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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。 Abstract: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)-γ and tumor necrosis factor (TNF)-α was compared between two groups. The expression levels ofIL-10 and transforming growth factor (TGF)-β in 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 ofIL-6, Perforin, IFN-γ and 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-β receptor, 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-β signaling pathway. -
图 2 GMSC对B细胞扩增的调节情况
注：与纤维母细胞组比较，aP < 0.05。
Figure 2. Regulation of GMSC in B cell proliferation
图 3 GMSC对B细胞分泌功能的调节情况
注：A图示B细胞分泌IgG1的情况；B图示B细胞分泌IgM的情况；C图示B细胞分泌细胞因子的情况；与纤维母细胞组比较，aP < 0.05。
Figure 3. Regulation of GMSC in B cell secretion function
图 4 GMSC对B细胞分化的调节情况
注：A图示B细胞表达IL-10与TGF-β的水平，与纤维母细胞组比较，aP < 0.05；B图示B细胞中PC-1的表达情况。
Figure 4. Regulation of GMSC in B cell differentiation
图 5 ALK5在GMSC调节B细胞中的作用
Figure 5. The effect of ALK5 on the regulation of B cell by GMSC
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