人牙龈间充质干细胞对B细胞的作用及机制研究  

Effect and mechanism of human gingival mesenchymal stem cell on B cells

作　　者：张楷 陈柯妍 李凯[3] 马铁梁 古鉴[4] Zhang Kai;Chen Keyan;Li Kai;Ma Tieliang;Gu Jian(Department of Hepatobiliary and Laparoscopic Surgery,Yixing Hospital affiliated to Jiangsu University,Yixing 214200,China)

机构地区：[1]江苏大学附属宜兴医院肝胆、腹腔镜外科,江苏宜兴214200 [2]江苏大学附属宜兴医院实验室,江苏宜兴214200 [3]江苏大学 [4]南京医科大学第一附属医院肝脏外科

出　　处：《器官移植》2020年第2期253-258,共6页Organ Transplantation

基　　金：江苏省2013年度“六大人才高峰”高层次人才资助项目(2013-WSN-025);宜兴市科技创新专项资金项目(2017SF06)。

摘　　要：目的探讨人牙龈间充质干细胞(GMSC)对B细胞增殖与分化的调节功能及其相关分子机制。方法分离提取GMSC,从外周血分选出B细胞。将GMSC或纤维母细胞分别与B细胞进行体外共培养,设为GMSC组和纤维母细胞组,检测两组B细胞的扩增情况;检测两组细胞上清中IgG1与IgM的表达情况;检测两组细胞白细胞介素(IL)-6、Perforin、干扰素(IFN)-γ和肿瘤坏死因子(TNF)-α的分泌情况;检测两组B细胞表达IL-10与转化生长因子(TGF)-β的水平;检测两组B细胞中PC-1的表达情况;检测GMSC调控B细胞功能的信号通路;检测GMSC对B细胞活化T细胞功能的调控。结果与纤维母细胞组比较,GMSC组B细胞的扩增明显减弱(P<0.05),GMSC与B细胞共培养后,能明显抑制B细胞分泌IgG1和IgM,明显抑制IL-6、Perforin、IFN-γ和TNF-α的分泌(均为P<0.05)。与纤维母细胞组比较,GMSC组中的IL-10与TGF-β分泌量更高(均为P<0.05)。GMSC组中PC-1的表达水平明显降低(P<0.05)。加入TGF-β受体抑制剂ALK5后,GMSC对B细胞的抑制作用明显减弱(P<0.05)。与纤维母细胞组比较,GMSC组的B细胞活化和扩增T细胞的能力明显减弱(P<0.05)。结论GMSC对B细胞及其介导的免疫反应具有抑制功能,可通过TGF-β信号通路抑制B细胞的活化以及其他相关功能。Objective To investigate the regulating function of human gingival mesenchymal stem cell(GMSC)on the proliferation and differentiation of B cells and its underlying molecular mechanism.Methods GMSC were isolated and B cells were isolated from peripheral blood.GMSC or fibroblasts were co-cultured with B cells in vitro and assigned into the GMSC group and fibroblast group.The proliferation of B cells was detected in two groups.The expression of IgG1 and IgM in the cell supernatants was measured between two groups.The secretion of interleukin(IL)-6,Perforin,interferon(IFN)-γand tumor necrosis factor(TNF)-αwas compared between two groups.The expression levels of IL-10 and transforming growth factor(TGF)-βin B cells were detected between two groups.The expression of PC-1 in B cells was measured in two groups.The signaling pathway involved with the regulating effect of GMSC on B cell function was investigated.The regulating effect of GMSC on the role of B cells in activating T cell function was assessed.Results Compared with the fibroblast group,the proliferation of B cells was significantly weakened in the GMSC group(P<0.05).Co-culture of GMSC and B cells significantly inhibited the secretion of IgG1 and IgM from B cells and the secretion of IL-6,Perforin,IFN-γand TNF-α(all P<0.05).Compared with the fibroblast group,the secretion of IL-10 and TGF-βwas significantly higher in the GMSC group(both P<0.05).The expression level of PC-1 in the GMSC group was significantly down-regulated(P<0.05).After adding ALK5,an inhibitor of TGF-βreceptor,the inhibitory effect of GMSC upon B cells was significantly weakened(P<0.05).Compared with the fibroblast group,the ability of B cells to activate and proliferate T cells was significantly attenuated in the GMSC group(P<0.05).Conclusions GMSC can inhibit B cells and their mediated immune responses.The activation of B cells and other related functions can be suppressed through the TGF-βsignaling pathway.

关 键 词：移植免疫 慢性移植物抗宿主病 牙龈间充质干细胞 人外周血单核细胞 B细胞 T细胞 调节性B细胞 转化生长因子 

分 类 号：R617[医药卫生—外科学] R392.4[医药卫生—临床医学]