摘要
目的了解谷氨酸钠致单离培养的小鼠耳蜗螺旋神经节细胞损伤后,碱性成纤维细胞生长因子(bFGF)对其生存及生长的影响,以及二者对细胞内游离钙离子浓度的影响。方法在培养液中加入2×102mol/L的谷氨酸钠2小时后,实验组分别换成含bFGF25、50、100μg/L的培养液,对照组加空白培养液。结果24小时后可见部分细胞裂解、死亡,对照组明显多于实验组。培养14天后甲苯胺蓝染色证实,实验组的细胞生存数及细胞突起长度明显高于对照组,差异有显著性(F检验,P<0.01),且随着bFGF浓度增加,细胞生存数及细胞突起长度均增加,不同浓度实验组间差异有显著性(P<0.01)。用Fluo3染色,激光扫描共聚焦显微镜观察10个螺旋神经节细胞,培养液中加入谷氨酸钠后细胞内钙离子浓度迅速上升,在10~15秒内达峰值,100秒左右恢复到正常水平;加入bFGF后细胞内钙离子浓度无明显变化(t检验,P=0.204)。结论谷氨酸钠可致螺旋神经节细胞内钙离子浓度升高,bFGF可以降低谷氨酸钠对螺旋神经节细胞的细胞毒性。
Objective To understand the effects of basic fibroblast growth factor (bFGF) on survival and neurogenesis of the cultured spiral ganglion cell (SGC) of mouse after exposed to glutamate, and the effects of bFGF and glutamate on free intracellular calcium ion concentration. Methods After exposed to 2×10 -2 mol/L glutamate for 2 hours , the medium of SGC was replaced with the media containing 25,50, 100 μg/L bFGF. Medium without bFGF was used as control. Results Twenty four hours later, more cells in the control group showed signs of cell death than in bFGF groups. After 14 days, specimens were fixed in 4% paraformaldehyde and stained with toluidine blue. It was shown that the SGC in medium with bFGF exhibited more survival and longer neurites, and the effect was dose dependent ( P <0.01). Application of glutamate to the medium induced an increase to the the free intracellular calcium ion concentration in SGC, but bFGF had no such effect. Conclusion Glutamate excitotoxicity was associated with free intracellular calcium ion concentration elevation in SGC, and bFGF could protect SGC against glutamate neurotoxicity.
出处
《中华耳鼻咽喉科杂志》
CSCD
1998年第3期165-168,共4页
Chinese Journal of Otorhinolaryngology
基金
国家自然科学基金
关键词
耳蜗
螺旋神经节
成纤维细胞
生长因子
谷氨酸钠
Cochlea
Spiral ganglion
Fibroblast growth factor
basic
Sodium glutamate