碱性成纤维细胞生长因子作用于人晶状体上皮细胞系内Ca2+转导通路的初步研究

目的 初步探讨碱性成纤维细胞生长因子(bFGF)作用于人晶状体上皮细胞(LEC)系-B3(LEC-B3)内Ca^2 转导的途径。方法 人LEC-B3传代培养,在激光扫描共焦显微镜下于解冻后的第三代细胞中分别加入10μg／L bFGF、10mmol／L咖啡因、10mmol／L兰尼碱、50mmol／L普鲁卡因及0．5mmol／L金雀异黄素,通过观测细胞内相对荧光强度实时观察细胞内Ca^2 浓度的变化。结果 10μg/L bFGF在细胞外液含或无Ca^2 、Mg^2 的情况下,均可迅速引起人LEC-B3内Ca^2 浓度升高,且持续时间基本相同;而在细胞外液无Ca^2 和Mg^2 、含1mmol／L乙二醇双乙醚四乙酸时,人LEC-B3内Ca2^ 浓度更高。50mmol／L普鲁卡因和10mmol／L兰尼碱与10μg/L bFGF共同作用,人LEC-B3内Ca^2 浓度虽升高,但低于10μg/L bFGF单独作用的效果;10mmol／L咖啡因和0．5mmol／L金雀异黄素可明显降低追加的10μg/L bFGF升高人LEC-B3内Ca^2 浓度的作用,后者尤为明显。结论 bFGF主要通过激活络氨酸蛋白激酶信号转导途径引起人LEC-B3内Ca^2 浓度升高;肌醇1,4,5-三磷酸受体和兰尼碱受体通道发挥一定作用;Ca^2 主要来源于细胞内Ca^2 库释放。（中华眼科杂志,2004,40：832-835)

The pathway of bFGF induced changes of calcium in cultured human lens epithelial cells

OBJECTIVE: To investigate the pathway of basic fibroblast growth factor (bFGF) induced changes of calcium in a human lens epithelial cell line LEC-B3. METHODS: The 3rd passage of LEC-B3 cells was used in the present studies. bFGF (10 microg/L), caffeine (10 mmol/L), ryanodine (10 mmol/L), procaine (50 mmol/L) and genistein (0.5 mmol/L) were added to the culture medium. Changes of calcium concentration were observed and analyzed with laser scanning confocal microscope. RESULTS: bFGF (10 microg/L) could induce a prompt increase of calcium concentration when cells culturing in culture medium with or without calcium. No obvious difference of calcium concentration could be observed between these two conditions except that the amplitude of increase of calcium concentration under calcium free solutions was slightly higher than that in medium with calcium. Both 50 mmol/L procaine and 10 mmol/L ryanodine partially inhibited the calcium increase induced by bFGF. After the addition of 10 mmol/L caffeine, 10 microg/L bFGF could not induce a significant increase of calcium. Genistein at 0.5 mmol/L significantly inhibited bFGF induced changes of calcium. CONCLUSIONS: bFGF inducing calcium ion concentration increase mainly depends on TPK pathway. Most of the calcium ions come from the release of intracellular calcium pools in human lens epithelial cell. Both the IP(3R) and RyR take part in the action, while IP(3R) is the more important factor.