【摘要】：第1章引言急性心肌梗塞在心血管疾病中發(fā)病率和死亡率極高,缺血/再灌注損傷仍是急性心肌梗塞臨床治療中面臨的難題。如何減輕缺血/再灌注損傷是心肌損傷與保護(hù)的研究熱點(diǎn)。研究心肌損傷發(fā)生機(jī)制,將有助于我們?yōu)榕R床防治心肌缺血損傷提供理論和實(shí)踐依據(jù)。因此,我們基于lncRNA-miRNA-Notch1調(diào)控假說(shuō),建立心肌細(xì)胞缺氧/復(fù)氧損傷及自噬保護(hù)模型,通過(guò)高通量測(cè)序技術(shù)及生物信息學(xué)比較分析,篩選出在自噬保護(hù)中具有調(diào)控Notch1蛋白作用的miRNA,以及具有調(diào)控該miRNA的lncRNA,為研究Notch1信號(hào)通路在自噬性心肌保護(hù)的分子機(jī)制提供科學(xué)依據(jù)。第2章Notch1信號(hào)通路增強(qiáng)自噬對(duì)抗心肌缺氧復(fù)氧損傷的保護(hù)作用目的:篩選有效調(diào)控自噬的3-MA及RAPA試劑濃度,以及探討Notch1信號(hào)通路在自噬對(duì)抗心肌缺氧復(fù)氧損傷中的作用。方法:建立心肌細(xì)胞損傷模型,給予3-MA、RAPA干預(yù),檢測(cè)細(xì)胞活力,篩選3-MA及RAPA有效試劑濃度;利用Ad-N1ICD及Ad-N1ICD-shRNA干預(yù)Notch1信號(hào)通路,檢測(cè)H/R損傷中心肌細(xì)胞活力情況。結(jié)果:本研究中3-MA和RAPA最佳試劑濃度分別為1μM和50nM;Notch1過(guò)表達(dá)可提高H/R的心肌細(xì)胞活力(p0.01),干擾Notch1可明顯降低H/R心肌細(xì)胞活力(p0.05);3-MA加入Notch1過(guò)表達(dá)組中,H/R的心肌細(xì)胞活力再次降低(p0.01),RAPA加入Notch1干擾組中,H/R的心肌細(xì)胞活力恢復(fù)(p0.01)。結(jié)論:Notch1信號(hào)通路可增強(qiáng)自噬對(duì)抗心肌缺氧復(fù)氧損傷發(fā)揮保護(hù)作用。第3章高通量測(cè)序及生物信息學(xué)篩選自噬性心肌保護(hù)中Notch1信號(hào)通路相關(guān)miRNA及l(fā)ncRNA目的:借助高通量測(cè)序及生物信息學(xué)比較分析篩選在自噬性心肌保護(hù)中具有調(diào)控Notch1信號(hào)通路的關(guān)鍵miRNA及l(fā)ncRNA。方法:提取心肌缺氧/復(fù)氧損傷中的總RNA,借助高通量測(cè)序及生物信息學(xué)分析系統(tǒng),篩選具有差異表達(dá)的miRNA及l(fā)ncRNA。結(jié)果:篩選出了可能調(diào)控Notch1基因表達(dá)的miR-702-5p、miR-344g、miR-323-5p、miR-6334、miR-5132-5p、miR-15b-5p、miR-3562;生物信息學(xué)分析發(fā)現(xiàn)lncRNA-4321可與miR-702-5p直接結(jié)合并影響其活性;在RAPA和3-MA處理心肌細(xì)胞中,Realtime-PCR檢測(cè)顯示lncRNA-4321表達(dá)量與心肌自噬成正比,miR-702-5p表達(dá)量與心肌自噬成反比。結(jié)論:lncRNA-4321/miR-702-5p/Notch1可能在自噬性心肌保護(hù)中發(fā)揮重要作用。
[Abstract]:Chapter 1 introduces that the incidence and mortality of acute myocardial infarction in cardiovascular disease is very high. Ischemia / reperfusion injury is still a difficult problem in the clinical treatment of acute myocardial infarction. How to reduce ischemia / reperfusion injury is the focus of myocardial injury and protection. Studying the mechanism of myocardial injury will help us to provide theoretical and practical basis for clinical prevention and treatment of myocardial ischemia injury. Therefore, based on the lncRNA-miRNA-Notch1 regulation hypothesis, we established a cardiomyocyte model of hypoxia / reoxygenation injury and autophagy protection, and analyzed it by high-throughput sequencing and bioinformatics. The screening of miRNA, which has the function of regulating Notch1 protein in autophagy protection and lncRNA, which has the function of regulating miRNA provide scientific basis for studying the molecular mechanism of Notch1 signaling pathway in autophagy myocardial protection. Chapter 2 Notch1 signaling pathway enhances the protective effect of autophagy on myocardial hypoxic-reoxygenation injury objective: to screen the concentration of 3-MA and RAPA reagents that can effectively regulate autophagy and to explore the role of Notch1 signaling pathway in anti-hypoxia and reoxygenation injury of myocardium. Methods: myocardial injury model was established, 3-MARAPA was given to detect cell viability, 3-MA and RAPA effective reagent concentrations were screened, and Notch1 signaling pathway was interfered by Ad-N1ICD and Ad-N1ICD-shRNA to detect myocardial cell viability in H / R injury. Results: the best concentration of 3-MA and RAPA in this study was 1 渭 M and 50nM respectively, which increased the viability of H / R cardiomyocytes (p0.01). Interfering with Notch1 could significantly reduce the activity of H / R cardiomyocytes (p0.05) 3-MA added to Notch1 overexpression group. The secondary decrease (p0.01) and the activity recovery of H / R in the Notch1 interference group (p0.01). Conclusion the fraction Notch1 signaling pathway can enhance the protective effect of autophagy on myocardial hypoxia and reoxygenation injury. Chapter 3 High-throughput sequencing and Bioinformatics screening Notch1 signaling Path-Related miRNA and lncRNA in autophagy Myocardial Protection objective: to regulate Notch1 in autophagic myocardial protection by high-throughput sequencing and bioinformatics comparative analysis Key miRNA and lncRNA. of signal Pathway Methods: total RNA, from myocardial hypoxia / reoxygenation injury was extracted by high throughput sequencing and bioinformatics analysis system to screen miRNA and lncRNA. with differential expression. Results: the bioinformatics analysis of miR-702-5p,miR-344g,miR-323-5p,miR-6334,miR-5132-5p,miR-15b-5p,miR-3562;, which could regulate the expression of Notch1 gene, found that lncRNA-4321 could directly bind to miR-702-5p and affect its activity. In RAPA and 3-MA treated cardiomyocytes, the expression of lncRNA-4321 was directly proportional to myocardial autophagy and the expression of miR-702-5p was inversely proportional to myocardial autophagy. Conclusion: 1: lncRNA-4321% miR-702-5p / Notch1 may play an important role in the protection of autophagic myocardium.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R54

