牛膝多肽对NMDA诱导的大鼠原代视网膜神经节细胞损伤的保护作用

The protective effects of Achyranthes bidentata polypeptides against NMDA-induced cell apoptosis in cultured retinal ganglion cells

目的:探讨牛膝多肽(ABPP)对N-甲基-D-天冬氨酸(NMDA)诱导的体外培养大鼠视网膜神经节细胞(RGCs)损伤的保护作用。方法:采用新生大鼠视网膜神经细胞体外原代培养,随机分为对照组、NMDA组、ABPP+NMDA组[按ABPP不同药物质量浓度(0.1、0.3、1.0、3.0μg/ml)设4个亚组]和地西平(MK-801)+NMDA组。采用倒置显微镜观察细胞形态;抗Thy-1.1细胞免疫荧光染色鉴定培养细胞中RGCs纯度;MTT比色法测定细胞存活率;Hoechst/PI双染色法定性观察细胞凋亡;流式细胞术AnnexinV/PI双染色法定量测定细胞凋亡率。结果:NMDA可诱导大鼠视网膜神经节细胞凋亡,与NMDA组比较,0.3μg/ml ABPP可提高RGCs存活率,减少细胞凋亡(P<0.05)。结论:ABPP可抑制NMDA诱导的视网膜神经节细胞凋亡,保护视网膜神经元。

Objective：To investigate the protective effects of Achyranthes bidentata polypeptides（ABPP） against NMDA-induced apoptosis of cultured rat retinal ganglion cells（RGCs） in vitro.Methods：Neonatal rat retinal neurons were cultrued with or without ABPP.RGCs were first identified immunofluorescently with antibody against Thy1.1.Cell viability was determined by methylthiaz olyltetrazolium chromatometry and apoptosis was examined by annexin V.The effects of ABPP were compared to those of MK-801,a well-known protective against NMDA for retinal ganglion cells.Results：Exposure of RGCs to NMDA may induce neuronal injury.Pretreated with ABPP（0.3 μg/ml） greatly improved the viability of RGCs and attenuated the apoptosis of RGCs.Conclusion：ABPP could inhibit the apoptosis of RGCs induced by exposure to NMDA and provide neuroprotective effects on RGCs.