發(fā)布時(shí)間：2018-02-24 19:37

  本文關(guān)鍵詞： 尾加壓素II GPR14 一氧化氮 硫化氫 心梗 心肌細(xì)胞 免疫組化 免疫熒光雙標(biāo) 胱硫醚-γ-裂解酶 蛋白印跡雜交 逆轉(zhuǎn)錄一聚合酶鏈反應(yīng)　出處：《復(fù)旦大學(xué)》2005年博士論文　論文類型：學(xué)位論文

【摘要】：尾加壓素Ⅱ是最早從魚的尾垂體分離出來的一種多肽,最近哺乳動(dòng)物包括人的尾加壓素Ⅱ已經(jīng)被克隆出來。研究表明,尾加壓素Ⅱ在心血管系統(tǒng)有重要作用,它曾被認(rèn)為是最強(qiáng)的縮血管物質(zhì)之一。對于不同種屬的動(dòng)物,靜脈注射尾加壓素Ⅱ產(chǎn)生的心血管反應(yīng)有很大的差異,甚至對于同一種屬的動(dòng)物,報(bào)道也并不一致。尾加壓素Ⅱ能增加人右心房肌的收縮力,但對左心室心肌的直接作用還有待闡明。尾加壓素Ⅱ受體GPR14分布比較廣泛,包括心血管組織,中樞和內(nèi)分泌組織等。人們通過RT-PCR和配體受體結(jié)合實(shí)驗(yàn)證實(shí)GPR14在心血管的表達(dá),但對于GPR14蛋白在心臟的表達(dá)以及它在心臟中分布的具體的細(xì)胞類型還缺乏直接的證據(jù)。另外,尾加壓素Ⅱ可能會(huì)介導(dǎo)心肌細(xì)胞的肥大,在一些慢性缺氧或充血性心衰情況,尾加壓素Ⅱ和其受體出現(xiàn)上調(diào),推測尾加壓素Ⅱ可能參與心血管的重構(gòu)過程。一氧化氮具有舒張血管,保護(hù)心肌的作用。它參與尾加壓素Ⅱ?qū)π难�管作用的調(diào)節(jié)。硫化氫是最近受到關(guān)注的具有生物活性的氣體小分子,能抑制心肌收縮,舒張血管。但它是否參與尾加壓素Ⅱ?qū)π难�管的作�?還需要進(jìn)一步的研究。 我們主要通過免疫組化和免疫熒光雙標(biāo)的方法觀察GPR14在心臟中的表達(dá),結(jié)果表明,GPR14主要表達(dá)于正常大鼠左心室的心肌細(xì)胞上,在心臟的其他部分包括心房,右心室和主動(dòng)脈瓣膜沒有觀察到GPR14蛋白的陽性表達(dá);在心臟中其他的細(xì)胞包括冠脈血管的內(nèi)皮和平滑肌細(xì)胞上未見GPR14的表達(dá)。我們利用Western blot和逆轉(zhuǎn)錄—聚合酶鏈的方法進(jìn)一步證實(shí)GPR14蛋白和基因確實(shí)表達(dá)在左心室。我們又通過免疫組化的方法觀察GPR14蛋白在大鼠心梗后3天和2周梗塞區(qū)邊緣的表達(dá),結(jié)果表明,與假手術(shù)組相比,GPR14在表達(dá)的量和表達(dá)的細(xì)胞類型上,均沒有出現(xiàn)明顯的變化。在房間隔缺損病人的右心房,我們觀察到GPR14受體的表達(dá),主要在靠近心外膜的心肌組織;而在右心耳,GPR14蛋白的表達(dá)與血管分布有關(guān)。 為了從功能學(xué)上進(jìn)一步證實(shí)尾加壓素Ⅱ受體確實(shí)表達(dá)在左心室的心肌上,我們觀察了尾加壓素Ⅱ?qū)ψ笫胰轭^肌的作用,結(jié)果提示,尾加壓素增加左室乳頭肌的收縮強(qiáng)度,但并不改變其他的收縮參數(shù),包括收縮舒張速率,收縮潛伏期,達(dá)到最大收縮張力的時(shí)間以及舒張到一半的時(shí)間。我們還觀察了給麻醉大鼠靜脈注射尾加壓素Ⅱ引起的心血管效應(yīng),主要引起左室收縮壓,股動(dòng)脈收縮壓和舒張壓,心率的下降;心肌收縮力(dp/dt)和心肌舒張能力(-dp/dt)的減弱;左室舒張壓的升高:提示尾加壓素Ⅱ可能對心臟具有抑制作用。我們推測在整體實(shí)驗(yàn)中,尾加壓素Ⅱ可能通過調(diào)節(jié)其他物質(zhì)如一氧化氮和硫化氫等氣體小分子
[Abstract]:Urotensin II is a polypeptide from as early as the tail of a fish pituitary separated, including the tail recently vasopressin II mammals have been cloned. The results showed that urotensin II plays an important role in the cardiovascular system, it was considered to be one of the strongest vasoconstrictor for different species. Animal, intravenous urotensin II the cardiovascular response is very different, even for the same species of animal, reports are not the same. Urotensin II can increase the contractility of right atrium, but the direct effect on left ventricular myocardium still remains to be elucidated. Angiotensin II receptor is widely urotensin the distribution of GPR14, including cardiovascular tissues, central and endocrine tissues. People with GPR14 in cardiovascular experiments confirmed that the expression of RT-PCR through ligand receptor and GPR14 protein in the heart, but for the expression and its distribution in the heart The specific cell types is lack of direct evidence. In addition, urotensin II may be mediated by hypertrophy of cardiomyocytes, in chronic hypoxia or congestive heart failure, urotensin II and its receptor up-regulated, urotensin II that may be involved in cardiovascular remodeling process. Nitric oxide with vasodilator effect, myocardial protection. It is involved in the regulation of urotensin II on cardiovascular effects. Hydrogen sulfide is the recent attention of bioactive molecules, can inhibit myocardial contractility, relaxation of blood vessels. But whether it is involved in the tail function of vasopressin on cardiovascular, further research is needed.
