【摘要】 目的鼻腔滴注法建立小鼠肺炎链球菌性脑膜炎模型,为研究肺炎链球菌致脑膜炎的分子机制奠定基础。方法将冻存的TIGR4在新鲜TSA血平板上培养16 h～18 h后,刮取细菌并稀释成3种不同密度的菌液,每种密度分成2份,其中1份加入一定量的透明质酸酶,另1份不加。对小鼠行浅麻醉,用卡介苗注射器将上述菌悬液缓缓滴入小鼠鼻腔,待其自动吸入,每只小鼠50μl。每天观察小鼠情况,分别于感染后24、48和72 h处死小鼠,取心脏血和一半脑组织匀浆,做细菌计数,另一半脑组织用于组织病理学检查。结果加透明质酸酶的TIGR4 3个剂量组小鼠脑膜炎发生率分别为20.00%、65.00%和46.67%,对照组分别为13.33%、53.33%和66.67%,差异无统计学意义(P= 1.000 0);其中3×10~7CFU组和3×10~8CFU组差异无统计学意义(P=0.462 1),两组合并后与3×10~6CFU组差异有统计学意义(P=0.0196)。结论1)透明质酸酶不能促进鼻腔滴注TIGR4致小鼠脑膜炎的发生;2)小鼠脑膜炎的发生率与鼻腔感染TIGR4剂量有关,适宜剂量为3×10~7CFU,低于该剂量脑膜炎的发生率显著降低。更多还原

【Abstract】 Objective To establish a mice meningitis model infected with Streptococcus pneumoniae.Methods TIGR4 was inoculated from frozen stocks onto tryptic soy agar(TSA)plates supplemented with 5% defibrinated rabbit blood and incubated for 16 h～18 h at 37℃.Bacteria was scraped from fresh plates and suspended in phosphate-buffered saline(PBS).They were divided into two parts,and added with or without hyaluronidase,respectively.BALB/c mice were anesthetized and infected with 50μl of the suspension via intranasal instillation.At 24 h,48 h and 72 h post-infec- tion respectively,mice were sacrificed,blood and half of brains were recovered for bacterial enumeration,and the other half of brain for histopathologie study.Results The incidence of meningitis after inoculation with different bacteria number added with hyaluronidase were 20.00%,65.00% and 46.67%,respectively.Those of control group were 13.33%,53.33% and 66.67%,respectively.There was not significant difference between the two groups with and without hyaluronidase(P=1.000 0),and also no significant difference between the two groups with 3×10~7CFU and 3×10~8CFU(P=0.462 1).But the difference between these two groups and 3×10~6CFU was significant(P=0.019 6). Conclusion Hyaluronidase was not a necessary factor to establish a pneumococcal meningitis via intranasal instillation, and the incidence of meningitis was related to the amount of TIGR4 via intranasal instillation.更多还原