摘要
目的探讨多房棘球绦虫原头蚴对体外培养宿主肝细胞丝裂素活化蛋白激酶(mitogen activated protein kinase,MAPK)信号通路的影响。方法采用宿主大鼠肝细胞BRL3A和人源肝癌细胞系HepG2,分别与多房棘球绦虫原头蚴体外无血清共培养。实验组(与多房棘球绦虫原头蚴共培养)每瓶细胞接种约3×103个原头蚴;处理组(U0126)每瓶细胞先用20μmol/L U0126处理3 h,再接种约3×103个原头蚴,继续共培养,分别在12、24、48、72、96和120 h收集细胞,同时收集对照组(无血清培养基)细胞。利用免疫印迹技术(Western blot)检测p-ERK1/2、p-p38和p-JNK的变化。结果在48 h,肝细胞BRL3A共培养组p-ERK1/2表达活性与对照组相比差异有统计学意义(P<0.05),在12、24和48 h,p-JNK和p-p38表达活性分别有微弱升高;在72和96 h肝细胞HepG2共培养组p-ERK1/2表达活性与对照组相比差异有统计学意义(P<0.05),p-JNK和p-p38表达活性无明显变化;U0126处理组p-ERK1/2活性受抑制。结论多房棘球绦虫原头蚴可能自身分泌细胞因子EmIns、EmBMP1/2、EmAct或egfd,通过与宿主表面受体结合激活宿主肝细胞MAPK信号通路。
Objective To explore the effect of Echinococcus multilocularis protoscoleces on the activation of host hepatocyte MAPK（mitogen-activated protein kinase） signaling pathways.Methods BRL3A rat hepatocytes and HepG2 human hepatocellular liver carcinoma cells were dispensed into 25 cm2 culture flasks for co-culturing.The experimental group（treated with protoscoleces）： BRL3A or HepG2 cells incubated with almost 3 000 protoscoleces for 12,24,48,72,96 and 120 h.The experimental group（treated with U0126）： BRL3A or HepG2 cells were pretreated with 20 μmol/L U0126 for 3 h and then infected with E.multilocularis.The cells were harvested 12,24,48,72 96 and 120 h postinfection and subjected to Western blot.Results As a result of co-culturing with BRL3A,the expression of p-ERK significantly increased at 48 h（P0.05） and the expression of p-JNK and p-p38 increased slightly at 12,24,and 48 h according to Western blot.As a result of co-culturing with HepG2,the expression of p-ERK significantly increased at 72 and 96 h while the expression of p-JNK and p-p38 did not change.The expression of p-ERK was inhibited by treatment with U0126.Conclusion E.multilocularis protoscolex-derived cytokines and hormones such as EmIns,EmBMP1/2,EmAct,and egfd may bind to corresponding host hepatocyte receptors,thus affecting the activation of MAPK signaling pathways in vitro.
出处
《中国病原生物学杂志》
CSCD
2011年第8期574-577,608,共5页
Journal of Pathogen Biology
基金
国家自然科学基金项目(No.30860253)
新疆包虫病基础医学重点实验室开放课题资助项目(No.XJDX0202-2009-01)
