灯盏花素对体外高压致视网膜神经节细胞凋亡的影响  

Effects of Breviscapine on Apoptosis of Retinal Ganglion Cells by High Intraocular Pressure in Vitro

作　　者：盛艳梅[1] 孟宪丽[2] 张艺[2] 龙怡[2] 

机构地区：[1]成都医学院药理教研室,成都610083 [2]成都中医药大学药学院,成都610075

出　　处：《成都医学院学报》2007年第1期26-29,共4页Journal of Chengdu Medical College

基　　金：国家自然科学基金资助项目(NO.30271581)

摘　　要：目的考察灯盏花素对体外高压诱导的视网膜神经节细胞(retinal ganglion cells,RGCs)凋亡的影响,探讨灯盏细辛视神经保护作用的物质基础。方法用胰酶消化法将20只出生2～3d的SD(Sprague-Dawley)乳鼠视网膜制成细胞悬液,接种于经多聚鸟氨酸(HA)和层粘连蛋白(LN)包被的血盖片中。培养72h后,将覆有细胞的血盖片转入加压装置中,加入灯盏花素溶液,继续培养24、48h,采用Caspase-3蛋白免疫组化染色法及原位末端脱氧核苷酸转移酶标记(Tunel-POD)法进行检测,每天观察细胞形态,同时对部分细胞行NSE染色检查。结果细胞生长良好,NSE染色表明,85%以上的细胞为RGCs。给药组的Caspase-3蛋白阳性表达指数和凋亡指数均明显低于模型组(P<0·05,P<0·01)。结论灯盏花素能对抗压力诱导的RGCs凋亡,为灯盏细辛视神经保护的有效组分。Objective To investigate the effects of Breviscapine on apoptosis of retinal ganglion cells by high intraocular pressure in vitro and explore the substantial foundation with neuroprotective effects in glaucoma of E. breviscapus. Methods The retinal of 20 post- natal 2- 3 days Sprague- Dawley rats were dissociated into cell suspension with trypsin digestion. The cell suspension was implated in 6 - well culture plates covered with hyaluronic acid and laminin in each well. After culturing for 72 hours, glass cover were putted into the high intraocular pressure device and the Breviscapine were added to the dyeing vats, continue to culture 24/48 hours. Observing the expression of Caspase - 3 protein by the immuneohistochemistry method and apoptotic cells were detected by TUNEL - POD method. And some of the 5 - day culture cells were identified by NSE technique. Results The cells grew very well, over 85 percent of the hying cells were retinal ganglion cells by NSE identification. The positive myocytes of Caspase - 3 protein and the apoptosis index in the experiment were significantly lower than those in the model group （ P 〈 0.05, P 〈 0.01）. Conclusions Breviscapine can protect retinal ganglion cells against apoptosis by high intraocular pressure, Which is the main active component of E. breviscapus with neuroprotective effects.

关 键 词：灯盏花素 视网膜神经节细胞培养 高眼压模型 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]