大鼠创伤弧菌脓毒症时外周血脂多糖受体mCD14的表达及其动态变化

The expression and the dynamic change of lipopolysaccharide receptor mCD14 in peripheral blood of the rats with Vibrio vulnificus sepsis

目的 探讨大鼠创伤弧菌(Vibrio vulnificus,VV)脓毒症外周血中膜性脂多糖(LPS)受体mCD14的表达及其动态变化.方法 Sprague-Dawley (SD) 雄性大鼠下肢皮下注射VV,造成VV脓毒症模型,用兔抗鼠CD14抗体和异硫氢酸荧光素(FTTC)标记的羊抗兔抗体对外周血细胞进行免疫染色,流式细胞仪测定FITC-CD14阳性细胞数及其荧光强度,同时观察染菌大鼠行为学改变情况.结果 染菌后4h,大鼠开始出现活动减少,呼吸急促,皮温升高,下肢肿胀,后症状进行性加重,12 h后染毒大鼠出现紫绀、间断抽搐等.注射VV后6 h,9 h、12 h、16 h mCD14表达均升高,FITC-CD14细胞阳性数达(39.849±3.792)%、(39.0474±3.146)%、(39.360±5.280)%、(44.864 4±4.374)%(均P＜0.01),荧光强度增强.FITC-CD14细胞阳性数在0～6h升高迅速,6-12 h保持在较高水平的平台期,而12～16 h及其以后其又继续升高.结论 创伤弧菌脓毒症血中mCD14的表达水平随脓毒症的病情进展而升高,其与脓毒症的发生、发展密切相关,动态监测其改变对创伤弧菌脓毒症发病过程具有一定的预警意义.

Objective To detect the expression and the dynamic change of lipopolysaccharide(LPS)receptor mCD14 in the peripheral blood of the rats with Vibrio vulnificus sepsis. Methods Sixty male Sprague-Dawley (SD) rats were injected vibrio vulnificus ,which made vibrio vulnificus sepsis model,the cells in peripheral blood were stained by immunofluorescent method (CD14 rabbit polyclonal antibody and Fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit immunoglobulin specific polyclonal antibody) and analyzed with flow cytometry (FCM). The percentage and mean fluorescence intensity (FI) of CD14-positive cells were taken as the indexes. Meanwhile, the change in ethology was observed . Results The rats showed breathlessness, fervescence, and swelling in extremitas inferior at 4 h after being injected VV. These symptoms were worse and worse , and the rats displayed cyanosis and convulsion after 12 h.The expression of mCD14 increased significantly at 6 h, 9 h, 12 h, 16 h after being injected VV, and the FITC-CD14 positive cells were (39.849±3.792)%, (39.047±3.146)%, (39.360±5.280)%, (44.864±4.374)%(P<0.01),and their FI also strengthened. The increasing rate of The FITC-CD14 positive cells was fast before 6 h after being injected VV. The FITC-CD14 positive cells did not significantly change from 6 h to 12 h. However, after 12 h it heightened sequentially. Conclusion Our data suggest that mCD14 elevation is associated with the development of vibrio vulnificus sepsis. Monitoring the change of mCD14 in peripheral blood have the benefit of mastering the VV sepsis progress.