人参皂甙Rgl激活RGC-5细胞Akt信号通路拮抗谷氨酸兴奋毒性的实验研究  

Experimental study of ginsenoside Rgl antagonizes glutamate excitotoxicity on RGC-5 cells by Akt signaling pathway activation

作　　者：郭小东 刘洁[2] 胡文 王育君 杨虹瑜 

机构地区：[1]广东省揭阳市慈云医院,广东522031 [2]中国医科大学实验技术中心,沈阳110001

出　　处：《中国实用眼科杂志》2013年第8期1072-1076,共5页Chinese Journal of Practical Ophthalmology

基　　金：广东省医学科研基金人参皂甙Rgl治疗视神经损伤性疾病实验研究（A2011103）

摘　　要：目的探讨人参皂苷Rgl对谷氨酸毒性损伤大鼠视网膜神经节细胞株RGC-5的保护作用及信号通路。方法建立RGC-5谷氨酸兴奋毒损伤模型，人参皂苷Rgl预处理后，MTF法观察RGCs的存活率，AnnexinV／PI染色流式细胞仪检测细胞凋亡，RT—PCR检测各组蛋白激酶B（Akt）mRNA表达，免疫印迹法观察Rgl预处理对Akt蛋白表达的影响。结果Glu组细胞吸光度值减少，细胞存活率为（41．9±5．4）％，与正常对照和Rgl处理组比较存活率的差异有统计学意义（P〈0．05）；Rgl处理组细胞的吸光度值比Glu组上升，细胞存活率提高到（68．2±4．7）％；Glu组坏死和凋亡晚期细胞率为（32．8±5．1）％，早期凋亡率为（29．4±2．1）％；Rgl组坏死和凋亡晚期细胞率为（9．2±1．1）％，早期凋亡率为（16．5±2．8）％，差异有统计学意义（P〈0．05）；Rgl处理组AktmRNA和蛋白表达水平均高于Glu和正常对照组。结论人参皂苷Rgl可通过激活RGC-5细胞内Akt信号通路对抗谷氨酸引起兴奋毒性作用，实现保护作用。Objective To investigate the protective effect of ginsenoside Rgl antagonizes gluta- mate excitotoxicity on rat retinal ganglion cell line RGC-5 and related signal pathway. Methods RGC-5 glutamate excitotoxicity model was built and pretreated with ginsenoside Rgl.The survival rate of RGCs was examined by MTT method. Cell apoptosis was tested by flow cytometry with Annexin V/PI staining kit. The expression of protein kinase B （Akt） mRNA and protein were ana- lyzed using RT-PCR and immunoblotting. Results Cell absorbance value of Glu group was de- creased, the cell survival rate was （41.9±5.4）%, there was statistically significant difference be- tween normal control group and Rgl treatment group in survival. （P〈0.05）. Cell absorbance value in Rgl group was higher than that of the Glu group, the cell survival rate was increased to （68.2± 4.7）%. Necrosis and late apoptosis rate was （32.8± 5.1）% ,early apoptosis rate was （29.4±2.1）% in Glu group; Necrosis and late apoptosis rate was （9.2 ± 1.1）%, early apoptosis rate was （16.5±2.8）% in Rgl group, there was significant difference between them （P〈0.05）; The mRNA and pro- tein expression level of Akt in Rglgroup were higher than those of Glu and control group. Conclu- sion Rgl can protect RGC-5 against glutamate excitotoxicity effects through activation the Akt signal- ing pathway.

关 键 词：人参皂甙RG1 视网膜神经节细胞-5 谷氨酸毒性作用 蛋白激酶B 

分 类 号：R743.3[医药卫生—神经病学与精神病学]