(yīng)減小。結(jié)論:直立體位下無創(chuàng)性軸向加載方式可顯著加速兔腰椎間盤的退變進(jìn)程,其在發(fā)病機(jī)理上與人類椎間盤退變更加相似,該腰椎間盤退變動(dòng)物模型操作簡單、可重復(fù)性強(qiáng)。
[Abstract]:Aim: to establish a new animal model of early degeneration of lumbar intervertebral disc in rabbits by noninvasive axial loading in upright position. Methods 24 4 month old male New Zealand white rabbits were randomly divided into experimental group and control group. The animals in the experimental group were placed in a special tube and kept upright, and 600g axial load was applied from the neck for 6 hours per day, while the control group was kept without any treatment. X-ray and MRI examinations were performed before the beginning of the experiment and 4 weeks and 8 weeks to 12 weeks after the experiment. The bone structure of the lumbar vertebrae in the experimental group was observed. The disc height index (disc height) was used to measure the bone structure of the lumbar vertebrae. The changes of intervertebral space height in L2 / 3, L4 / 5 and L6 / 7 segments were compared between the two groups, and the changes of water content in the nucleus pulposus of the two groups were compared by measuring the relative gray value of the nucleus pulposus. After 14 weeks, all the animals in the two groups were killed and the gelatinized nucleus pulposus tissues of L5 / 6 segments were taken. The expression levels of collagen type 鈪,
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