本文選題：載脂蛋白J + ox-LDL　； 參考：《河北北方學(xué)院》2017年碩士論文

【摘要】：現(xiàn)如今心血管疾病頻發(fā),已成為制約人們健康的一大難題。心血管疾病類型眾多,高頻發(fā)的疾病包括缺血性心肌病、高血壓以及動脈粥樣硬化(atherosclerosis,AS)等。通過調(diào)查,多數(shù)的心血管疾病和AS之間均有密切的關(guān)聯(lián)。國內(nèi)外業(yè)界人士從未停止過對冠心病的研究,據(jù)最新調(diào)研結(jié)果顯示,多數(shù)不必要的活性氧自由基(reactive oxygen species,ROS)是種種冠心病產(chǎn)生因素都可以激發(fā)的,活性氧進(jìn)而會產(chǎn)生誘發(fā)AS的病理過程。氧化低密度脂蛋白(ipoproteidensity low oxidizednl,ox-LDL)的功能多樣,不但可損傷血管內(nèi)皮、誘發(fā)細(xì)胞產(chǎn)生細(xì)胞因子,而且會同時(shí)造成細(xì)胞凋亡。載脂蛋白J(Apolipoprotein-J,ApoJ)是多存于人體組織與體液中的新型多功能分泌型的糖蛋白質(zhì)。它既可參與精子成熟、組織重構(gòu)、和補(bǔ)體活化的調(diào)控等生理病理過程,又可在細(xì)胞凋亡和脂質(zhì)運(yùn)輸中發(fā)揮作用。載脂蛋白J在抑制凋亡細(xì)胞方面有顯著作用,因此可保護(hù)心臟受損時(shí)的細(xì)胞。但是值得注意的是,即使載脂蛋白J與ox-LDL二者之間存在的聯(lián)系對于心肌細(xì)胞的生長與死亡起著決定性的作用,仍未完全引起足夠的重視。目的:闡明ox-LDL對心肌細(xì)胞的影響及細(xì)胞內(nèi)信號轉(zhuǎn)導(dǎo)機(jī)制,并探討高表達(dá)ApoJ是否可降低ox-LDL對新生大鼠心室肌細(xì)胞(neonatal rat ventricular cells,NRVCs)的影響。方法:利用帶有ApoJ基因的腺病毒感染NRVCs使ApoJ高表達(dá)。用CaMKⅡ抑制劑KN93和SOD類似物Mn(ⅡI)TBAP進(jìn)行前期預(yù)處理,劃分心肌細(xì)胞為:正常對照組、ox-LDL組、ox-LDL聯(lián)合ApoJ組、ApoJ組、ox-LDL聯(lián)合KN93組、ox-LDL聯(lián)合Mn(ⅡI)TBAP組、H2O2組、H2O2+ApoJ組。通過MTT檢測法和Caspase-3/7活性檢測試劑盒檢測心肌細(xì)胞活性和凋亡變化;利用Western-blot檢測心肌細(xì)胞Nox2/gp91phox、P47和CaMKⅡ表達(dá)情況,利用CaMKⅡ活性檢測試劑盒檢測心肌細(xì)胞CaMKⅡ活性。結(jié)果:ox-LDL可引起細(xì)胞損傷,具體表現(xiàn)為caspase3/7活性增加,caspase-3的表達(dá)增強(qiáng)以及細(xì)胞活性下降。而APOJ高表達(dá)能夠顯著降低ox-LDL造成的細(xì)胞損傷。ox-LDL能夠增加ROS的產(chǎn)生,具體表現(xiàn)為Nox2/gp91phox和P47表達(dá)升高,而ApoJ可以抑制ox-LDL的表達(dá)。此外,APOJ高表達(dá)能夠降低ox-LDL引起的CaMKⅡ表達(dá),同時(shí)顯著抑制ox-LDL介導(dǎo)的CaMKⅡ活性上調(diào)。CaMKⅡ抑制劑KN93降低ApoJ抗氧化作用,阻礙APOJ對ox-LDL損傷的保護(hù)作用�；钚匝跚宄齽�(ROS)Mn(ⅡI)TBAP也弱化了CaMKⅡ的表達(dá)和ox-LDL誘導(dǎo)的活性增加,并通過減弱ox-LDL誘導(dǎo)的細(xì)胞損傷顯示出與ApoJ類似的結(jié)果。結(jié)論:1.ApoJ可減少ox-LDL誘導(dǎo)的心肌細(xì)胞凋亡,減輕心肌細(xì)胞損傷;2.ox-LDL能利用氧化應(yīng)激,通過ROS/oxCaMKⅡ通路引起細(xì)胞損傷;3.高表達(dá)ApoJ通過抑制ROS/oxCaMKⅡ信號通路降低OX-LDL對心肌細(xì)胞的毒性作用。
[Abstract]:Nowadays, cardiovascular disease has become a major problem restricting people's health. There are many types of cardiovascular diseases, such as ischemic cardiomyopathy, hypertension and atherosclerosis. By investigation, most cardiovascular diseases and as are closely related. People in the industry at home and abroad have never stopped their research on coronary heart disease. According to the latest research results, most unnecessary reactive oxygen speciesros can be stimulated by various factors of coronary heart disease. Reactive oxygen species, in turn, produce pathological processes that induce as. Oxidized low density lipoprotein (LDL) low oxidizednlox-LDLs can not only damage vascular endothelium and induce cytokines, but also induce apoptosis. Apolipoprotein Apolipoprotein-ApoJ (ApoJ) is a new type of multifunctional secretory sugar protein which exists in human tissues and body fluids. It not only participates in the physiological and pathological processes of sperm maturation, tissue remodeling, and complement activation, but also plays an important role in apoptosis and lipid transport. Apolipoprotein J plays a significant role in inhibiting apoptotic cells and thus protects cells from heart damage. But it is worth noting that even though the relationship between apolipoprotein J and ox-LDL plays a decisive role in the growth and death of cardiomyocytes, it has not been paid enough attention to. Aim: to elucidate the effects of ox-LDL on cardiac myocytes and the mechanism of intracellular signal transduction, and to explore whether the high expression of ApoJ can reduce the effect of ox-LDL on neonatal rat ventricular cells in neonatal rats. Methods: adenovirus containing ApoJ gene was used to infect NRVCs to make ApoJ overexpression. CaMK 鈪，

本文編號：1782191