小鼠肺炎支原体肺炎模型的建立及组织病理学评分方法的应用

A mouse model of mycoplasma pneumoniae pneumonia

目的 试图建立一个由肺炎支原体 (MP)感染引起的小鼠的急性肺炎的动物模型和对肺炎程度进行量化评价的组织病理学评分方法。方法 BALB/C小鼠按体重随机分 8组 ,每组 12只 ,在 0、1、2d滴鼻感染MP菌液 ,在 0、1、2、3、4、6、8、14、2 1d分批宰杀 ,并分批设立滴注培养基的对照组 ,每组 4只 ,以 0～ 2 6分的组织病理学评分来确定肺部的炎症反应程度 ,评分包括支气管周围浸润部位的百分比、支气管周围浸润的数量、支气管管腔内的渗出、血管周围的浸润、实质性肺炎五方面 ,所有实验组动物的病原学诊断均做了肺匀浆MP培养和肺匀浆的MPPCR鉴定。结果 经接种菌液于感染后第 1天每 1例小鼠肺部均出现支气管肺炎的炎症反应 ,组织病理学评分平均为 6 .32分 ,于第 3、4天达到最重 ,为 11.6 0和 9.5 8分 ,以后逐渐减轻 ,于第 2 1天炎症基本消失。本实验组中动物的组织病理学评分为 0～ 19分。培养基对照组未出现肺部炎症反应。感染后 1、2、3、4d肺组织培养和PCR检测均有MP生长 ,检出率为 10 0 % (12 / 12 )。在 6、8、14、2 1d分别有 2 / 12、2 / 12、0 / 12、0 / 12检测出支原体。结论 经 3d滴鼻感染MP菌液 ,经组织病理学和肺组织培养和PCR检测证实可使小鼠出现明显的MP肺炎 ,小鼠急性MP肺炎的模型已经建立 ,?

Objective To establish a mouse model for the study of the acute phase of pulmonary infection by M.pneumoniae and to establish a histopathology scoring system which would dosely reflect the severity of pneumonia.Methods One hundred and thirty-two SPF BALB/C mice were randomly separated to eight experiment groups,every experiment group had twelve mouse.M.pneumoniae was introduced intranasally on day 0,1,and 2.Mice were sacrificed on day 0,1,2,3,4,6,8,14,21 and the control groupes were established respectively,every group had four mice,they were inoculated with sterile broth.The detection of M.pneumoniae by cultures and PCR of the lung tissue were performed.A histopathologic scoring system adapted from a hamster M.pneumonia model was applied to our model.Lung specimens were evaluated for the percentage of involved sites with peribronchiolar infiltrates,quality of infiltrates,luminal exudates,perivascular infiltrates,and parenchymal pneumonia,defined inflammatory changes in the lungs on a scale of 0-26(least to most severe).Results After inoculation of M.pneumoniae,pulmonary inflammation occurd on day 1 and the average score was 6.32.The inflammation was severiest on day 3,4 and the average score was 11.6 to 9.58 and the highest score was 19.Then it recovered gradually and on day 21 the inflammation was almost disappeared.All histopathologic scores ranged from 0 to 19.The control group had no inflammation of lung.The detection of MP by culture and PCR in the lung tissue on day1,2,3,4 were all positive,2/12,2/12,0/12,0/12 were positive on day 6,8,14,21.The results of PCR were same as results of the lung cultures.Conclusion After introduced intranasally with M.pneumoniae on day 0,1,and 2,the histopathologic scores and the detection of MP by culture and PCR in mice lung confirm that there were obvious pneumonia,histopathology scoring system can apparenty reflect the severity of pneumonia in relation to its doses.A mouse model of mycoplasma pneumoniae pneumonia is established.A histopathologic scoring system can define acute inflammatory changes in the lung in BLAB/C mice.