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(xué)》2013年博士論文

【摘要】：視網(wǎng)膜病變是一類(lèi)危害人類(lèi)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(huì)導(dǎo)致視網(wǎng)膜神經(jīng)退化、血管退化和膠質(zhì)細(xì)胞激活等病理現(xiàn)象,但是其分子機(jī)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(yīng)是缺血再灌注導(dǎo)致視網(wǎng)膜損傷的重要機(jī)制之一。同時(shí),慢性炎癥反應(yīng)對(duì)糖尿病視網(wǎng)膜病變的發(fā)生發(fā)展具有促進(jìn)作用。組蛋白修飾能夠調(diào)節(jié)基因表達(dá)。因此猜想炎癥因子在病變視網(wǎng)膜中的過(guò)量和持續(xù)表達(dá)可能與組蛋白修飾相關(guān)。 本論文采用三種實(shí)驗(yàn)?zāi)Ｐ?包括高眼壓誘導(dǎo)的視網(wǎng)膜缺血再灌注模型、STZ(鏈脲佐菌素)誘導(dǎo)的糖尿病模型和高糖刺激的米勒膠質(zhì)細(xì)胞模型來(lái)研究視網(wǎng)膜損傷的炎癥發(fā)生機(jī)制。主要通過(guò)PASH和免疫染色等方法檢測(cè)視網(wǎng)膜病理變化,應(yīng)用蛋白質(zhì)印跡、聚合酶鏈?zhǔn)椒磻?yīng)和染色質(zhì)免疫共沉淀等技術(shù)研究炎癥反應(yīng)、組蛋白修飾及組蛋白異構(gòu)體在視網(wǎng)膜損傷以及體外細(xì)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(duì)視網(wǎng)膜損傷的動(dòng)物模型和細(xì)胞模型進(jìn)行預(yù)防和干預(yù)治療,研究這些藥物的作用靶標(biāo)以及組蛋白修飾和炎癥反應(yīng)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(xì)胞層的細(xì)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(xì)胞層的細(xì)胞凋亡、β-tubulin Ⅲ下調(diào)、NF-κB和STAT3(?)言號(hào)通路激活以及炎癥因子MCP-1表達(dá)量上調(diào),但是對(duì)損傷引起的米勒細(xì)胞激活和ERK信號(hào)通路激活沒(méi)有顯著作用。因此,姜黃素對(duì)缺血再灌注損傷導(dǎo)致的視網(wǎng)膜神經(jīng)和血管退化的保護(hù)作用是通過(guò)抑制炎癥信號(hào)通路的激活以及減少炎癥因子的過(guò)量表達(dá)實(shí)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�；副磉_(dá)量上調(diào)和組蛋白2類(lèi)去乙�；副磉_(dá)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乙酰化部分定位在米勒細(xì)胞上。在體外高糖培養(yǎng)的米勒細(xì)胞系rMC-1中也發(fā)現(xiàn)組蛋白乙�；叩默F(xiàn)象,同時(shí)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(guò)抑制高糖導(dǎo)致的GFAP、TNFα和MCP-1啟動(dòng)子上組蛋白H3K18的乙酰化水平,抑制了這三個(gè)基因的轉(zhuǎn)錄。這些結(jié)果表明高糖導(dǎo)致米勒細(xì)胞中炎癥因子啟動(dòng)子上的組蛋白乙�；缴�,因而使米勒細(xì)胞產(chǎn)生炎癥反應(yīng)；同時(shí)米林霉素對(duì)糖尿病視網(wǎng)膜病變的保護(hù)作用也來(lái)源于對(duì)組蛋白乙酰化水平的抑制。
[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，

本文編號(hào)：1846967