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 張麗;夏德雨;李國(guó)輝;蔣姍姍;谷芳娜;梁英民;;Notch信號(hào)在急性早幼粒細(xì)胞白血病患者骨髓中的表達(dá)[J];第四軍醫(yī)大學(xué)學(xué)報(bào);2009年24期

2 姜萌;何奔;;Notch信號(hào)對(duì)血管新生調(diào)控作用的研究進(jìn)展[J];上海交通大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2010年05期

3 ;Effect of Trastuzumab on Notch-1 Signaling Pathway in Breast Cancer SK-BR3 Cells[J];Chinese Journal of Cancer Research;2012年03期

4 魯茁壯,王立生,吳祖澤;Notch信號(hào)通路研究進(jìn)展[J];生理科學(xué)進(jìn)展;2004年02期

5 張謙慎;Notch信號(hào)與肺發(fā)育[J];中國(guó)當(dāng)代兒科雜志;2004年02期

6 孫力,詹啟敏,章文華;Notch信號(hào)通路與腫瘤[J];國(guó)外醫(yī)學(xué)(腫瘤學(xué)分冊(cè));2004年09期

7 張謙慎,常立文,劉漢楚,容志惠,祝華平,陳紅兵,李文斌;Notch信號(hào)與大鼠肺發(fā)育的關(guān)系研究[J];華中科技大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2004年03期

8 張謙慎,常立文,劉漢楚,容志惠,陳紅兵;TEMPORAL EXPRESSION OF NOTCH RECEPTORS DURING LUNG DEVELOPMENT IN RAT[J];Journal of Shanghai Second Medical University;2005年02期

9 吳穎亞;王季石;;Notch及其配體在血液系統(tǒng)中的作用[J];國(guó)外醫(yī)學(xué).輸血及血液學(xué)分冊(cè);2005年05期

