Effects of One Berry Herbal Beverage on Retinal Cells and Antioxidant Capacity in Optic Nerve Crushed Mice
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摘要: 为探究一款浆果草本饮料的护眼功效，采用视神经挤压伤模型小鼠，术前术后分别灌胃7 d，实验组灌胃饮料剂量为0.1、1、10、100 mg/kg，对照组灌胃磷酸盐缓冲溶液，利用视网膜平铺免疫荧光染色、视网膜蛋白免疫印迹检测和活性氧（ROS）荧光探针检测方法，测定灌胃14 d后小鼠视网膜节细胞的存活数目、视网膜蛋白表达情况和视网膜氧化应激水平。结果表明，该护眼功能饮料对视网膜神经节细胞（RGCs）存活量的影响是浓度依赖性的，4个剂量组中只有10 mg/kg剂量组RGCs存活量与对照组相比显著增加（P<0.05），提高了约8.25%；在此剂量下，视网膜的免疫炎症细胞小胶质细胞的激活和极化无明显变化；但视网膜内抗氧化应激相关蛋白锰超氧歧化酶（MnSOD）、血红素氧合酶1（HO-1）和核因子样蛋白（Nrf-2）有明显提升；同时实验组挤压伤眼与自身对照对侧眼的视网膜的ROS水平分别降低约25.62%与16.95%。表明此护眼功能饮料可通过保持视网膜神经节细胞活性，增强视网膜的抗氧化能力及降低视网膜活性氧水平从而达到护目功效。Abstract: To explore the eye protection effect of one berry herbal beverage,optic nerve crush model mice were used. The beverage was administered intragastrically daily for 7 days before and after optic nerve crushed,respectively. The gavage dose of the experimental group was 0.1,1,10,100 mg/kg,and the control group was gavage with phosphate buffer solution. Immunofluorescence staining of retinal stretching preparation,western blot analysis and reactive oxygen species(ROS)fluorescent probe were applied to determine the survival of retinal ganglion cells,retinal protein expression and retinal oxidative stress level of mice after 14 days’ intragastric administration. The results showed that this eye-protecting beverage affected retinal ganglion cells(RGCs)in a dose-dependent manner. Among the 4 dose groups,only 10 mg/kg group significantly increased the survival of retinal ganglion cells(RGCs)by about 8.25% compared with the control group(P<0.05). Under the above-mentioned dosage,there was no significant change observed in the activation and polarization of microglia,a retinal immunoinflammatory cell. But the content of some oxidative stress related proteins in retina,including manganese superoxide dismutase(MnSOD),heme oxygenase 1(HO-1)and nuclear factor erythroid 2-related factor 2(Nrf-2),increased significantly. Meanwhile,the ROS level of retina of experimental group was decreased by about 25.62% and 16.95% for crush injured eye and self-control contralateral eye respectively. Therefore,this beverage could protect eyes by maintaining the activity of retinal ganglion cells,enhancing the antioxidant capacity of retina and reducing the level of reactive oxygen species of retina.
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