Carboxymethylpachymaran Enhances Immunologic Function of Dendritic Cells Cultured in Two Kinds of Hepatoma Carcinoma Cell Line's Supernatant via Nuclear Factor κ B/Rel Pathway  

作　　者：陈卓 于斌 吴先林 戴聪奇 钱国强 于建中 何海滨 王志新 侯军 陈孝银 

机构地区：[1]Medical College Jinan University [2]Nursing Department Huanghuai University [3]Tong rentang Hospital [4]Heshan People Hospital [5]Dongpu Branch of the First Affiliated Hospital of Jinan University

出　　处：《Chinese Journal of Integrative Medicine》2012年第3期203-208,共6页中国结合医学杂志（英文版）

基　　金：Supported by the National Natural Science Foundation of China (No.30973693);the Research Fund for the Doctoral Program of Higher Education of China(No.20104401110003)

摘　　要：Objective：To study the immunologic function of dendritic cells（DCs） cultured in two kinds of hepatoma cell line＇s supernatant and the enhancing effects of carboxymethylpachymaran（CMP） on DCs.Methods：DCs were harvested after stimulation by granulocyte-macrophage colony-stimulating factor （GM-CSF） and interleukin（IL）-4 from umbilical cord blood using density-gradient centrifugation method. Cultured supernatant of two hepatoma cell lines（HepG2 and HepG2.2.15） were collected for condition medium（CM） according to a volume ratio of supernatant to incomplete RPMI-1640 medium,which was 3：1.CMP was dissolved in incomplete RPMI-1640 medium.Experimental groups were divided according to the culture medium,either CM or with CMP in it.DCs subsets CD83,CD86,CD1a,and d-related human leukocyte antigens（HLA-DR） were analyzed by flow cytometry.The proliferation ability of allogeneic T cells in mixed lymphocyte reaction（MLR） stimulated by DCs was examined by 3-（4,5-dimethylthiazol-2-yl）-2,5- diphenyltetrazolium bromide（MTT） analysis.IL-12p70,interferon-γ（IFN-γ）,and nuclear factor k B（NF- k B） were detected by enzyme-linked immunosorbent assay analysis.Results：The proliferation of lymphocytes and secreting level of IL-12 and expression of phenotype of DCs cultured in two kinds of CM were lower than those of normal group（P0.01）.Compared with the normal group,groups treated with CMP showed a higher expression level of DCs subsets,lymphocyte reproductive activity,as well as IL-12 and IFN-γsecretion levels.Groups treated with CMP also demonstrated higher levels of DCs phenotype expression and IL-12 and IFN-γsecretion in supernatant of MLR and higher lymphocyte reproductive activity compared with CM group（P0.05）.Compared with the normal group,the expression level of NF-k B in DCs nuclear was higher in CMP groups but lower in two CM groups（P0.05）.After CMP was added,the NF-κB expression levels of two CM groups were increased compared with levels before CMP was addedObjective：To study the immunologic function of dendritic cells（DCs） cultured in two kinds of hepatoma cell line＇s supernatant and the enhancing effects of carboxymethylpachymaran（CMP） on DCs.Methods：DCs were harvested after stimulation by granulocyte-macrophage colony-stimulating factor （GM-CSF） and interleukin（IL）-4 from umbilical cord blood using density-gradient centrifugation method. Cultured supernatant of two hepatoma cell lines（HepG2 and HepG2.2.15） were collected for condition medium（CM） according to a volume ratio of supernatant to incomplete RPMI-1640 medium,which was 3：1.CMP was dissolved in incomplete RPMI-1640 medium.Experimental groups were divided according to the culture medium,either CM or with CMP in it.DCs subsets CD83,CD86,CD1a,and d-related human leukocyte antigens（HLA-DR） were analyzed by flow cytometry.The proliferation ability of allogeneic T cells in mixed lymphocyte reaction（MLR） stimulated by DCs was examined by 3-（4,5-dimethylthiazol-2-yl）-2,5- diphenyltetrazolium bromide（MTT） analysis.IL-12p70,interferon-γ（IFN-γ）,and nuclear factor k B（NF- k B） were detected by enzyme-linked immunosorbent assay analysis.Results：The proliferation of lymphocytes and secreting level of IL-12 and expression of phenotype of DCs cultured in two kinds of CM were lower than those of normal group（P0.01）.Compared with the normal group,groups treated with CMP showed a higher expression level of DCs subsets,lymphocyte reproductive activity,as well as IL-12 and IFN-γsecretion levels.Groups treated with CMP also demonstrated higher levels of DCs phenotype expression and IL-12 and IFN-γsecretion in supernatant of MLR and higher lymphocyte reproductive activity compared with CM group（P0.05）.Compared with the normal group,the expression level of NF-k B in DCs nuclear was higher in CMP groups but lower in two CM groups（P0.05）.After CMP was added,the NF-κB expression levels of two CM groups were increased compared with levels before CMP was added

关 键 词：dendritic cells carboxymethylpachymaran HEPATOCARCINOMA lymphocyte subsets mixed leukocyte reaction 

分 类 号：R735.7[医药卫生—肿瘤]