论文部分内容阅读
目的通过观察石斛散对人视网膜神经细胞内钙离子持续升高导致的细胞凋亡的保护作用,探讨其在治疗视网膜变性类疾病方面的作用及机制,为临床治疗同类疾病提供药理学依据。设计实验性研究。研究对象体外培养的人眼视网膜神经细胞。方法利用体外培养的贴壁、伸展以及生长良好人视网膜神经细胞,用Fluo3／AM标记,通过激光共焦扫描显微镜动态观察和记录石斛散和维拉帕米对细胞内钙离子水平以及谷氨酸引起的钙超载的影响。主要指标细胞内钙离子水平(荧光强度)。结果加入谷氨酸后,荧光强度较原基础值增加88％;而石斛散可使荧光强度降低14．8％;维拉帕米降低幅度为57．3％。对先用谷氨酸干预的细胞,石斛散不能降低胞内钙离子水平,但预先用石斛散干预的细胞,谷氨酸升高胞内钙离子30％,与未干预组比较有统计学差异(P=O．000)。结论石斛散具有降低体外培养人视网膜神经细胞内钙离子的作用,同时可以抵抗谷氨酸损伤细胞后引起的胞内钙离子升高,提示其具有抵抗钙超载,抑制视网膜细胞凋亡的作用。
Objective To observe the protective effect of Shiqi Powder on apoptosis induced by sustained increase of intracellular calcium in human retinal nerve cells, to explore its role and mechanism in the treatment of retinal degenerative diseases, and to provide pharmacological basis for clinical treatment of similar diseases. Design experimental studies. The subjects were cultured in vitro with human eye retinal neurons. METHODS: In vitro cultured adherent, stretched, and well-developed human retinal neurons were stained with Fluo3/AM to observe and record the intracellular calcium levels and glutamate dynamics by laser confocal scanning microscopy. Causing calcium overload effects. The main indicators of intracellular calcium levels (fluorescence intensity). Results After adding glutamic acid, the fluorescence intensity increased by 88% compared with the original baseline value; while Shiqisan decreased the fluorescence intensity by 14.8% and verapamil decreased by 57.3%. For cells first treated with glutamate, Shiqisan could not reduce intracellular calcium levels, but glutamate increased the intracellular calcium by 30% in cells pretreated with Shiqisan, which was statistically different from the non-intervention group. (P=O.000). Conclusion Shiqisan can reduce the intracellular calcium ion of human retinal neurons in vitro, and it can also resist the increase of intracellular calcium induced by glutamate-injured cells, suggesting that it has the function of resisting calcium overload and inhibiting apoptosis of retina cells.