We mainly through immunohistochemistry and double immunofluorescence method to observe expression of GPR14 in the heart, results showed that GPR14 was mainly expressed in the normal left ventricle of rat myocardial cells, in other parts of the heart including atrial, right ventricular and aortic valve was not observed in the positive expression of GPR14 protein in the heart of the other; there was no expression of GPR14 cells including coronary vascular smooth muscle cells and endothelial cells. We use the method of Western blot and reverse transcriptase polymerase chain further confirmed that GPR14 protein and gene expression in the left ventricle. We did observe by immunohistochemistry GPR14 protein in rats after myocardial infarction, 3 days and 2 weeks of infarction zone edge the results show that, compared with the sham operation group, and the expression of GPR14 in the amount of cell types, there was no obvious change. Right atrium in atrial septal defect patients, I The expression of the GPR14 receptor was observed mainly in the cardiac tissue near the epicardium, while in the right auricle, the expression of GPR14 protein was associated with the distribution of blood vessels.
In order to further study on function of urotensin II receptor that has been expressed in the left ventricular myocardium, we observed the effect of urotensin II, on the left ventricular papillary muscle showed that urotensin increased left ventricular papillary muscle contraction strength, but does not change the other parameters including systolic, diastolic and systolic velocity contraction, latency, maximum contraction time and diastolic to half the time. We also observed the anesthetized rat was urotensin II induced cardiovascular effects, mainly caused by left ventricular systolic pressure, systolic arterial pressure and diastolic blood pressure, heart rate decreased; myocardial contractility and diastolic (dp/dt) capacity (-dp/dt) decreased; left ventricular diastolic pressure elevated urotensin II may have inhibitory effects on the heart. We speculate that in vivo, urotensin II may regulate other substances such as nitric oxide Small molecules of gas, such as hydrogen sulfide

【學(xué)位授予單位】：復(fù)旦大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2005
【分類號】：R33

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 李玲,袁文俊,潘秀頡,王偉忠,邱景偉,唐朝樞,唐朝樞;尾加壓素對新生大鼠心肌細(xì)胞一氧化氮合成的影響[J];生理學(xué)報(bào);2002年04期

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本文編號：1531458