HCMV感染介导的椎间盘组织退变模型的病理学观察

Human cytomegalovirus infection of Balb/c mice's lumber intervertebral disc tissues mediated models of disc degeneration and its observation of pathology

目的探讨HCMV感染Balb/c小鼠对椎间盘退变及病理学改变的模型研究,活体Balb/c小鼠模型导入组织学、基因表达、免疫学的研究,确定人巨细胞病毒注射到腹腔对椎间盘退变的影响及不同剂量的对照性研究。方法(1)随机选取6-8周SPF级Balb/c小鼠84只,建立五个不同感染量HCMVAD169株感染Balb/c小鼠模型雌雄各6只为感染组(A组),同时设立病毒灭活组(B组)和细胞对照组(C组)雌雄各6只;(2)间接ELISA法检测小鼠血清中的IgG、IgM抗体水平,取各组小鼠椎间盘组织进行病毒分离,采用PCR和RT-PCR分别检测HCMVUL83基因和UL54基因转录产物;(3)取各组小鼠椎间盘组织行病理学观察。结果A组和B组小鼠IgG抗体均为阳性,C组为阴性,小鼠IgM抗体仅A组中2×106PFU/ml和2×105PFU/ml感染组雌性小鼠为阳性;上述两感染组小鼠病毒分离、PCR及RT-PCR均为阳性,病理学可见椎间盘组织髓核细胞数目减少,排列不均匀,纤维环结构紊乱;雄性小鼠、其余A组、B组和C组均无明显变化。结论HCMV注射后似乎能逐渐诱导椎间盘退变,出现病理学改变;该模型对椎间盘退变的分子水平研究有显著的优势;进一步研究将会阐述HCMV介导椎间盘退变相关的分子学等机制和过程。

Objective To observe the change of pathology showing in the degeneration of HCMV infects Balb/c mice intervertebral disc （IVD） tissues, A histologic, gene expression and immunology study was conducted in vivo in a Balb/c mice model to determine the effect of injection of HCMV into the intervertebral disc along with various control doses. Methods （ 1 ） Pick up at random eighty - four six or eight - week - old Balb/c mice at the level of SPF ; establish models of 6 female and 6 male by 5 distinct doses of HCMV AD169 （group A）and establish inactive virus control group of 6 mice （group B）and cell control group of 6 mice（group C）. （2）Detect the level of IgG,IgM in mice＇s blood serum by means of indirect ELISA. Get IVD tissues from each group of mice to undertake viral isolation and assessment; detect HCMV＇s gene UL83 and UL54 respectively, and corresponding genes＇transcription by PCR and RT- PCR; （3）Take IVD tissues to pathological section with HE stain and then make observations. Results IgG in mice＇s blood serum of group A and B is masculine; group C is negative; IgM in miceg blood serum in virus groups of 2 × 106 and 2 × 105 PFU/ml is masculine; In the virus -infected groups above, virus is separated and PCR , RT- PCR are masculine in female mice, the pathological examination showed that the cells in nucleus pulposus reduced, arrangement unevened and annulus fibrosuses were in disorder, there were no special changes of pathology in Male mice, other dose group A, group B and C. Conclusion HCMV appears to induce a progressive degenerative process within the intervertebral disc after injection that resembles degenerative disc disease. This model has several significant advantages for the study of disc degeneration at the molecular level. Further studies are warranted to elucidate the mechanism and molecular events associated with HC- MV -mediated disc degeneration.