本文選題：糖尿病視網(wǎng)膜病變 + 缺血再灌注損傷��； 參考：《武漢大學》2013年博士論文

【摘要】：視網(wǎng)膜病變是一類危害人類生活質(zhì)量的較普遍的疾病。視網(wǎng)膜缺血再灌注損傷在糖尿病視網(wǎng)膜病變、青光眼等多種視網(wǎng)膜病變中均有發(fā)生。視網(wǎng)膜缺血再灌注會導致視網(wǎng)膜神經(jīng)退化、血管退化和膠質(zhì)細胞激活等病理現(xiàn)象,但是其分子機制目前尚未完全清楚,并且還缺乏有效的治療藥物。 炎癥反應是缺血再灌注導致視網(wǎng)膜損傷的重要機制之一。同時,慢性炎癥反應對糖尿病視網(wǎng)膜病變的發(fā)生發(fā)展具有促進作用。組蛋白修飾能夠調(diào)節(jié)基因表達。因此猜想炎癥因子在病變視網(wǎng)膜中的過量和持續(xù)表達可能與組蛋白修飾相關。 本論文采用三種實驗模型,包括高眼壓誘導的視網(wǎng)膜缺血再灌注模型、STZ(鏈脲佐菌素)誘導的糖尿病模型和高糖刺激的米勒膠質(zhì)細胞模型來研究視網(wǎng)膜損傷的炎癥發(fā)生機制。主要通過PASH和免疫染色等方法檢測視網(wǎng)膜病理變化,應用蛋白質(zhì)印跡、聚合酶鏈式反應和染色質(zhì)免疫共沉淀等技術研究炎癥反應、組蛋白修飾及組蛋白異構體在視網(wǎng)膜損傷以及體外細胞模型中的變化。姜黃素和米林霉素兩種藥物都具有抗炎作用,本論文采用這兩種藥物對視網(wǎng)膜損傷的動物模型和細胞模型進行預防和干預治療,研究這些藥物的作用靶標以及組蛋白修飾和炎癥反應在視網(wǎng)膜損傷中的相互聯(lián)系。 本論文發(fā)現(xiàn),在損傷前口服0.01%、0.05%或者0.25%的姜黃素能夠顯著的抑制視網(wǎng)膜缺血再灌注損傷引起的神經(jīng)節(jié)細胞層的細胞缺失；損傷前或者損傷后口服0.05%的姜黃素都能夠抑制損傷引起的血管退化；損傷前口服0.05%的姜黃素抑制了缺血再灌注損傷引起的視網(wǎng)膜神經(jīng)節(jié)細胞層的細胞凋亡、β-tubulin Ⅲ下調(diào)、NF-κB和STAT3(?)言號通路激活以及炎癥因子MCP-1表達量上調(diào),但是對損傷引起的米勒細胞激活和ERK信號通路激活沒有顯著作用。因此,姜黃素對缺血再灌注損傷導致的視網(wǎng)膜神經(jīng)和血管退化的保護作用是通過抑制炎癥信號通路的激活以及減少炎癥因子的過量表達實現(xiàn)的。 本論文還發(fā)現(xiàn)早期病史的糖尿病大鼠視網(wǎng)膜內(nèi)組蛋白的乙�；揎椝斤@著升高,并伴隨組蛋白乙酰化酶表達量上調(diào)和組蛋白2類去乙�；副磉_量下調(diào)。并且證明糖尿病大鼠視網(wǎng)膜內(nèi)高水平的組蛋白H3K9和H3K18乙�；糠侄ㄎ辉诿桌占毎稀Ｔ隗w外高糖培養(yǎng)的米勒細胞系rMC-1中也發(fā)現(xiàn)組蛋白乙�；叩默F(xiàn)象,同時GFAP、p-STAT3(Tyr)和NF-KB-p65的蛋白水平以及炎癥因子TNFα和MCP-1的mRNA水平也顯著上調(diào)。另外還發(fā)現(xiàn)米林霉素通過抑制高糖導致的GFAP、TNFα和MCP-1啟動子上組蛋白H3K18的乙�；�,抑制了這三個基因的轉(zhuǎn)錄。這些結果表明高糖導致米勒細胞中炎癥因子啟動子上的組蛋白乙�；缴�,因而使米勒細胞產(chǎn)生炎癥反應；同時米林霉素對糖尿病視網(wǎng)膜病變的保護作用也來源于對組蛋白乙�；降囊种啤�
[Abstract]:Retinopathy is a common disease that endangers the quality of human life. Retinal ischemia reperfusion injury occurs in many kinds of retinopathy such as diabetic retinopathy, glaucoma and so on. Retinal ischemia-reperfusion can lead to retinal nerve degeneration, vascular degeneration and glial cell activation, but the molecular mechanism of retinal ischemia reperfusion is not fully understood, and there is a lack of effective treatment drugs. Inflammation is one of the important mechanisms of retinal injury induced by ischemia reperfusion. At the same time, chronic inflammatory reaction can promote the development of diabetic retinopathy. Histone modification can regulate gene expression. It is assumed that excessive and persistent expression of inflammatory factors in the pathological retina may be related to histone modification. In this paper, three experimental models were used to study the inflammatory mechanism of retinal injury, including the diabetic model induced by high intraocular pressure (IOP) induced retinal ischemia and reperfusion (STZ) and the Hans Muller glial cell model stimulated by high glucose. The pathological changes of retina were detected by PASH and immunostaining. The inflammatory reaction was studied by Western blotting, polymerase chain reaction and chromatin immunoprecipitation. Histone modification and histone isomer changes in retinal injury and in vitro cell model. Curcumin and milinomycin both have anti-inflammatory effects. In this paper, the two drugs were used to prevent and intervene the animal model and cell model of retinal injury. To study the effects of these drugs and the correlation between histone modification and inflammation in retinal injury. In this study, we found that oral administration of 0.05% or 0.25% curcumin before injury could significantly inhibit the loss of neurons in the ganglion cell layer induced by retinal ischemia-reperfusion injury. Before and after injury, 0.05% of curcumin could inhibit the vascular degeneration induced by injury, and before injury, 0.05% of curcumin inhibited the apoptosis of retinal ganglion cell layer induced by ischemia-reperfusion injury, and 尾 -tubulin 鈪，

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