中国实用眼科杂志
中國實用眼科雜誌
중국실용안과잡지
CHINESE JOURNAL OF PRACTICAL OPHTHALMOLOGY
2013年
8期
1072-1076
,共5页
郭小东%刘洁%胡文%王育君%杨虹瑜
郭小東%劉潔%鬍文%王育君%楊虹瑜
곽소동%류길%호문%왕육군%양홍유
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
人參皂甙Rg1%視網膜神經節細胞-5%穀氨痠毒性作用%蛋白激酶B
인삼조대Rg1%시망막신경절세포-5%곡안산독성작용%단백격매B
Ginsenoside Rg1%RGC-5 cell line%Glutamate excitotoxicity%Protein kinase B
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Rg1预处理后,MTT法观察RGCs的存活率,Annexin V/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Rg1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Rg1处理组比较存活率的差异有统计学意义(P ＜0.05); Rg1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)％；Rg1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); Rg1处理组Akt mRNA和蛋白表达水平均高于Glu和正常对照组.结论 人参皂苷Rg1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.
目的 探討人參皂苷Rgl對穀氨痠毒性損傷大鼠視網膜神經節細胞株RGC-5的保護作用及信號通路.方法 建立RGC-5穀氨痠興奮毒損傷模型,人參皂苷Rg1預處理後,MTT法觀察RGCs的存活率,Annexin V/PI染色流式細胞儀檢測細胞凋亡,RT-PCR檢測各組蛋白激酶B(Akt) mRNA錶達,免疫印跡法觀察Rg1預處理對Akt蛋白錶達的影響.結果 Glu組細胞吸光度值減少,細胞存活率為(41.9±5.4)％,與正常對照和Rg1處理組比較存活率的差異有統計學意義(P ＜0.05); Rg1處理組細胞的吸光度值比Glu組上升,細胞存活率提高到(68.2±4.7)％；Glu組壞死和凋亡晚期細胞率為(32.8±5.1)％,早期凋亡率為(29.4±2.1)％；Rg1組壞死和凋亡晚期細胞率為(9.2±1.1)％,早期凋亡率為(16.5±2.8)％,差異有統計學意義(P ＜0.05); Rg1處理組Akt mRNA和蛋白錶達水平均高于Glu和正常對照組.結論 人參皂苷Rg1可通過激活RGC-5細胞內Akt信號通路對抗穀氨痠引起興奮毒性作用,實現保護作用.
목적 탐토인삼조감Rgl대곡안산독성손상대서시망막신경절세포주RGC-5적보호작용급신호통로.방법 건립RGC-5곡안산흥강독손상모형,인삼조감Rg1예처리후,MTT법관찰RGCs적존활솔,Annexin V/PI염색류식세포의검측세포조망,RT-PCR검측각조단백격매B(Akt) mRNA표체,면역인적법관찰Rg1예처리대Akt단백표체적영향.결과 Glu조세포흡광도치감소,세포존활솔위(41.9±5.4)％,여정상대조화Rg1처리조비교존활솔적차이유통계학의의(P ＜0.05); Rg1처리조세포적흡광도치비Glu조상승,세포존활솔제고도(68.2±4.7)％；Glu조배사화조망만기세포솔위(32.8±5.1)％,조기조망솔위(29.4±2.1)％；Rg1조배사화조망만기세포솔위(9.2±1.1)％,조기조망솔위(16.5±2.8)％,차이유통계학의의(P ＜0.05); Rg1처리조Akt mRNA화단백표체수평균고우Glu화정상대조조.결론 인삼조감Rg1가통과격활RGC-5세포내Akt신호통로대항곡안산인기흥강독성작용,실현보호작용.
Objective To investigate the protective effect of ginsenoside Rgl antagonizes glutamate 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 analyzed using RT-PCR and immunoblotting.Results Cell absorbance value of Glu group was decreased,the cell survival rate was (41.9±5.4)％,there was statistically significant difference between 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 protein expression level of Akt in Rglgroup were higher than those of Glu and control group.Conclusion Rgl can protect RGC-5 against glutamate excitotoxicity effects through activation the Akt signaling pathway.
