【摘要】 目的 :应用克隆测序技术结合观察急性病毒性心肌炎模型的发病过程及病理改变 ,从病原学及病理学角度鉴定柯萨奇病毒B3 亲心肌株 (CVB3m)诱导BALB/c小鼠急性病毒性心肌炎 (VMC)模型 ,为VMC模型提供科学的鉴定方法。方法 :以CVB3m腹腔注射诱导BALB/c小鼠心肌炎 ,分别于 3、5、7、10、14天取心肌 ,以RT -PCR法检测CVB3m,并通过质粒克隆测定cDNA序列 ;同时观察小鼠心肌病理组织学变化。结果 :感染小鼠心肌组织中RT -PCR扩增产物与已知CVB3mcDNA序列相应片段吻合度 >99 3% ;小鼠心肌病变从心肌细胞肿胀、少数淋巴细胞浸润到坏死和炎性病变随病程发展逐渐增多 ,第 10天达高峰 ,至第 14天炎症逐渐恢复 ,出现不同程度的纤维化。结论 :提示用克隆测序法鉴定为VMC发病机理及药物防治研究提供了稳定可靠的动物模型 ,并为研究CVB3m病毒感染小鼠后基因变异提供了参考依据。更多还原

【Abstract】 Objective: To determine the viral myocarditis model and to observe the development of the acute viral myocarditis.Methods: 30 BALB/c mice were inoculated intraperitoneally with coxsakiverius B\-\{3m\} (CVB\-\{3m\}).The mice were killed and their hearts were examined on day 3,5,7,10,14 post-inoculation.The CVB\-\{3m\} RNA were detected with reverse transcription-polmerase chain reaction (RT-PCR), the RT-PCR product were coloned in plasmid to test the DNA sequence.The myocardial histopathological damage were examined.Results: The DNA sequence was similar as CVB\-\{3m\} cDNA by 99.3 percent.The myocardcyte swelling,lymphocytes infiltrating and necrosis,inflammation were found increased with the development of myocarditis.The damages went up to the peak at day 10 after CVB\-\{3m\} infection.To day 14, the infllammation gradually decreased with the different degrees of fibrosis.Conclusions: The strain we used was right one,can induce the BALB/c mice myocarditis model.The method of colone and DNA sequence test was suggested to difine the strain for model.更多还原