【摘要】 目的考察灯盏细辛中黄酮类成分(总黄酮、灯盏花素)对体外高压诱导的视网膜神经节细胞(retinal gan-glion cells,RGCs)凋亡的影响,探讨其视神经保护作用的物质基础。方法采用胰酶消化法将24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)包被的盖玻片中。培养72h后,将覆有细胞的血盖片转入加压装置中,加入黄酮类成分,继续培养48h,采用Fas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Fas蛋白阳性表达指数和凋亡指数均明显低于模型组(P<0.05~0.01)。结论黄酮类成分均能对抗压力诱导的RGCs凋亡,为灯盏细辛视神经保护的有效组分。更多还原

【Abstract】 Objective To observe the effects of the flavonoids on apoptosis of retinal ganglion cells by high intraocular pressure in vitro and to explore the substantial foundation with neuroprotective effects in glaucoma of E.breviscapus.Methods The retina of 24 Sprague-Dawley rats born only for 2 to 3 days 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 cultured for 72 hours,glass cover were put into the high intraocular pressure device and the flavonoids were added to the dying vats and then culture was kept on for 48 hours.The expression of Fas protein was observed 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% of the living cells were retinal ganglion cells by NSE identification.The positive myocytes of Fas protein and the apoptosis index in the experimental were lower than those in the model group(P<0.05～0.01).Conclusion The flavonoids can protect retinal ganglion cells against apoptosis by high intraocular pressure,which are the main active components of E.breviscapus with neuroprotective effects.更多还原