构建新西兰兔脊柱结核模型的实验研究

发布时间：2019-01-19 12:16

【摘要】：目的在新西兰兔腰5椎体下终板种植结核分枝杆菌H37Rv标准菌株,构建兔脊柱结核模型。方法65只新西兰兔随机分为3组,实验组30只,对照组30只,空白对照组5只。弗氏完全佐剂致敏1月后,在腰5椎体下终板钻孔,实验组注入1×107CFU·m L~(-1)结核杆菌混悬液0.1m L,对照组注入等量生理盐水,空白对照组不作任何处理。术后观察兔一般情况,术后1个月、2个月对造模兔行X线、CT检查,术后2个月处死动物,行大体解剖观察,病椎取材作石蜡切片,对CT为阳性表现的兔椎旁肉芽组织进行结核杆菌培养。结果实验组25只、对照组28只,空白对照组5只兔完成实验。X线检查:术后1个月,实验组36.0%(9/25)的兔腰5、6椎体出现轻度骨质破坏;术后2个月,实验组52.0%(13/25)的兔腰5、6椎体骨质破坏较前加重,1只出现脊柱后凸畸形;对照组未见椎体骨质破坏,椎间隙无变化。CT扫描:术后1个月,实验组48.0%(12/25)的兔腰5、6椎体骨质破坏,病椎内细小死骨形成;术后2个月,实验组72.0%(18/25)的兔腰5、6椎体骨质破坏较前加重,5只椎旁脓肿形成;对照组未见椎体骨质破坏,无椎旁脓肿形成。术后2个月,实验组18只病椎石蜡切片苏木素-伊红(HE)染色显示有类上皮样细胞、结核坏死灶形成。结合影像学与组织病理学,术后2个月,兔脊柱结核造模成功率为72.0%(18/25)。18只兔的椎旁肉芽组织(CT阳性表现)结核杆菌培养阳性率为38.9%(7/18)。结论弗氏完全佐剂致敏的新西兰兔腰5椎体局部种植H37Rv标准菌株,可成功构建兔脊柱结核模型。
[Abstract]:Objective to establish a spinal tuberculosis model in New Zealand rabbits by planting Mycobacterium tuberculosis H37Rv standard strain in the lower lumbar endplate of 5 vertebrae. Methods Sixty-five New Zealand rabbits were randomly divided into 3 groups: experimental group (n = 30), control group (n = 30) and blank control group (n = 5). One month after sensitization with Freund's complete adjuvant, 1 脳 107CFU ml ~ (-1) Mycobacterium tuberculosis suspension was injected into the lumbar 5 vertebrae lower endplate in the experimental group, the control group was injected with the same amount of normal saline, and the blank control group was not treated. After 1 month and 2 months after operation, the model rabbits were examined by X-ray and CT. The animals were killed 2 months after operation, the animals were observed by general anatomy, and the diseased vertebrae were taken as paraffin sections. Mycobacterium tuberculosis was cultured in paravertebral granulation tissue of rabbits with positive CT. Results 25 rabbits in the experimental group, 28 rabbits in the control group and 5 rabbits in the blank control group completed the experiment. X-ray examination showed that 36.0% (9 / 25) of the rabbits in the experimental group had slight bone destruction in the lumbar vertebrae of the experimental group 1 month after operation. 2 months after operation, 52.0% (13 / 25) of the rabbits in the experimental group suffered more serious bony destruction in the lumbar vertebrae, and 1 had kyphosis deformity. CT scan: 1 month after operation, 48.0% (12 / 25) of the rabbits' lumbar vertebrae in the experimental group suffered from bone destruction, and small dead bones were formed in the diseased vertebrae. Two months after operation, 72.0% (18 / 25) of the rabbits in the experimental group had more serious bone destruction and 5 paravertebral abscesses in the lumbar spine of the experimental group than before, while in the control group, there was no bony destruction and no paravertebral abscess. At 2 months after operation, hematoxylin-eosin (HE) staining in paraffin sections of 18 diseased vertebrae in the experimental group showed epithelioid cells and necrotic tuberculous foci. Combined with imaging and histopathology, the success rate of spinal tuberculosis modeling was 72.0% (18 / 25) at 2 months after operation, and the positive rate of Mycobacterium tuberculosis culture in paravertebral granulation (CT) was 38.9% (7 / 18) in 18 rabbits. Conclusion the H37Rv standard strain in the lumbar 5 vertebrae of New Zealand rabbits sensitized with Freund's complete adjuvant can successfully construct the spinal tuberculosis model in rabbits.
【作者单位】：宁夏医科大学总医院骨科;宁夏医科大学;宁夏医科大学总医院医学检验中心;
【基金】：国家自然科学基金(30960391)
【分类号】：R529.2;R-332

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