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目的 从信号分子PKC和细胞内游离Ca2 + ([Ca2 + ]i)这一途径探索青春型双歧杆菌的脂磷壁酸 (lipoteichoicacid ,LTA)激活小鼠腹腔巨噬细胞的机制 ,同时观察巨噬细胞之间缝隙连接通讯(GJIC)的变化。方法 γ 3 2 PATP磷酸转移法测定巨噬细胞总PKC活性 ;激光共聚焦显微镜检测 [Ca2 + ]i浓度的变化 ;激光漂白后荧光恢复技术 (FRAP)观察GJIC的状态。结果 (1)LTA能明显提高巨噬细胞总PKC活性 ,呈剂量依赖性 ;(2 )LTA可显著升高巨噬细胞 [Ca2 + ]i的水平 ,并且EDTA和维拉帕米处理组、肝素和普鲁卡因处理组以及EDTA、维拉帕米、肝素和普鲁卡因处理组巨噬细胞内 [Ca2 + ]i亦升高 ,但明显低于对照组 (P <0 .0 1)。 (3)巨噬细胞被LTA刺激后 ,其平均荧光恢复率明显高于对照组(P <0 .0 1)。结论 LTA可通过提高PKC活性 ,升高 [Ca2 + ]i的水平以及增强GJIC的功能来激活巨噬细胞
OBJECTIVE: To explore the mechanism of lipoteichoic acid (LTA) activation in mouse peritoneal macrophages from Bifidobacterium adolescentis via PKC signaling pathway and intracellular free Ca2 + ([Ca2 +] i) pathway. Changes in gap junctional communication (GJIC) between macrophages. Methods The total PKC activity of macrophages was measured by γ 3 2 PATP phosphate transfer assay. The changes of [Ca2 +] i concentration were detected by laser scanning confocal microscopy. The state of GJIC was observed by fluorescence recovery after laser bleaching (FRAP). Results (1) LTA significantly increased the total PKC activity of macrophages in a dose-dependent manner. (2) LTA increased the [Ca2 +] i level of macrophages, and the effect of EDTA and verapamil on heparin And [Ca2 +] i in macrophages of treated group, procaine and EDTA, verapamil, heparin and procaine increased, but were significantly lower than that of control group (P <0.01) . (3) After macrophages were stimulated by LTA, the average fluorescence recovery rate was significantly higher than the control group (P <0.01). Conclusion LTA can activate macrophages by increasing the activity of PKC, increasing the level of [Ca2 +] i and enhancing the function of GJIC