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为观察碱性成纤维细胞生长因子(bFGF)对离体热烫伤后成纤维细胞内钙离子的作用以及丝裂原活化蛋白激酶 (MAPK)通路对胞内游离钙的影响 ,将培养的人成纤维细胞进行热损伤刺激后分成 4组 :①bFGF处理组 (10ng/ml) ;②预先加入PD980 5 9(10 μmol/L)阻断剂 30min,再行bFGF(10ng/ml)刺激 ;③预先加入SB2 0 35 80 (10 μmol/L)阻断剂 30min ,再行bFGF刺激 (10ng/ml) ;④同时加入PD980 5 9(10 μmol/L)和SB2 0 35 80 (10 μmol/L) 2种阻断剂 30min,再加入bFGF(10ng/ml)。应用特异性Ca2 + 荧光指示剂Fluo 3/AM负载细胞 ,激光共聚焦显微镜检测细胞内游离钙的浓度。结果显示 ,受热刺激的成纤维细胞荧光强度较弱。加入bFGF后可促使成纤维细胞中游离Ca2 +浓度升高 ,分别预先加入PD980 5 9和SB2 0 35 80拮抗剂的成纤维细胞 ,再加入bFGF ,胞内钙离子浓度出现不同的钙振荡现象。同时加入 2种阻断剂后胞内钙离子浓度则迅速降低。表明bFGF引起热损伤后的成纤维细胞中游离Ca2 + 浓度的增加 ,MAPK信号通路对胞内钙离子有反馈调节的作用
To observe the effect of basic fibroblast growth factor (bFGF) on intracellular calcium in fibroblasts after ex vivo thermal burns and the effect of mitogen-activated protein kinase (MAPK) pathway on intracellular free calcium, Fibroblasts were divided into 4 groups according to heat injury: ①bFGF treatment group (10ng / ml); ② PD98059 (10μmol / L) blocker pre-added for 30min, then bFGF (10ng / ml) SB203580 (10 μmol / L) blocker for 30 min followed by stimulation with bFGF (10 ng / ml); (4) PD980 5 9 (10 μmol / L) and SB2 0 35 80 Blocker for 30 min followed by addition of bFGF (10 ng / ml). The cells were loaded with Fluo 3 / AM, a specific fluorescent Ca2 + indicator. The concentration of intracellular free calcium was detected by confocal microscopy. The results showed that the heat-stimulated fibroblasts fluorescence intensity is weak. The addition of bFGF increased the concentration of free Ca2 + in fibroblasts. The fibroblasts of PD98059 and SB2 + 3580 antagonist were added beforehand. After bFGF was added, different Ca2 + oscillations appeared in the concentration of intracellular Ca2 +. After adding two kinds of blockers, the intracellular calcium concentration decreased rapidly. Indicating that bFGF causes an increase in free Ca2 + concentration in fibroblasts after heat injury and a feedback-regulated effect of MAPK signaling pathway on intracellular calcium ions