自然杀伤细胞及CD8^(+)T细胞缺失对泡球蚴感染小鼠肝功能的影响

Effect of absence of natural killer cells and CD8^(+)T cells on liver function in mice infected with metacestode of Echinococcus multilocularis

目的观察自然杀伤(NK)细胞、CD8^(+)T细胞的缺失对泡球蚴感染小鼠肝功能的影响。方法选取6~8周龄,体重为18~20 g的SPF级雌性C57BL/6小鼠共30只,按随机数字表法将其分为对照组、泡球蚴感染组、NK细胞清除组、NK细胞清除对照组、CD8^(+)T细胞清除组、CD8^(+)T细胞清除对照组,每组5只。泡球蚴感染组、NK细胞清除组、NK细胞清除对照组、CD8^(+)T细胞清除组、CD8^(+)T细胞清除对照组均经门静脉注射多房棘球绦虫头节(10个头节/μl,200μl),对照组经门静脉注射等量生理盐水。实验第4周,NK细胞清除组腹腔注射NK细胞清除抗体,100μg/次,1次/周,同时NK细胞清除对照组腹腔注射等量IgG2a同型对照抗体;CD8^(+)T细胞清除组腹腔注射CD8^(+)T细胞清除抗体200μg/次,1次/周,同时CD8^(+)T细胞清除对照组腹腔注射等量IgG2b同型对照抗体。实验第12周采集六组外周血分离血浆并检测小鼠肝肾功能指标总蛋白(TP)、碱性磷酸酶、谷丙转氨酶(GPT)、谷草转氨酶(GOT)、血清白蛋白(ALB)、乳酸脱氢酶、尿酸、尿素、肌酐。结果泡球蚴感染组外周血GPT、GOT高于对照组,差异有统计学意义(P<0.05或P<0.01)。NK细胞清除组外周血GPT、GOT高于NK细胞清除对照组,TP、ALB低于NK细胞清除对照组,差异有统计学意义(P<0.05或P<0.01)。CD8^(+)T细胞清除组外周血TP低于CD8^(+)T细胞清除对照组,差异有统计学意义(P<0.05)。结论泡球蚴感染会导致一定程度的肝功能损伤,尤其在NK细胞耗竭状态下肝功能损伤会进一步加重。

Objective To observe the effect of the absence of natural killer(NK)cells and CD8^(+)T cells on the liver function in mice infected with metacestode of Echinococcus multilocularis.Methods A total of 30 SPF grade female C57BL/6 mice aged six to eight weeks and with a body weight of 18-20 g were selected and divided into control group,metacestode of Echinococcus multilocularis infection group,NK cell clearance group,NK cell clearance control group,CD8^(+)T cell clearance group,and CD8^(+)T cell clearance control group according to random number table method,with five mice in each group.Metacestode of Echinococcus multilocularis infection group,NK cell clearance group,NK cell clearance control group,CD8^(+)T cell clearance group,CD8^(+)T cell clearance control group were all injected into the portal vein with the scolices of Echinococcus multilocularis Leuckart(10 scolices/μl,200μl),and the control group was injected into the portal vein with equal amount of normal saline.At the fourth week of the experiment,the NK cell clearance group was intraperitoneally injected with NK cell clearance antibody,100μg each time,once a week,while the NK cell clearance control group was intraperitoneally injected with the same amount of IgG2a homologous control antibody;CD8^(+)T cell clearance group was intraperitoneally injected with CD8^(+)T cell clearance antibody 200μg each time,once a week,while CD8^(+)T cell clearance control group was intraperitoneally injected with the same amount of IgG2b homologous control antibody.At the 12th week of the experiment,the peripheral blood of six groups was collected,the peripheral blood plasma was separated,and the liver and kidney function indexes of total protein(TP),alkaline phosphatase,glutamic-pyruvic transaminase(GPT),glutamic-oxaloacetic transaminase(GOT),albumin(ALB),lactate dehydrogenase,uric acid,urea,and creatinine were detected.Results The levels of GPT and GOT in the peripheral blood of metacestode of Echinococcus multilocularis infection group were higher than those of control group,and the differences were statistically significant(P<0.05 or P<0.01).The levels of GPT and GOT in peripheral blood of NK cell clearance group were higher than those of NK cell clearance control group,and TP and ALB were lower than those of NK cell clearance control group,the differences were statistically significant(P<0.05 or P<0.01).The level of TP inperipheral blood of CD8^(+)T cell clearance group was lower than that of CD8^(+)T cell clearance control group,and the difference was statistically significant(P<0.05).Conclusion Metacestode of Echinococcus multilocularis infection can cause liver function damage to a certain extent,especially in the state of NK cell depletion.