摘要
目的探讨丙型肝炎病毒(hepatitis C virus,HCV)F蛋白对白介素13(interluekin-13,IL-13)表达的影响及其相关机制。方法收集慢性HCV1b型患者(chronic hepatitis patient,CHP)的血样,检测F抗体(F antibody,FAb)的阳性率并分成2组(F-Ab(+)CHP、F-Ab(-)CHP);收集健康者血样作为对照组;用酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测各组样本血清中IL-13的水平;分离患者外周血单个核细胞(peripheral blood mononuclear cells,PBMC),用不同浓度的F蛋白刺激72 h后,ELISA法检测细胞上清中IL-13的水平,蛋白质免疫印迹(western blotting)法检测细胞中GATA结合蛋白3(GATA binding protein 3,GATA-3)和活化T细胞核因子(nuclear factor of activated T cell,NFAT1)蛋白表达水平。结果 F-Ab(+)CHP组血清IL-13水平高于F-Ab(-)CHP组(P<0.001);PBMC经F蛋白体外刺激后,细胞上清中IL-13水平随着F蛋白刺激浓度的增高而增高(x2=13.52,P=0.009);PBMC中GATA-3蛋白表达水平不受F蛋白刺激的影响(均有P>0.05),而F蛋白可下调NFAT1蛋白表达。结论在体外培养的PBMC中,F蛋白有上调IL-13水平的作用,而且可能通过下调NFAT1间接发挥作用。
Objective To investigate the effect of hepatitis C virus F protein on interluekin-13 and related mecha- nism. Methods Blood samples were collected from chronic HCV-lb patients （CHP）, and divided into F-Ab（ ＋ ）CHP group and F-Ab（ - ） CHP group, while blood samples from healthy persons were used as controls. The levels of IL-13 in all subjects＇ sera were determined by enzyme linked immunosorbent assay （ELISA）. Isolated peripheral blood mononuclear cells （PBMC） were stimulated by different concentrations of F protein for 72 hours, then cells were collected to test the level of GATA binding protein 3 （ GATA-3 ） and nuclear factor of activated T eel1 （ NFAT1 ） by Western blotting. And the cell supernatants and blood sera were collected to determine the IL-13 level by ELISA. Results The level of IL-13 in F- Ab （ ＋ ） CHP group was significantly higher than that in F-Ab （ - ） CHP group （P 〈0. 001 ） ; IL-13 level in cell superna- tant was up-regulated along with increased concentrations of F protein （X2 = 13.52 ,P =0. 009）. The expression of GATA- 3 was not affected by F protein in vitro （P 〉 0. 05 ）, while NFAT1 was down-regulated by F protein. Conclusions HCVlb F protein can up-regulate the expression of IL-13 in vitro, which may attribute to the indirect effect of NFAT1 down-regula-
出处
《中华疾病控制杂志》
CAS
CSCD
北大核心
2015年第2期115-118,126,共5页
Chinese Journal of Disease Control & Prevention
基金
国家自然科学基金(81172724
30972628)
十二五国家科技重大专项(2011ZX10004-902)
江苏省自然科学基金面上项目(BK2010112
BK2011840)
