益气化瘀方加葡糖胺和基因治疗对家兔退变颈椎间盘内白细胞介素1β和肿瘤坏死因子α含量的影响(英文)

Effect of anti-hyperosteogeny capsule combined with glucosamine hydrochloride capsule and gene therapy on the contents of interleukin-1 beta and tumor necrosis factor alpha in degenerated cervical intervertebral discs of rabbits

背景:退变颈椎间盘组织可释放多种细胞因子和炎症介质,它们在颈椎病的发生、发展过程中起重要作用。目的:观察抗细胞因子的中西药(益气化瘀方抗骨增生胶囊和葡糖胺)联合应用、基因治疗对家兔退变颈椎间盘内细胞因子含量的影响。设计:随机对照观察。单位:河北省人民医院骨科,河北省人民医院中心实验室。材料:实验于2003-03/2004-01在河北省人民医院中心实验室完成。选用新西兰兔35只,雌雄不拘,按随机数字表法分为7组,即正常对照组、模型浅层组、模型全层组、药物治疗浅层组、药物治疗全层组、基因治疗浅层组及基因治疗全层组,每组5只。方法:除正常对照组外,其余6组家兔均通过切除颈背部肌肉,建立颈椎动力平衡失调模型,诱导颈椎间盘退变。浅层组切除颈背浅肌群,全层组切除颈背浅肌群和深肌群。术后7个月,①药物治疗组(浅层、全层)家兔给予抗骨增生胶囊(由狗脊、骨碎补、鸡血藤、莱菔子、牛膝、女贞子、肉苁蓉、熟地黄、淫羊藿等成分组成,江苏康缘药业股份有限公司提供)和葡糖胺(山西中远威药业有限公司提供),分别溶于20mL蒸馏水中,调配成糊状后分别按1.1g/kg和0.06g/kg灌胃,2次/d,连用1个月。②基因治疗组(浅层、全层)家兔麻醉后,将带有转化生长因子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)。造模后8个月处死各组动物,切取C3～4,C4～5椎间盘作标本,用双抗夹心ELISA法检测各组动物颈椎间盘中白细胞介素1β、肿瘤坏死因子α含量。主要观察指标:造模后8个月各组家兔颈椎间盘内白细胞介素1β、肿瘤坏死因子α含量。结果:实验过程中35只家兔无脱失,全部进入结果分析。造模后8个月各组家兔颈椎间盘内白细胞介素1β、肿瘤坏死因子α含量比较:模型浅层组和模型全层组家兔显著高于正常对照组(P<0.05～0.01),药物治疗浅层组、基因治疗浅层组显著低于模型浅层组(P<0.05),药物治疗全层组、基因治疗全层组显著低于模型全层组(P<0.05～0.01)。结论:退变颈椎间盘组织释放多种细胞因子和炎症介质,它们可加速颈椎间盘退变。中西药联合应用及基因治疗能对其含量起明显的抑制作用。

BACKGROUND： Degenerated cervical intervertebral discs can release many kinds of cytokines and inflammatory mediums, which plays an important role in onset and development of cervical syndrome. OBJECTIVE： To observe the effect of anti-cytokine Chinese herb combined with western drug （anti-hyperosteogeny capsule combined with glucosamine hydrochloride capsule） and gene therapy, on the contents of interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α） of degenerated cervical intervertebral discs in rabbits. DESIGN： Randomized controlled observation. SETTING： Department of Orthopaedics and Central Laboratory of Hebei People＇s Hospital. MATERIALS： The experiment was carried out in the Central Laboratory of Hebei People＇s Hospital from March 2003 to January 2004. A total of 35 New Zealand rabbits of both genders were selected and divided into 7 groups according to randomly digital table： normal control group, shallow-layer model group, whole-layer model group, shallow-layer medicine group, whole-layer medicine group, shallow-layer gene group and whole-layer gene group. There were 5 rabbits in each group. METHODS： Except normal control group, rabbits in other 6 groups were used to establish models of dynamic dysequilibrium of cervical vertebra through cutting muscles of cervical back to induce degeneration of cervical intervertebral discs. Superficial group of muscle in cervical region was resocted in shallow-layer groups, and both superficial and deep group of muscle was resected in whole-layer groups. （1） Seven months after operation, rabbits in medicine groups were respectively treated with 1.1 g/kg anti-hyperosteogeny capsule （consisting of gouji, gusuibu, jixueteng, laifuzi, niuzi, nvzhenzi, roucongrong, shudihuang and yinyanghuo, etc.; Jiangsu Kangyuan Pharmaceutical Company Limited） and 0.06 g/kg glucosamine （Shanxi Zhongyuanwei Pharmaceutical Company Limited）, which was dissolved into 20 mL distilled water. The medicine was administrated twice a day for 1 month. （2） After anesthetizafion of rabbits in gene groups, recombinant plasmid DNA combined with transforming growth factor-β1 （TGF- β1） was injected to C2-3, C3-4 and C4-5 intervertebral disc （20μL per intervetebral disk）. Eight months after modeling, rabbits in each group were sacrificed and C3-4 and C4-5 intervertebral disc was used as samples. In addition, contents of IL-1β and TNF-α were measured with double-antibodies-sandwich-ELISA method. MAIN OUTCOME MEASURES： Contents of IL-1β and TNF-α in cervical intervertebral discs of rabbits at 8 months after modeling. RESULTS： All 35 rabbits were involved in the final analysis. Eight months after modeling, for contents of IL-1β and TNF-α, shallow-layer model group was higher than normal control group （P 〈 0.05-0.01）; shallow-layer medicine group and shallow-layer gene group were obviously low er than shallow-layer model group （P 〈 0.05）; whole-layer medicine group and whole-layer gene group were lower than whole-layer model group （P 〈 0.05-0.01）. CONCLUSION： The degenerated intervertebral discs can release many kinds of cytokine and inflammatory mediators which can enhance the degeneration of cervical intervertebral discs. Antihyperosteogeny capsule combined with glucosamine hydrochloride capsule and gene therapy can obviously reduce the contents of IL-1β and TNF-α in degenerated cervical intervertebral discs.