压力对大鼠纯化视网膜神经节细胞中bcl-2、bax及p53mRNA表达的影响  

INFLUENCE OF PRESSURE ON mRNA EXPRESSION OF BCL-2, BAX AND p53 IN PURIFIED RETINAL GANGLION CELLS OF RATS

作　　者：胡竹林[1] 杜蜀华[2] 

机构地区：[1]云南省红十字会医院眼科,昆明650021 [2]华中科技大学同济医学院附属同济医院眼科,武汉430030

出　　处：《中国现代医学杂志》2002年第19期24-27,30,共5页China Journal of Modern Medicine

摘　　要：目的 :通过检测不同压力下纯化培养的大鼠视网膜神经节细胞 (Retinalganglioncells,RGCs)中bcl - 2 ,bax ,p5 3mRNA表达情况 ,了解压力对视网膜神经节细胞凋亡的影响。方法 :用SD系大鼠脑源性星型胶质细胞同化培养液双界面法[1] 培养羊抗鼠Thy1.1单克隆抗体纯化的SD系大鼠RGCs;在纯化培养的RGCs上分别施加 0 ,2 0mmHg ,4 0mmHg ,6 0mmHg和 80mmHg气压 ,培养 2 4h及 4 8h后TUNEL法检测各组细胞的凋亡 ,原位杂交检测细胞中bcl- 2 ,bax及p5 3mRNA的表达。结果 :纯化的RGCs培养 12h及 2 4h用羊抗鼠FITC -Thy - 1.1单克隆抗体进行鉴定阳性 ,纯度为 97%。传三代后星型胶质细胞用胶质纤维酸性蛋白(GFAP)单克隆抗体标记和鉴定阳性。培养 2 4h组TUNEL法检测对照组极少数细胞凋亡 ,2 0mmHg组见较多细胞凋亡 ,随着压力的增高细胞凋亡数增多 ,凋亡指数增高 ;与对照组相比差异均有显著性。培养 4 8h组压力≥ 2 0mmHg各组较相同压力培养 2 4h组凋亡指数增高 ,差异有显著性 (P <0 .0 5 )。压力≥ 4 0mmHg时bcl-2mRNA表达逐渐减少 ,压力≥ 2 0mmHg时bax及 p5 3mRNA表达则逐渐增加 ;与对照组相比均有显著性差异 (P <0 .0 5 )。结论 :高于 2 0mmHg压力时间超过 2 4h可激活体外培养的视网膜神经节细胞的相关凋亡基因导致细?Objective:To study the bcl-2, bax and p53 mRNA expression in rat purified retinal ganglion cells(RGCs) that cultured in different pressures.Method:To culture RGCs that from Sprague Dawley(SD) neonatal rats(postnatal 1～5 day) on two planes in assimilative culture solution, RGCs were purified by Thy1.1 with sheep anti rat FITC monoclonal antibody. RGCs were cultured under pressures of 0mmHg, 20mmHg, 40mmHg, 60mmHg and 80mmHg for 24 hours and 48 hours, respectively. The changes of bcl-2, bax and p53 mRNA in RGCs under different pressures were demonstrated qualitatively and quantitatively by in situ hybridization and their apoptosis were detected by TUNEL method, respectively.Results:After cultured 12 and 24 hours in vitro, the purification rate of RGCs in the experiment arrived at 98 percent. After cultured 24 hours, the expression of apoptotic cells in RGCs became higher and higher as the pressure was increased. There was a few apoptotic cells expression in the control group (0mmHg) and 20mmHg, 40mmHg, 60mmHg and 80mmHg groups was significantly different from the control group,respectively. ( P <0.05). After cultured 48 hours, 20mmHg, 40mmHg,60mmHg and 80mmHg groups was significantly different from which of cultured 24 hours, respectively ( P <0.05). If the pressure was equal to or higher than 40mmHg, the expression of bcl-2 mRNA in RGCs became weaker and weaker as the pressure was increased,and was significantly different from control group, respectively( P <0.05). If the pressure was equal to or higher than 20mmHg, the expression of bax and p53 mRNA in RGCs became higher and higher as the pressure was increased,and was significantly different from control group, respectively( P <0.05).Conclusion: Pressure can cause the expression of bcl-2 to be weaker and bax, p53 mRNA to be higher in purified retinal ganglion cells in vitro and induce or aggravate RGCs to apoptosis.

关 键 词：细胞凋亡 压力 大鼠 视网膜神经节细胞 BCL-2 BAX p53mRNA表达 

分 类 号：R774[医药卫生—眼科]