远志皂苷元对氧化应激损伤的视网膜神经节细胞的保护作用  

Protective effect of senegenin on retinal ganglion cells in oxidative stress induced injury

作　　者：别曼[1] 徐颖[2] 胡慧玲[3] 逯丹[1] 朱丽红[2] 戚仁斌[1] 王华东[1] 陆大祥[1] 

机构地区：[1]暨南大学医学院病理生理学系,暨南大学脑科学研究所,国家中药管理局三级科研实验室,广东广州510632 [2]暨南大学-香港大学脑功能与健康联合实验室,广东广州510632 [3]中山大学中山眼科中心,广东广州510060

出　　处：《中国病理生理杂志》2012年第6期1091-1096,共6页Chinese Journal of Pathophysiology

基　　金：国家重点基础研究发展计划(973计划)(No.2011CB707501)

摘　　要：目的:观察远志皂苷元(senegenin,Sen)对氧化应激损伤的视网膜神经节细胞(retinal ganglioncells,RGCs)的影响并初步探讨其作用机制。方法:采用上丘荧光金逆行标记RGCs后,体外原代RGCs混合细胞培养,随机分为control组、H2O2组、Sen+H2O2和Sen组。检测带荧光金荧光细胞的活力。同上述分组处理视网膜,用Hoechst 33258染色后观察视网膜细胞核形态的变化,Western blotting检测视网膜细胞cleaved caspase-3、细胞色素C及Bcl-2蛋白的表达。结果:与control组比较,Sen浓度在10、20和40μmol/L时,RGCs活力无明显变化(P>0.05),但Sen浓度达到80和160μmol/L时,RGCs活力明显下降,差异显著(P<0.01)。25、50、100和200μmol/L H2O2明显降低RGCs活力(P<0.05)。Sen浓度在10、20和40μmol/L时,对50μmol/L H2O2损伤的RGCs有较好的保护作用(P<0.05),其中40μmol/L Sen保护作用最为明显。Hoechst 33258染色表明Sen可以减少H2O2引起的视网膜细胞凋亡。Western blotting结果表明Sen促进Bcl-2蛋白的表达,降低线粒体细胞色素C的释放,下调cleaved caspase-3的表达。结论:Sen保护RGCs对抗氧化应激引起的损伤,其机制可能与其增强Bcl-2蛋白表达和减少氧化应激引起的细胞凋亡有关。AIM： To evaluate the effect of senegenin （Sen） on H2O2-treated retinal ganglion cells （RGCs） and to explore its underlying mechanisms. METHODS： RGCs were retrograde labeled by injection of fluorogold into the superior colliculi of SD rats on the postnatal day 3. On the postnatal days 6 to 8, the retinas were dissociated with papain and cultured. Primary RGCs cultured in vitro were treated with H2O2 and/or various doses of Sen. The viability of RGCs was evaluated by counting the fluorescence-labeled neurons under microscope. The morphological changes of the nuclei in the retinal neurons were observed by Hoechst 33258 staining. Western blotting was applied to determine the expression of cleaved caspase-3, cytochrome C and Bcl-2 in cultured retinal neurons. RESULTS： Compared with the control cells, Sen at doses of 10, 20 or 40 μmol/L had no toxicity to RGCs （P〉0.05）. However, Sen at doses of 80 and 160 μmol/L had significant toxicity to RGCs （P〈0.01）. Compared with H2O2-injured group, Sen at doses of 10, 20 and 40 μmol/L effectively protected against H2O2-induced injury in RGCs （P〈0.05） with the best efficiency at 40 μmol/L. Hoechst 33258 staining showed that the neuronal apoptosis caused by H2O2 was reduced by Sen. The results of Western blotting showed an up-regulation of Bcl-2, and decreased cytochrome C and cleaved caspase-3 levels by Sen in H2O2-treated retinal neurons. CONCLUSION： Sen is able to protect RGCs from H2O2-induced injury by enhancing Bcl-2 expression and inhibiting cell apoptosis.

关 键 词：视网膜神经节细胞 远志皂苷元 氧化性应激 

分 类 号：R363[医药卫生—病理学]