急进高原大鼠视网膜功能变化及白藜芦醇干预效应研究

发布时间：2018-05-29 22:23

本文选题：急性缺氧 + 大鼠视网膜　；参考：《青海大学》2017年硕士论文

【摘要】：目的:利用低压舱模拟高原急性缺氧环境对大鼠视网膜的影响,探讨白藜芦醇对大鼠视网膜低氧损伤的保护作用及机制。方法:将72只SD大鼠随机分成3组:常氧对照组(n=24)、低氧模型组(n=24)、低氧+白藜芦干预组(n=24)。常氧对照组大鼠在常氧环境下正常喂养(西宁,海拔2260 m),低氧模型组及低氧+白藜芦醇干预组大鼠置于低压氧舱内(模拟5000 m的海拔高度),24 h/d。低氧+白藜芦醇干预组大鼠腹腔注射白藜芦醇(30mg·kg-1,1次/d)。7 d后剥离完整视网膜并取3 mm视神经,光镜下观察大鼠视网膜组织形态变化,电镜观察视神经超微结构变化;酶联免疫吸附测定法(ELISA)检测大鼠视网膜组织中过氧化物歧化酶(Superoxide Dismutase,SOD)、谷胱甘肽过氧化物酶(Glutathione peroxidase,GSH-PX)的活性;免疫组织化学法观察大鼠视网膜组织中胶质纤维酸性蛋白(Glial fibrillary acidic protein,GFAP)、低氧诱导因子-1(Hypoxia-inducible factor-1,HIF-1)的表达;RT-PCR方法检测HIF、核转录因子-kappaB(Nuclear factor-kappa B1,NF-?B)、SOD与脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)mRNA的表达。结果:1、低氧模型组大鼠视网膜形态,视神经超微结构较常氧对照组发生明显变化;而低氧+白藜芦醇干预组大鼠视网膜形态及视神经受损明显比低氧模型组减轻。2、与常氧对照组相比,低氧模型组大鼠视网膜组织中GSH-PX活性显著下降(P=0.001),GFAP与HIF-1的表达增多,HIF、BDNF、NF-?B的mRNA表达量均显著增加(P值分别是0.034,0.001,0.002),SOD活性及SOD mRNA的表达量无明显差别(P值分别为0.088,0.262)。与低氧模型组对比,低氧+白藜芦醇干预组大鼠视网膜GSH-PX活性显著增强(P=0.004),GFAP与HIF-1的表达减少;HIF与BDNF的mRNA表达量显著减少(P分别为0.001,0.021),差异有统计学意义,而SOD的活性及NF-?B、SOD mRNA表达量无明显差异(P值分别为0.094,0.090,0.887)。结论:1)高原低压缺氧环境对大鼠的视网膜功能有损伤作用,且其机制不是单一的,是多因素共同作用的结果。2)白藜芦醇对高原急性缺氧诱导的视网膜损伤有干预作用,可能与其增强GSH-PX的酶活性,增强对自由基的清除以及调节HIF-1、BDNF与GFAP的表达有关。
[Abstract]:Aim: to investigate the protective effect and mechanism of resveratrol on retinal hypoxic injury in rats by simulating the acute hypoxia environment at high altitude. Methods: 72 Sprague-Dawley rats were randomly divided into 3 groups: normoxic control group (n = 24), hypoxic model group (n = 24), hypoxia intervention group (n = 24). Rats in normoxic control group were fed in normoxic environment (Xining, altitude 2260 m), hypoxia model group and hypoxia resveratrol intervention group were placed in hypobaric oxygen chamber (simulated altitude of 5000 m) for 24 h / d. In the hypoxia resveratrol intervention group, 30 mg resveratrol was injected intraperitoneally to the rats. The intact retina was removed and the optic nerve was removed 3 mm after once a day. The morphological changes of the retina and the ultrastructure of the optic nerve were observed under light microscope and electron microscope. The activity of superoxide dismutase (SOD), glutathione peroxidase (Glutathione peroxidase) GSH-PX in the retina of rats was detected by enzyme-linked immunosorbent assay (Elisa). The expression of glial fibrillary acidic protein (Glial fibrillary acidic protein) and hypoxia-inducible factor-1 (HIF-1) in rat retina were observed by immunohistochemical method. RT-PCR was used to detect the expression of HIF-1, nuclear transcription factor -kappa B1, NF-BSOD and brain-derived neurotrophic factor (brain-derived neurotrophic factor) in rat retina. Results the retinal morphology and the ultrastructure of optic nerve in the hypoxia model group were significantly different from those in the normoxic control group. However, the retinal morphology and optic nerve damage in the hypoxic resveratrol group were significantly less than those in the hypoxic model group, which was significantly lower than that in the normoxic control group. In the hypoxic model group, the activity of GSH-PX in the retina decreased significantly. The expression of GSH-PX and HIF-1 in the retina of the hypoxic model group were significantly increased. The mRNA expression of NF-B was increased significantly (P = 0.034, 0.001, 0.002) and the expression of SOD mRNA was not significantly different (P = 0.088 0. 262, respectively). Compared with the hypoxic model group, the GSH-PX activity in the retina of the rats treated with resveratrol significantly increased the expression of GFAP and HIF-1 in the rat retina. The expression of mRNA in HIF and BDNF decreased significantly (P = 0.001 卤0.021), and the difference was statistically significant. However, there was no significant difference in the activity of SOD and the expression of mRNA in NF-mb (P = 0.094, 0.090, P = 0.887, respectively). Conclusion: (1) the hypobaric hypoxia at high altitude can injure the retinal function of rats, and the mechanism is not single, which is the result of multiple factors. 2) resveratrol can interfere with the retinal damage induced by acute hypoxia at high altitude. It may be related to the enhancement of GSH-PX activity, the enhancement of free radical scavenging and the regulation of HIF-1BDNF and GFAP expression.
【学位授予单位】：青海大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R774

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