本文選題：急性缺氧 + 大鼠視網(wǎng)膜��； 參考：《青海大學(xué)》2017年碩士論文

【摘要】：目的:利用低壓艙模擬高原急性缺氧環(huán)境對(duì)大鼠視網(wǎng)膜的影響,探討白藜蘆醇對(duì)大鼠視網(wǎng)膜低氧損傷的保護(hù)作用及機(jī)制。方法:將72只SD大鼠隨機(jī)分成3組:常氧對(duì)照組(n=24)、低氧模型組(n=24)、低氧+白藜蘆干預(yù)組(n=24)。常氧對(duì)照組大鼠在常氧環(huán)境下正常喂養(yǎng)(西寧,海拔2260 m),低氧模型組及低氧+白藜蘆醇干預(yù)組大鼠置于低壓氧艙內(nèi)(模擬5000 m的海拔高度),24 h/d。低氧+白藜蘆醇干預(yù)組大鼠腹腔注射白藜蘆醇(30mg·kg-1,1次/d)。7 d后剝離完整視網(wǎng)膜并取3 mm視神經(jīng),光鏡下觀察大鼠視網(wǎng)膜組織形態(tài)變化,電鏡觀察視神經(jīng)超微結(jié)構(gòu)變化;酶聯(lián)免疫吸附測定法(ELISA)檢測大鼠視網(wǎng)膜組織中過氧化物歧化酶(Superoxide Dismutase,SOD)、谷胱甘肽過氧化物酶(Glutathione peroxidase,GSH-PX)的活性;免疫組織化學(xué)法觀察大鼠視網(wǎng)膜組織中膠質(zhì)纖維酸性蛋白(Glial fibrillary acidic protein,GFAP)、低氧誘導(dǎo)因子-1(Hypoxia-inducible factor-1,HIF-1)的表達(dá);RT-PCR方法檢測HIF、核轉(zhuǎn)錄因子-kappaB(Nuclear factor-kappa B1,NF-?B)、SOD與腦源性神經(jīng)營養(yǎng)因子(brain-derived neurotrophic factor,BDNF)mRNA的表達(dá)。結(jié)果:1、低氧模型組大鼠視網(wǎng)膜形態(tài),視神經(jīng)超微結(jié)構(gòu)較常氧對(duì)照組發(fā)生明顯變化;而低氧+白藜蘆醇干預(yù)組大鼠視網(wǎng)膜形態(tài)及視神經(jīng)受損明顯比低氧模型組減輕。2、與常氧對(duì)照組相比,低氧模型組大鼠視網(wǎng)膜組織中GSH-PX活性顯著下降(P=0.001),GFAP與HIF-1的表達(dá)增多,HIF、BDNF、NF-?B的mRNA表達(dá)量均顯著增加(P值分別是0.034,0.001,0.002),SOD活性及SOD mRNA的表達(dá)量無明顯差別(P值分別為0.088,0.262)。與低氧模型組對(duì)比,低氧+白藜蘆醇干預(yù)組大鼠視網(wǎng)膜GSH-PX活性顯著增強(qiáng)(P=0.004),GFAP與HIF-1的表達(dá)減少;HIF與BDNF的mRNA表達(dá)量顯著減少(P分別為0.001,0.021),差異有統(tǒng)計(jì)學(xué)意義,而SOD的活性及NF-?B、SOD mRNA表達(dá)量無明顯差異(P值分別為0.094,0.090,0.887)。結(jié)論:1)高原低壓缺氧環(huán)境對(duì)大鼠的視網(wǎng)膜功能有損傷作用,且其機(jī)制不是單一的,是多因素共同作用的結(jié)果。2)白藜蘆醇對(duì)高原急性缺氧誘導(dǎo)的視網(wǎng)膜損傷有干預(yù)作用,可能與其增強(qiáng)GSH-PX的酶活性,增強(qiáng)對(duì)自由基的清除以及調(diào)節(jié)HIF-1、BDNF與GFAP的表達(dá)有關(guān)。
[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 鹵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.
【學(xué)位授予單位】：青海大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R774

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