纤维环穿刺结合TNF-α注射构建大鼠尾椎间盘退变模型的实验研究  

An Experimental Study of the Construction of Caudal Intervertebral Disc Degeneration Model in Rats by Anulus Fibrosus Puncture Combined with TNF-α Injection

作　　者：夏炳江[1,2] 张磊[2] 胡松峰[1] 王宁[1] 侯明生[1] XIA Bingjiang ZHANG Lei HU Sonffeng et al(Shaoxing Hospital of Traditional Chinese Medicine, Zhejiang, Shaoxing 312000, China)

机构地区：[1]浙江省绍兴市中医院,浙江绍兴312000 [2]浙江中医药大学,浙江杭州310053

出　　处：《中国中医急症》2017年第7期1167-1171,共5页Journal of Emergency in Traditional Chinese Medicine

基　　金：浙江省绍兴市科技计划项目(2015B70060);浙江省中医药科技计划项目(2016ZB132)

摘　　要：目的观察X线透视下纤维环穿刺结合肿瘤坏死因子-α(TNF-α)椎间盘内注射构建大鼠尾椎间盘退变模型的可行性。方法取3月龄清洁级SD大鼠96只,随机分为空白对照组(A组)、对照组(B组)和实验组(C组)各32只。C组大鼠在X线透视辅助下,利用0.6 mm穿刺针穿刺尾椎5~6椎间隙(Co5/6),穿刺成功后,以微量注射器向椎间盘内注入TNF-α(10 ng/μL)25μL。B组大鼠在椎间盘穿刺成功后注入25μL生理盐水,A组大鼠不做任何处理。采用50%机械性撤足阈值、椎间隙相对高度、椎间盘组织学改变及椎间盘聚蛋白聚糖和Ⅱ型胶原基因表达评价大鼠尾椎间盘退变情况。结果造模后,C组大鼠50%机械性撤足阈值明显降低,椎间隙相对高度逐渐下降,HE染色显示椎间盘出现明显退变表现,椎间盘蛋白聚糖及Ⅱ型胶原mRNA表达下降,表明模型构建成功,与B组大鼠相比,C组大鼠诱发的椎间盘退变过程出现更早,进展更快,程度更重。结论 X线透视下纤维环穿刺结合TNF-α椎间盘内注射是一种可行的建立大鼠尾椎间盘退变模型的方法,其所诱发的椎间盘退变过程出现更早,进展更快,程度更重。Objective： To study the feasibility of the construction of caudal intervertebral disc degeneration model in rats by anulus fibrosus puncture combined with TNF-α injection. Methods： 96 SD rats were divided randomly into the blank group（group A,n = 32）,the control group（group B,n = 32） and the experimental group（group C,n = 32）. In group C,the coccygeal intervertbral discs in rats at Co5-6 were punctured by 0.06 mm needle and received 25 μL of TNF-α（10 ng/μL）. In group B,the coccygeal intervertbral discs in rats at Co5-6 were punctured by 0.06 mm needle and received 25 μL normal saline. In group A,no interventions were taken. The caudal intervertebral disc degeneration was evaluated by 50% mechanical withdrawal threshold,intervertebral disc relative height,intervertebral disc histological changes and intervertebral disc proteoglycan and collagen type Ⅱgene expression. Results： After modeling,the 50% mechanical withdrawal threshold decreased;intervertebral disc relative height reduced;intervertebral disc histological changed,HE staining showed visible disc degeneration;the gene expression of proteoglycan and collagen type Ⅱ intervertebral disc decreased. All these showed the model was successful. Compared with group B,the process of intervertebral disc degeneration in group C appeared earlier,faster,and more serious. Conclusion： In X-ray fluoroscopy,anulus fibrosus puncture combined with TNF-αinjection is a feasible method for the establishment of caudal intervertebral disc degeneration model in rats. The process of intervertebral disc degeneration induced by this method appeared earlier,faster,and more serious.

关 键 词：椎间盘退变 TNF-Α 动物模型 针刺 

分 类 号：R285.5[医药卫生—中药学]