兔脊柱结核模型建立的实验研究

An experimental study on modeling of rabbit spinal tuberculosis

目的建立H37Rv结核分支杆菌诱导的兔脊柱结核模型。方法选择未致敏处理的新西兰大白兔48只,在第5腰椎近椎间盘处钻孔,并填充明胶海绵,注射0.5 mg/0.1 m L结核菌悬液,术后观察新西兰大白兔一般情况,并用影像学、组织病理学、细菌学等检查对兔脊柱结核模型进行评估。结果受H37Rv结核菌株感染后的新西兰大白兔,局部反应较明显,全身反应较轻。48只兔中38只兔完成实验,10只因死亡、术后截瘫被淘汰,15只兔分别于术后第5、6周出现进食欠佳、消瘦,但生命体征平稳,其中4只术区局部肿胀,6只出现下肢运动障碍;其余23只兔术后进食较好,体质量未见明显变化。术后3个月行外科手术,暴露致病兔椎体,38只兔中29只兔椎体可见虫噬样改变,9只兔椎体未见明显变化。术中取脓肿形成的兔脓液培养示结核分枝杆菌生长;取周围软组织、脓壁行苏木精-伊红色(HE)染色,示坏死灶形成、较多炎细胞聚集、淋巴细胞、较少上皮样细胞,骨结构紊乱或消失。建立模型成功率为76.3%(29/38)。结论使用0.5 mg/0.1 m L的H37Rv标准菌株感染的新西兰白兔结核模型建立成功。

Objective To build a spinal tuberculosis model by drilling and planting mycobacterium tuberculosis H37 Rv standard strains in the lumbar vertebra of New Zealand white rabbits to provide animal models for future fundamental researches on bone tuberculosis. Methods Drill bone holes on the top of the 5th lumbar vertebra near the intervertebral disk of 48 non-sensitized New Zealand white rabbits and fill in gelatin sponge to absorb TB suspension（ bacteria amount： 0. 5 mg /0. 1 m L） for modeling,and set the vacant group. After the operation,inspect and confirm the models by general observation,images,histopathology,bacteriology and other means. Results There were significant local reactions in H37 Rv tuberculosis-infected New Zealand rabbits,but with a slightly systematic response. 38 rabbits finished the trial（38 /48） and 10 rabbits were eliminated due to deaths and postoperative paraplegia. And 15 rabbits had the signs of poor diet,marasmus and stable vitality（15 /38） at week 5 and 6respectively after operation,4 of which（4 /15） had local swelling in the operation area and 6 of lower limb dyskinesia（6 /15）,and the rest 23 rabbits were stable and had good diet,with unchanged quality（23 /38）. The surgery was conducted 3 months after to explore the pathogenic vertebrae of rabbits,and insect-bite changes were found in 29 rabbits（29 /38）. There were not any significant changes in the vertebrae of 9 rabbits. The vertebrae were exposed 3 months after operation,29 of which were ill,with insect-bite changes in the exposed vertebrae at different levels. The pus culture from the abscess taken from the operation showed the growth of mycobacterium tuberculosis. The peripheral soft tissues and pus walls were taken for HE staining,which showed the formation of necrosis,more infected cells aggregation,less lymphocytes,and the epithelioid cells and bone structure were disordered and disappeared. The success rate of modeling is 76. 3%（29 /38）. Conclusion It proved successful to set up tuberculosis models for New Zealand white rabbits with 0. 5mg /0. 1m L of H37Rv-infected standard strains.