石斛碱对过氧化氢诱导的RGC-5细胞氧化损伤的保护作用  

Effect of dendrobine on oxidative stress of RGC-5 induced by hydrogen peroxide

作　　者：毛黔华 许凤青 岳芳 张玮 石京山 Mao Qianhua;Xu Fengqing;Yue Fang;Zhang Wei;Shi Jingshan(Key Laboratory of Basic Pharmacology of Ministry of Education and Joint International Research Laboratory of Ethnomedicine of Ministry of Education,Zunyi Medical University,Zunyi Guizhou 563006,China;School of Pharmacy,Zunyi Medical University,Zunyi Guizhou 563006,China;China Pharmacology Society-Guizhou Pharmacology Union Laboratory,Zunyi Medical University,Zunyi Guizhou 563006,China)

机构地区：[1]遵义医科大学基础药理教育部重点实验室暨特色民族药教育部国际合作联合实验室,贵州遵义563006 [2]遵义医科大学药学院,贵州遵义563006 [3]遵义医科大学中国药理学会-贵州省药理学联合实验室,贵州遵义563006

出　　处：《遵义医科大学学报》2024年第8期768-774,782,共8页Journal of Zunyi Medical University

基　　金：国家自然科学基金资助项目(NO:81773739,U1812403)。

摘　　要：目的研究石斛碱对过氧化氢(H_(2)O_(2))诱导的视神经节细胞氧化损伤的保护作用。方法将培养的RGC-5细胞分为正常对照组、H_(2)O_(2)模型组、石斛碱1 nmol/L组、石斛碱10 nmol/L组及石斛碱100 nmol/L组。采用细胞技术试剂盒-8(CCK-8)检测RGC-5细胞活力;试剂盒法检测细胞中氧化和抗氧化物质的含量、细胞中ROS水平;蛋白免疫印迹法(Western Blot)检测细胞中核因子E2相关因子2(Nrf2)以及下游靶蛋白表达。结果石斛碱在1、10、100 nmol/L浓度对RGC-5细胞活力无毒性作用。H_(2)O_(2)从200μmol/L浓度开始显著降低RGC-5细胞活力。100 nmol/L石斛碱可显著降低H_(2)O_(2)诱导的RGC-5细胞中LDH外漏率、活性氧和丙二醛含量,显著升高超氧化物歧化酶和谷胱甘肽含量。石斛碱可诱导RGC-5细胞中Nrf2从胞浆向胞核转移,同时正向调控其下游抗氧化蛋白GPx1、GCLC、NQO 1和HO-1的蛋白表达。结论石斛碱对H_(2)O_(2)诱导的RGC-5细胞氧化损伤有明显的保护作用,其机制可能是石斛碱促进Nrf2核转位,正向调控Nrf2下游抗氧化蛋白表达发挥抗氧化作用。Objective To study the protective effect of dendrobine on H_(2)O_(2)-induced oxidative damage of optic ganglion cells.Methods The cultured RGC-5 cells were divided into normal control group,H_(2)O_(2)model group,dendrobine 1 nmol/L group,dendrobine 10 nmol/L group and dendrobine 100 nmol/L group,and the cell viability of RGC-5 cells was detected by CCK-8.The content of oxidation and antioxidant substances and the level of ROS in cells were detected by kit method.Western Blot was used to detect the expression of nuclear factor E2-related factor 2(Nrf2)and the downstream target protein.Results Dendrobine at the concentrations of 1,10 and 100 nmol/L has no toxic effect on RGC-5 cell viability.H_(2)O_(2)significantly reduced the activity of RGC-5 cells from the concentration of 200μmol/L.Dendrobine at the concentrations of 100 nmol/L can significantly reduce the leakage rate of LDH,the contents of reactive oxygen species and malondialdehyde in RGC-5 cells induced by H_(2)O_(2),and significantly increase the contents of superoxide dismutase and glutathione.Dendrobine can induce the transfer of Nrf2 from cytoplasm to nucleus in RGC-5 cells,and at the same time positively regulate the protein expression of its downstream antioxidant proteins GPx1,GCLC,NQO 1 and HO-1.Conclusion Dendrobine has obvious protective effect on H_(2)O_(2)-induced oxidative damage of RGC-5 cells.Its mechanism may be that Dendrobine promotes nuclear translocation of Nrf2,positively regulates the expression of downstream antioxidant protein on Nrf2 pathway,and thus plays an antioxidant role.

关 键 词：石斛碱 视神经节细胞 过氧化氢 氧化损伤 

分 类 号：R961[医药卫生—药理学]