【摘要】：缺血預(yù)處理(ischemic preconditioning, IPC)對(duì)缺血/再灌注心肌的保護(hù)作用已在多種屬動(dòng)物在體、離體心臟和心肌細(xì)胞上得到證實(shí),該現(xiàn)象可以被缺氧預(yù)處理(hypoxic preconditioning, HPC)模擬。預(yù)處理的保護(hù)作用具有時(shí)相性,包括預(yù)處理后即刻出現(xiàn)并持續(xù)1-3小時(shí)的早期保護(hù)和12-24小時(shí)再度出現(xiàn)并持續(xù)24-72小時(shí)的延遲保護(hù),其中延遲保護(hù)作用具有更重要的臨床意義。從預(yù)處理心肌保護(hù)機(jī)制的研究入手,探討缺血性心臟病防治的新策略,受到基礎(chǔ)和臨床醫(yī)學(xué)界的高度重視。 現(xiàn)有研究結(jié)果表明,預(yù)處理的延遲保護(hù)機(jī)理涉及腺苷、內(nèi)皮源性舒張因子(EDRF)、一氧化氮(NO)等內(nèi)源性保護(hù)介質(zhì)釋放,蛋白激酶C(protein kinase C, PKC)和絲裂素活化蛋白激酶家系(mitogen-activated protein kinases, MAPKs)等細(xì)胞信號(hào)轉(zhuǎn)導(dǎo)途徑激活,以及內(nèi)源性保護(hù)蛋白轉(zhuǎn)錄、合成、翻譯后修飾改變等多種因素,其中內(nèi)源性保護(hù)蛋白合成上調(diào)是延遲保護(hù)的關(guān)鍵環(huán)節(jié),但目前預(yù)處理延遲保護(hù)過程中心肌蛋白質(zhì)組學(xué)變化及其細(xì)胞信號(hào)轉(zhuǎn)導(dǎo)機(jī)制尚未完全闡明。 本工作證實(shí)HPC對(duì)乳鼠心肌細(xì)胞缺氧/復(fù)氧(hypoxia/reoxygenation,H/R)損傷具有延遲保護(hù)作用的基礎(chǔ)上,進(jìn)一步研究HPC延遲心肌保護(hù)的細(xì)胞信號(hào)轉(zhuǎn)導(dǎo)機(jī)制及其誘導(dǎo)的心肌細(xì)胞蛋白質(zhì)組學(xué)變化。具體方法如下: 1、在乳鼠心肌細(xì)胞H/R模型上,采用臺(tái)盼藍(lán)排斥實(shí)驗(yàn)檢測(cè)心肌細(xì)胞存活率,以TUNEL法、DNA特異性熒光素染料Hoechst33258染色等方法測(cè)定心肌細(xì)胞凋亡率,證實(shí)缺氧預(yù)處理明顯減輕24小時(shí)后H/R誘導(dǎo)的心肌細(xì)胞損傷。 2、Western bolt檢測(cè)證實(shí),HPC后24h心肌細(xì)胞內(nèi)PKC亞型nPKCε和MAPKs家系成員細(xì)胞外信號(hào)調(diào)節(jié)激酶(extracellular signal-regulated
[Abstract]:The protective effect of ischemic preconditioning (ischemic preconditioning, IPC) on ischemic / reperfusion myocardium has been confirmed in many species of animals in vivo. This phenomenon can be simulated by hypoxic preconditioning (hypoxic preconditioning, HPC) in isolated hearts and cardiomyocytes. The protective effects of preconditioning are temporal, including the early protection which appears immediately after pretreatment and lasts for 1-3 hours, and the delayed protection which reappears at 12-24 hours and lasts for 24-72 hours, among which the delayed protection has more important clinical significance. Based on the study of myocardial protection mechanism of preconditioning, a new strategy for prevention and treatment of ischemic heart disease has been paid great attention to in the field of basic and clinical medicine. The results show that the delayed protection mechanism of pretreatment involves the release of endogenous protective mediums, such as adenosine, endothelium-derived diastolic factor (EDRF), nitric oxide (NO), and so on. Protein kinase C (protein kinase C, PKC) and mitogen-activated protein kinase family (mitogen-activated protein kinases, MAPKs) and other cell signal transduction pathway activation, as well as endogenous protection protein transcription, synthesis, post-translational modification and other factors, The up-regulation of endogenous protection protein synthesis is the key link of delayed protection, but the proteomics changes and the mechanism of cell signal transduction in the process of pretreatment delayed protection have not been fully elucidated. Based on the conclusion that HPC has delayed protection against hypoxia / reoxygenation (H / R) injury in neonatal rat cardiomyocytes, the mechanism of cellular signal transduction and the proteomic changes of cardiomyocytes induced by HPC delayed myocardial protection were further studied. The specific methods are as follows: 1. The survival rate of neonatal rat cardiomyocytes was determined by trypan blue rejection test on the H / R model. The apoptotic rate of cardiomyocytes was determined by TUNEL staining and Hoechst33258 staining with DNA-specific fluorescein dye. The results showed that hypoxia preconditioning significantly alleviated the cardiomyocyte injury induced by H / R after 24 hours. 2Western bolt analysis confirmed the formation of nPKC 蔚 and MAPKs subtypes of PKC subtype in cardiomyocytes 24 hours after HPC. Extracellular signal regulated kinase (extracellular signal-regulated)
【學(xué)位授予單位】：中國(guó)人民解放軍軍醫(yī)進(jìn)修學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2005
【分類號(hào)】：R363

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