氢离子对视网膜氧化应激损伤的保护作用机制研究

Protective effects of hydrogen ions on retina oxidative stress injury

目的探讨氢离子对氧化应激诱导的视网膜损伤的保护机制。方法 C57BL/6J雄性小鼠30只,随机分为对照组、模型组和治疗组,其中治疗组给予氢离子水灌胃7 d、模型组给予蒸馏水灌胃7 d后均建立视网膜损伤动物模型,对照组仅用蒸馏水灌胃,之后按前述方法分别持续灌胃5 d后处死,HE染色观察视网膜形态;二氢乙啶染色观察视网膜中活性氧的含量;剥离视网膜,Western blot检测氧化应激标志性蛋白ogg1、Sirt3及衰老相关蛋白P53、P21和P16的表达。结果视网膜HE染色结果显示,治疗组视网膜黑色玻璃膜疣沉积与模型组相比少且小;治疗组中氧化应激标志性蛋白ogg1相对表达量(0.63±0.04)明显低于模型组(0.96±0.02),差异具有统计学意义(P<0.05),与对照组(0.42±0.05)相比差异无统计学意义(P>0.05)。二氢乙啶染色结果表明,治疗组的视网膜的活性氧含量较模型组低;治疗组中Sirt3蛋白相对表达量(1.02±0.05)较模型组(0.55±0.04)显著升高,差异具有统计学意义(P<0.05);治疗组中衰老相关蛋白P53、P21、P16的相对表达量(0.53±0.05、0.40±0.01、0.71±0.01)较模型组(1.06±0.04、0.70±0.03、1.29±0.05)均显著降低,差异均具有统计学意义(均为P<0.05)。结论氢离子通过减少Sirt3表达的下调和抑制衰老对氧化应激诱导的视网膜损伤起保护作用。

Objective To investigate the protective mechanism of hydrogen ion on oxidative stress-induced retinal injulsn Methods Together 30 C57BL/SJ male mice were randomly divided into the control group, the model group and the treatment group. The treatment group was given the hydrogen ion water for 7 days and the model group was given the distilled water for 7 days, respectively, and both groups were estab- lished animal model of retinal injury, and the control group was only treated with distilled water. The mice were sacrificed after continuous gavaging 5 days according to the foregoing methods, and the retinal morphology was obserced by HE staining;DHE staining was used to observe the content of reactive oxygen species in the retina;the retina was collected, and Western blot was used to detect the expression of oxidative stress marker protein oggl, Sift3 and senescence related protein P53, P21 and P15. Results The retinal HE staining showed that the deposition of retinal black drusen in the treat- merit group was smaller than that in the model group;meanwhile,the relative expression of the oxidative stress marker protein oggl in the treatment group（0.53 _＋0.04） was sig- nificantly lower than that in the model group （0.95 ± 0.02）, and the difference was statis- tically significant（P 〈0.05）,but there was no significant difference （P 〉 0. 05）with con- trol group （0.42 ± 0.05 ） ; the result of DHE staining showed that the ROS content of the treatment group retina was lower than that of the model group. The relative expression of Sift3 in the treatment group （ 1.02± 0. 05 ） is significantly higher than that in the model group（0.55 ±0. 04 ）, and the difference was statistically significant （P 〈 0.05 ）, while the relative expression of senescence-associated proteins in the treatment group P53, P21 and P15 （0.53 ± 0.05,0.40 ± 0.01,0.71 ±0.01 ） were significantly lower than those in the mod- el group （ 1.05 ± 0.04,0.70 ±0.03,1.29 ±0.05）, and the difference was statistically significant（all P 〈 0. 05）. Conelusion Hydrogen ion can protect oxidative stress-induced retinal damage by decreasing down-regnlation of Sift3 and inhibiting senescence.