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目的探讨大鼠小胶质细胞及外周血巨噬细胞对胶质瘤C6细胞株迁移能力的影响。方法将大鼠小胶质细胞及外周血单核细胞诱导分化形成的巨噬细胞分别与胶质瘤C6细胞共培养,以大鼠成纤维细胞共培养为阳性对照,单纯胶质瘤C6细胞为空白对照,共培养0、24、48 h分别行划痕实验,于划痕后6 h观察划痕实验结果;共培养24、48 h时以CD11b标记、流式分选分离小胶质/巨噬细胞及C6细胞,行Western blot检测0、24、48 h小胶质/巨噬细胞IL-10、IL-18和MMP-9及C6细胞株TGF-β、FAS配体表达水平的改变。结果早期小胶质细胞及巨噬细胞均抑制胶质瘤的迁移,巨噬细胞的抑制效果更强(P<0.05);与胶质瘤细胞共培养后两种细胞对肿瘤的抑制作用消失且差异无统计学意义,均表现为促进肿瘤迁移。小胶质细胞、巨噬细胞在初始阶段IL-10、IL-18、金属基质蛋白酶MMP-9表达略有不同,但共培养后两种细胞的上述蛋白表达均升高且一致;共培养后的胶质瘤细胞的TGF-β、FAS配体较共培养前表达均升高且一致。结论小胶质细胞及巨噬细胞与C6胶质瘤细胞共培养后被驯化成同样的GAMs,对C6细胞迁移影响的生物学表现一致。
Objective To investigate the effect of rat microglial cells and peripheral blood macrophages on the migration of C6 glioma cells. Methods The macrophages induced by differentiation of rat microglial cells and peripheral blood mononuclear cells were co-cultured with C6 glioma cells respectively. Co-culture of rat fibroblasts was used as positive control. Glioma C6 cells were Blank control, co-cultured scratch test for 0,24,48 h were scratched 6 hours after the scratch test results were observed; co-cultured 24,48 h marked with CD11b, flow separation of microglia / giant Western blot was used to detect the changes of TGF-β and FAS ligand expression in IL-10, IL-18, MMP-9 and C6 cell lines of microglial / macrophages at 0,24,48 h. Results Both microglial cells and macrophages in early stage inhibited the migration of glioma, and the suppressive effect of macrophages was stronger (P <0.05). After co-cultured with glioma cells, the inhibitory effect of the two cells disappeared The difference was not statistically significant, all showed to promote tumor migration. The expression of IL-10, IL-18 and MMP-9 in microglial cells and macrophages were slightly different at the initial stage, but the expression of the above proteins in both microglial cells and macrophages was increased and consistent after co-culture; Of glioma cells TGF-β, FAS ligand expression before co-culture were elevated and consistent. Conclusions The microglia and macrophages, after being co-cultured with C6 glioma cells, are acclimated to the same GAMs, which has the same biological effect on the migration of C6 cells.