目的评价重明益损汤药物血清抗视网膜神经节细胞凋亡的作用。方法体外培养视网膜神经节细胞(retinal ganglion cells,RGCs),采用谷氨酸作为细胞凋亡诱导剂建立RGCs凋亡模型,并用重明益损汤药物血清加入细胞培养基进行干预,分别用四甲基偶氮唑盐(MTT)法和流式细胞检测的方法进行细胞存活率及细胞凋亡相关检测。结果终浓度为3.2 mmol/L的谷氨酸作用18 h可造成适量的RGCs凋亡,以此方法制作RGCs凋亡模型。MTT及流式细胞检测示重明益损汤药物血清干预能显著提高谷氨酸损伤后RGCs存活率(P〈0.01),对抗RGCs凋亡,并可下调Bax蛋白的表达。结论重明益损汤药物血清对RGCs具有一定保护作用。
OBJECTIVE To evaluate the anti-apoptosis effect of Chongming Yisun Decoction on cultured retinal ganglion cells(RGCs).METHODS We collected serum from rats which were intragastric administrated with Chongming Yisun Decoction and used it as part of medium with DMEM-F12 in RGCs culture.MTT cell viability assay and flow cytometry were used to calculate survival and apoptosis ratio of RGCs.RESULTS L-glutamate(3.2 mmol/L)could markedly reduce RGCs survival ratio when added to DMEM-F12 medium alone.The collected serum combined with DMEM-F12 could increase survival ratio of RGCs with or without L-glutamate when the concentration of intragastric administrated Chongming Yisun Decoction was 2 g/ml.Our flow cytometry test confirmed that L-glutamate could lead RGCs to apoptosis.We also found that pharmic serum could save RGCs suffered from L-glutamate's and reduce expression of Bax protein.CONLUSIONS Serum collected from rat by intragastric administration of Chongming Yisun Decoction might be a useful drug to protect RGCs.