10 張謙慎,常立文,劉漢楚,容志惠,陳紅兵;Relationship between Notch Receptors and Hyperoxia-induced Lung Injury in Newborn Rats[J];華中科技大學(xué)學(xué)報(bào)(醫(yī)學(xué)英德文版);2005年02期

相關(guān)會(huì)議論文 前10條

1 張麗;梁英民;夏德雨;李國(guó)輝;蔣姍姍;谷芳娜;;Notch信號(hào)在急性早幼粒細(xì)胞白血病患者骨髓中的表達(dá)研究[A];第12屆全國(guó)實(shí)驗(yàn)血液學(xué)會(huì)議論文摘要[C];2009年

2 ;Expression profile of Notch-related genes in multi-drug resistant K562/A02 cells compared with parental K562 cells[A];第12屆全國(guó)實(shí)驗(yàn)血液學(xué)會(huì)議論文摘要[C];2009年

3 ;Notch signaling in lymphopoiesis[A];第10屆全國(guó)實(shí)驗(yàn)血液學(xué)會(huì)議論文摘要匯編[C];2005年

4 韓驊;;Notch信號(hào)途徑的功能和作用機(jī)理的研究[A];全面建設(shè)小康社會(huì)：中國(guó)科技工作者的歷史責(zé)任——中國(guó)科協(xié)2003年學(xué)術(shù)年會(huì)論文集（下）[C];2003年

5 趙昱;侯麗宏;馬磊;朱正華;朱蕭玲;王強(qiáng);呂巖;陳紹洋;;Electroacupuncture pretreatment induces the tolerance against focal cerebral ischemia through activation of Notch pathway[A];中華醫(yī)學(xué)會(huì)第五次全國(guó)重癥醫(yī)學(xué)大會(huì)論文匯編[C];2011年

6 黃佳圓;王銳;陳龍邦;;Expression of Notch-1 and Its Clinical Significance in Different Histological Subtypes of Human Lung Adenocarcinoma[A];2013華東胸部腫瘤論壇暨第六屆浙江省胸部腫瘤論壇論文集[C];2013年

7 ;Aberrant expression profile of Notch signaling pathway involved in immune thrombocytopenic purpura patients[A];第12屆全國(guó)實(shí)驗(yàn)血液學(xué)會(huì)議論文摘要[C];2009年

8 ;Notch induced protein degradation in lymphoid development[A];第六屆全國(guó)免疫學(xué)學(xué)術(shù)大會(huì)論文集[C];2008年

9 ;The functions of ptrl in Notch and Integrin pathways[A];2012全國(guó)發(fā)育生物學(xué)大會(huì)摘要集[C];2012年

10 Yaochun Wang;Guorui Dou;Lin Wang;Hua Han;;Notch signaling:licenses and limits cellular responses to extrinsic stimulations?[A];中國(guó)生物化學(xué)與分子生物學(xué)會(huì)第十屆會(huì)員代表大會(huì)暨全國(guó)學(xué)術(shù)會(huì)議摘要集[C];2010年

相關(guān)重要報(bào)紙文章 前1條

1 ;為什么心在左肝在右？[N];新華每日電訊;2004年

相關(guān)博士學(xué)位論文 前10條

1 王凱;Notch信號(hào)在小鼠創(chuàng)傷性腦損傷后神經(jīng)元死亡中的作用及機(jī)制研究[D];第四軍醫(yī)大學(xué);2015年

2 陳娟娟;Notch重組配體D1R在促進(jìn)造血重建中的作用與機(jī)理[D];第四軍醫(yī)大學(xué);2015年

3 梁燕;Notch3對(duì)低氧誘導(dǎo)的人肺動(dòng)脈平滑肌細(xì)胞增殖的作用機(jī)制探討[D];首都醫(yī)科大學(xué);2014年

4 孫建華;肺癌患者在乏氧條件下VEGF和Dll4/Notch通路分子的異常表達(dá)和相互關(guān)系[D];山東大學(xué);2015年

5 丁君;缺氧誘導(dǎo)因子1-α和Notch1的關(guān)系及其在阿爾茨海默病中的研究[D];華中科技大學(xué);2015年

6 付偉;Notch信號(hào)在慢性粒細(xì)胞白血病細(xì)胞增殖和伊馬替尼耐藥的調(diào)控作用研究[D];第四軍醫(yī)大學(xué);2014年

7 柏振江;Notch配體DLL4在手足口病患兒中表達(dá)、臨床意義和功能分析[D];蘇州大學(xué);2016年

8 田z，

本文編號(hào)：2214067