中西药与基因治疗家兔退变颈椎间盘对软骨终板钙化影响的研究

Study on the Effect of Combined Therapy of Chinese and Western Medicines and Gene Therapy on Cartilage End-plate Calcification in Rabbits with Cervical Intervertebral Disc Regression

目的了解家兔颈椎软骨终板钙化与颈椎间盘退变的关系,探讨抗骨增生胶囊与葡立胶囊联合应用及基因治疗退变颈椎间盘对软骨终板钙化有无抑制作用。方法选用35只4月龄新西兰兔,建立家兔颈椎动力平衡失调模型,诱导颈椎间盘退变(正常对照组不作处理)。术后7个月,药物治疗组(浅层、全层)给予抗骨增生胶囊和葡立胶囊(剂量按体重折算),灌胃2次/日,连用1个月,基因治疗组则将带有转化生长因子β1(TGF-β1)基因的重组质粒DNA注入C2-3、C3-4、C4-5椎间盘中(每个椎间盘用量为20μl)。1个月后用耳缘静脉气栓法处死各组动物,切取C4-5椎间盘(包括上、下部分椎体),在形态学上评定颈椎间盘退变程度,测定各组动物颈椎软骨终板钙化层厚度,进行组间比较。结果模型浅层组、模型全层组与正常对照组比较,颈椎软骨终板钙化层明显增厚,有统计学差异,而抗骨增生胶囊与葡立胶囊联合应用、基因治疗退变颈椎间盘对软骨终板钙化有明显抑制作用(P<0·05)。结论软骨终板钙化是颈椎间盘退变的始动因素,二者呈高度正相关。抗骨增生胶囊与葡立胶囊联合应用及基因治疗退变颈椎间盘对软骨终板钙化有明显的抑制作用。

Objective To study the relationship between cartilage end-plate calcification （CEC） and cervical intervertebral discs regression （CIDR） in rabbits, and to study the inhibitory effect of combined therapy of Kanggu Zengsheng Capsule （KZC） and Puli Capsule （PLC） and gene therapy （GT） on CEC by measuring the thickness of CEC layer. Methods Thirty-five New Zeland rabbits of 4 months old were selected to establish cervical dynamic imbalance rabbit model for inducing CIDR （No disposal was given to rabbits in the normal control group）. Seven months after operation, combined therapy of KZC and PLC were given, in doses calculated by body weight, to the modeled rabbits in the drug treated group with CEC of either superficial layer or full layer, twice a day by gavage for 30 successive days. While to those in the gene therapy group, the recombinant plasmid DNA with transforming growth factor-β1 （TGF-β1） was injected once to their intervertebral discs （ID） Of C2-3 ,C3-4 and C4-5, 20μl for each injection. One month later, all rabbits were sacrificed with periotic venous gas embolic method and their ID of C4-5（including partial body of the upper and lower vertebrae） was resected. The degree of CIDR was evaluated morphologically, and the thickness of CEC in rabbits was measured and compared between groups. Results Thickness of CEC in the model group, either of superficial layer or of full lay- er, was significantly more than that in the normal control group with significant difference. Both combined KZC and PLC therapy and gene therapy showed significant inhibitory effects on CEC in treating CIDR （P〈 0.05）. Conclusion CEC is the initial factor of CIDR with highly positive correlation. Both combined therapy of KZC and PLC and gene therapy can significantly inhibit CEC.