兔尿源性脓毒症过程中淋巴细胞功能变化的探索

To explore the functional changes of lymphocytes during the process of urine-derived sepsis in the rabbits

目的 观察兔尿源性脓毒症模型中不同时间点淋巴细胞功能变化。方法 2.0~2.5 kg新西兰雄兔18只,分为Control组(正常组)、Sham组(假手术组)和脓毒症1、3、5、7 d组,每组3只,通过结扎新西兰兔左侧输尿管远端并向输尿管内注入大肠杆菌悬液(ATC25922)建立兔尿源性脓毒症模型;Sham组仅给予暴露左侧输尿管中段,Control组不做任何处理。脓毒症组分别于造模后1、3、5、7 d取材,Control组和Sham组于第7天取材,检测各组炎症因子(TNF-α、PCT、IL-6、IL-1β、HMGB1)水平和免疫共刺激信号(CD28、CD40L、TIM-3、PD-1、CTLA4)mRNA水平;HE染色观察各组肾脏结构改变。结果 脓毒症炎症因子TNF-α、PCT、IL-6、IL-1β、HMGB1在第3天表达最高,免疫共刺激分子CD28、CD40L在第5天表达最高,免疫共抑制分子TIM-3、PD-1、CTLA4在第7天表达最高,肾脏病理损伤程度随时间延长不断加重。结论 兔尿源性脓毒症过程中,新西兰兔炎症反应于第3天最严重,第5天出现淋巴细胞活化增强,第7天即出现淋巴细胞功能耗竭。

Objective To explore the functional changes of lymphocytes at different time points in the rabbit models of urogenic sepsis.Methods 18 New Zealand male rabbits of 2.0-2.5 kg were divided into Control group,Sham group,group 1 d,group 3 d,group 5 d and group 7 d with three rabbits in each group.Rabbit model of urine-derived sepsis was established by ligating the distal end of the left ureter of New Zealand rabbits and injecting E.coli suspension(ATC25922)into the ureter.The left midureter was exposed in Sham group,but the Control group was not given any treatment.The samples in the sepsis group were taken at 1st,3rd,5th,7th day after modeling.The samples of the Control group and the Sham group were taken at the 7th day to detect the levels of inflammatory factors(HMGB1,IL-1β,IL-6,TNF-α,PCT)and the levels of immunocostimulatory signals(CD28,CD40L,TIM-3,PD-1,CTLA4)mRNA in each group.HE staining was used to observe the structural changes of the kidneys in each group.Results The expression of inflammatory factors(HMGB1,IL-1β,IL-6,TNF-α,PCT)were the highest in group 3 d,CD28 and CD40L were the highest in group 5 d,TIM-3,PD-1 and CTLA4 were the highest in group 7 d.The degree of renal pathological damage gradually worsened with the time.Conclusions In the process of urine-derived sepsis in the rabbits,the inflammatory response of New Zealand rabbits was most severe in group 3 d,with enhanced lymphocyte activation in group 5 d and lymphocyte depletion in group 7 d.