本文選題：AMPK + 蛋白激酶通路�。� 參考：《大連醫(yī)科大學(xué)》2011年碩士論文

【摘要】：目的:單磷酸腺苷激活的蛋白激酶(AMPK)通路通過(guò)抑制能量消耗通路和激活能量再生通路,在能源動(dòng)態(tài)平衡調(diào)控中起著中樞性調(diào)節(jié)作用。AMPK通路在生理刺激(如運(yùn)動(dòng))、激素(如脂聯(lián)素,瘦素等)、以及病理?xiàng)l件(如缺糖,缺氧,氧化)等情況下可以被激活。作為脂類和血糖代謝調(diào)節(jié)作用的樞紐,AMPK同時(shí)被認(rèn)為是治療肥胖、Ⅱ型糖尿病和癌癥的關(guān)鍵靶點(diǎn)。一些研究表明AMPK具有神經(jīng)保護(hù)作用,到目前為止,還沒(méi)有關(guān)于AMPK通路對(duì)視網(wǎng)膜病變的保護(hù)作用方面的研究報(bào)道。氧化應(yīng)激損傷是多種視網(wǎng)膜病變病理變化的誘因。減少和抑制氧化應(yīng)激和其所誘發(fā)的神經(jīng)元凋亡是目前很多對(duì)視網(wǎng)膜保護(hù)措施的機(jī)理。急性光損傷目前是公認(rèn)的模擬氧化應(yīng)激引起的視神經(jīng)元凋亡的動(dòng)物模型。本論文的目的旨在研究AMPK通路對(duì)視網(wǎng)膜神經(jīng)元在氧化應(yīng)激條件下的保護(hù)作用和機(jī)制。 方法:實(shí)驗(yàn)用小白鼠以玻璃體內(nèi)注射或皮下注射AMPK激動(dòng)劑二甲雙胍和PBS對(duì)照,連續(xù)7天。在急性光損傷模型中,小白鼠被置于4000 lux強(qiáng)光下照射4小時(shí)(從8:00pm到12:00am)。在給藥或急性光損傷5天后,用視網(wǎng)膜電圖(ERG)法記錄視網(wǎng)膜感光細(xì)胞的功能。組織學(xué)研究用于觀察給藥和或光損傷后視神經(jīng)元的凋亡情況和組織結(jié)構(gòu)。蛋白印記法和免疫熒光法用于檢測(cè)小白鼠視網(wǎng)膜內(nèi)AMPK及其下游通路的激活和表達(dá)水平。應(yīng)用聚合酶鏈?zhǔn)椒磻?yīng)(PCR)測(cè)量線粒體DNA拷貝量。 結(jié)果:玻璃體和皮下注射二甲雙胍可以激活視網(wǎng)膜內(nèi)AMPK通路,并在急性光損傷模型中保護(hù)視網(wǎng)膜感光細(xì)胞的結(jié)構(gòu)和功能。進(jìn)一步的研究表明,激活A(yù)MP蛋白激酶通路誘導(dǎo)的細(xì)胞核和線粒體編碼基因的表達(dá)水平增加可能是其保護(hù)作用的機(jī)制。 結(jié)論:通過(guò)實(shí)驗(yàn)我們證明皮下注射二甲雙胍可以激活視網(wǎng)膜內(nèi)的AMPK蛋白激酶通路,說(shuō)明二甲雙胍可以通過(guò)血-視網(wǎng)膜屏障。而且,我們的研究發(fā)現(xiàn),激活A(yù)MPK通路可以強(qiáng)烈保護(hù)視神經(jīng)元,避免氧化應(yīng)激損傷誘導(dǎo)的神經(jīng)元凋亡。這些實(shí)驗(yàn)結(jié)果表明激活A(yù)MPK蛋白激酶通路,增加線粒體損傷修復(fù)或生物合成,可能成為治療或延遲視網(wǎng)膜退行性變的新方法。
[Abstract]:Aim: adenosine monophosphate activated protein kinase (AMPK) pathway inhibits energy consumption and activates energy regeneration pathway. AMPK pathway plays a central role in the regulation of energy homeostasis. AMPK pathway can be activated under physiological stimulation (such as motor stimulation, hormones (such as adiponectin, leptin, etc.), and pathological conditions (such as glucose deficiency, hypoxia, oxidation). AMPK, which acts as a key regulator of lipid and glucose metabolism, is also considered a key target for the treatment of obesity, type 2 diabetes and cancer. Some studies have shown that AMPK has neuroprotective effect, so far, there have been no reports on the protective effect of AMPK pathway on retinopathy. Oxidative stress injury is the inducement of pathological changes of various retinopathy. Reduction and inhibition of oxidative stress and neuronal apoptosis induced by oxidative stress are the mechanisms of many protective measures for retina. Acute light injury is recognized as an animal model of apoptosis of optic neurons induced by oxidative stress. The aim of this study was to investigate the protective effect and mechanism of AMPK pathway on retinal neurons under oxidative stress. Methods: rats were treated with intravitreous injection or subcutaneous injection of AMPK agonist metformin and PBS for 7 days. In the acute light injury model, the mice were exposed to 4000 lux intense light for 4 hours (from 8:00pm to 12: 00 amps). The function of retinal photoreceptor cells was recorded by electroretinogram (ERG) 5 days after administration or acute light injury. Histological study was used to observe the apoptosis and structure of optic neurons after drug administration or light injury. Protein imprinting and immunofluorescence were used to detect the activation and expression of AMPK and its downstream pathway in rat retina. Mitochondrial DNA copies were measured by polymerase chain reaction (PCR). Results: vitreous and subcutaneous injection of metformin could activate the AMPK pathway in the retina and protect the structure and function of the photoreceptor cells in the acute light injury model. Further studies suggest that the increased expression of nuclear and mitochondrial coding genes induced by activation of AMP protein kinase pathway may be the mechanism of its protective effect. Conclusion: subcutaneous injection of metformin can activate the AMPK protein kinase pathway in the retina, suggesting that metformin can pass the blood-retinal barrier. Furthermore, we found that activation of AMPK pathway can strongly protect optic neurons from apoptosis induced by oxidative stress. These results suggest that activating the AMPK protein kinase pathway and increasing mitochondrial damage repair or biosynthesis may be a new method to treat or delay retinal degeneration.
【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R779.1

【共引文獻(xiàn)】

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本文編號(hào)：1875921