应用纤维结合素建立新型兔腰椎间盘退变模型  

A novel rabbit disc degeneration model induced by fibronectin fragment

作　　者：刘海飞[1] 张晗[2] 乔光曦[1] 陶昊[1] 陈峰[1] 扈延龄[1] 王德春[1] 胡有谷[1] 

机构地区：[1]青岛大学医学院附属医院骨科,266003 [2]青岛大学医学院附属医院内科,266003

出　　处：《中华外科杂志》2013年第4期362-366,共5页Chinese Journal of Surgery

基　　金：国家自然科学基金资助项目（30271318,30672132）

摘　　要：目的应用兔椎间盘内显微注射纤维结合素片段（Fn-f）的方法建立新型动物椎间盘退变模型。方法34只新西兰大白兔随机分为3组：实验组（16只），L1-2L2-3，L3-4，L4-5，椎间盘髓核内显微注射30kDa的Fn-f25μl；对照组（16只）相应髓核内注射磷酸盐缓冲液（PBS）25汕l；空白组（2只）不做处理。术后4、8、12周处死实验组及对照组各4只兔，对髓核组织切片行番红O染色和HE染色，采用^35S整合分析法检测髓核蛋白多糖合成。实验组和对照组中各取4只兔分别于术前及术后4、8、12和16周行腰椎数字化x射线摄影（DR）检查，测量计算椎间盘高度指数（DHI），术后16周处死，行番红O染色、HE染色和蛋白多糖合成测定。空白组2只兔处死后检测髓核蛋白多糖合成。结果实验组兔椎间盘髓核细胞数目进行性减少，内层纤维环结构紊乱、破坏；髓核区4周后出现进行性缩小，术后16周髓核区消失，与内层纤维环组织不能分辨。实验组兔椎间盘蛋白多糖合成率进行性下降（F=263．241，P=0．000）；术后各时间点，实验组兔椎间盘蛋白多糖合成率均较对照组显著降低（t=-27．010～-2．833，P〈0．05）。术后8周DR示造模椎间隙开始出现明显狭窄，进行性加重，12周时椎间盘及临近椎体前方出现骨赘，16周时骨赘增生明显。术后4周，实验组与对照组DHI百分比（DHI％）分别为96．5％±1．7％和97．4％±1．2％，差异无统计学意义（P〉0．05）；术后8、12和16周，实验组DHI％分别为85．6％±3．8％、77．2％±3．5％和65．5％±5．6％，均较对照组明显减低（t=-21．225～-10．795，P〈0．01）。结论显微注射Fn-f可使兔腰椎间盘发生缓慢的、进行性的、与人椎间盘自然退变过程相似的退变，具有较好的模拟性和重复性，为椎间盘退变分子水平的研究提供了一种新型的、实用的动物模型。Objective To establish a novel and useful rabbit model of lumbar disc degeneration using microinjection of fibronectin fragment （Fn-f）. Methods Thirty-two New Zealand white rabbits underwent injection of N-terminal 30 kDa Fn-f （ experimental group） or phosphate buffered saline （PBS） （ control group） into the central region of L1-2, L2-3 , L3-4 , L4-5 discs using a 32-gauge microsyringe. Two rabbits （blank group） with no treatments were sacrificed to examine the proteoglycan synthesis of neucleus pulposus （NP） using 3SS_sulfate incorporation assay. At the 4-, 8-, 12-, and 16-week time points, the discs were examined histologically, radiographically, and with proteoglycan synthesis. Results Histology demonstrated a progressive loss of the cell numbers in NP and architecture destruction in NP and anulus fibrosus （AF） in Fn-f-injected discs over the 16-week study period. The NP regions in Fn-f-injected discs shrinked distinctly after the 4-week time point, and were not discernible with the inner AF by the 16-week time point. Protoglycan synthesis in Fn-f-injected discs decreased progressively （F = 263. 241, P = 0. 000）. At each time point, the Fn-f-injected discs showed significantly decreased proteoglycan synthesis compared with controls （t = - 27. 010- - 2. 833,P 〈 0.05）. The DHI% of the Fn-f-injected discs at the 4-, 8-, 12-,and 16-week time points were 96. 5% ± 1.7%, 85.6% ± 3.8%, 77. 2% ± 3.5% and 65.5% -＋ 5.6%, respectively. Comparing with the DHI% of PBS-injected discs （97.4% ± 1.2% ） , the Fn-f-injected discs exihibited no significant differences in disc heights at the 4-week time point （ P 〉 0. 05 ）, but significant decreases in disc heights at the 8-, 12-, and 16-week time points （ t = - 21. 225- - 10. 795, P 〈 0. 01 ）. Apparent anterior osteophytes formed at the 12-week time point and enlarged remarkablely by the 16-week time point in the experimental spines. Conclusions Fn-f can induce a progressively degenerative process in rabbit discs which is

关 键 词：疾病模型 动物 腰椎 椎间盘 纤维结合素 

分 类 号：R-332[医药卫生]