石斛散对体外培养人视网膜细胞的作用

Effects of Shihusan on human retinal cells cultured in vitro

目的观察中药方剂石斛散及单味药对人视网膜细胞的增生以及谷氨酸损伤的保护作用,探讨其在视网膜细胞损伤或变性类疾病方面的作用及机理,为临床治疗同类疾病提供依据。方法体外培养人视网膜细胞,通过四甲基偶氮唑盐实验观察石斛散复方以及各单味药对视网膜细胞增生的影响。另用Rhodamine123标记培养的视网膜细胞,通过激光共聚焦扫描显微镜动态观察和记录不同药物作用下细胞的线粒体膜电位(mitochondrial membrane potential,MMP)变化。结果体外培养人视网膜细胞贴壁、伸展以及生长良好。四甲基偶氮唑盐实验:石斛散组1.58g·L-1和0.50g·L-1以及石斛组5.00g·L-1对人视网膜细胞活性分别增加21.8%、14.2%和13.4%。MMP实验:石斛散轻度升高MMP,增加约13.5%,同时它可以抵抗谷氨酸损伤细胞后引起的MMP下降,由降低46.4%减少为16.2%。结论石斛散能够影响人视网膜细胞线粒体功能、促进细胞活性、抵抗谷氨酸对人视网膜细胞损伤作用。可作为临床治疗视网膜变性类疾病的一种选择。

Objective To explore the effects of Shihusan and single herb on proliferation of human retinal cells and protective function for damage of glutamate,and to investigate its function and mechanisms on retinal damage or degenerated diseases and to offer the basis for the treatment of the similar diseases clinically. Methods Human retinal cells were cultured in vitro. The effects of Shihusan and single herb on proliferation of retinal cells were observed with MTT test. The changes of mitochondrial membrane potential（MMP） of cultured retinal cells labeled with Rhodamine123 were detected by laser scanning confocal microscopy when they were treating with different herbs. Results The retinal cells adhered, spread and grew well in vitro. Shihusan groups（ 1.58 g·L^-1and 0.50 g·L^-1 ） and Dendrobium group（5.00 g·L^-1 ） could promote the activity of human retinal cells with 21.8% , 14. 2% and 13.4% , respectively. Moreover, Shihusan could raise MMP of retinal cells about 13.5% , and resist the decline of MMP induced by glutamate from 46.4% to 16.2%. Conclusions Shihusan can promote the activity of human retinal cells, and resist the damage of glutamate. There-fore,it may be a choice to treat retinal degenerated diseases.