發(fā)布時間：2018-05-14 21:17

  本文選題：牦牛 + 心臟��； 參考：《甘肅農(nóng)業(yè)大學(xué)》2016年博士論文

【摘要】：牦牛是一種長期生活在海拔3000m以上高寒低氧地區(qū)的特殊物種,具有“高原之舟”的美稱。在長期的生活狀態(tài)下,牦牛在其生理、生化和基因方面都取得了穩(wěn)定的遺傳特性。已有研究證明,動物生活在低氧環(huán)境中容易引發(fā)高山病,最明顯的是肺動脈高壓和右心室肥厚,其中以�？苿游镒顬槊黠@。目前,課題組已經(jīng)對牦牛肺臟血管進(jìn)行了大量研究,證明確實有自己的特點。目前牦牛心臟血管方面的研究主要集中在毛細(xì)血管床,牦牛心臟動脈和靜脈及冠狀動脈的解剖結(jié)構(gòu)的研究方面。雖對冠狀動脈進(jìn)行了大量的研究,但年齡段劃分不夠細(xì)致,研究只停留在解剖結(jié)構(gòu)的研究,也未見相關(guān)因子在血管和心肌組織上表達(dá)情況的報道。因此,本研究選取32粒牦牛心臟(1日齡、6月齡、1歲、2歲和5歲牦牛)作為研究對象,應(yīng)用免疫組織化學(xué)技術(shù)、免疫電鏡技術(shù)和Elisa檢測技術(shù)對牦牛心肌組織結(jié)構(gòu)和HIF-1a、VEGF、HSP27和HSP70蛋白在牦牛冠狀動脈上的表達(dá)進(jìn)行了定位和定量的研究,以期為心臟對低氧環(huán)境的適應(yīng)機(jī)理提供理論依據(jù),為高原醫(yī)學(xué)的發(fā)展提供基礎(chǔ)數(shù)據(jù)。結(jié)果顯示:1.在牦牛不同的年齡階段,心肌細(xì)胞超微結(jié)構(gòu)呈現(xiàn)了規(guī)律性的變化,主要體現(xiàn)在線粒體、細(xì)胞膜和膠原纖維等方面。2.6月齡牦牛心臟內(nèi)膠原纖維直徑最大。隨著年齡的增加,牦牛心臟內(nèi)Ⅰ型膠原纖維數(shù)量逐漸增多。5歲時,牦牛心臟內(nèi)以Ⅰ型膠原纖維占優(yōu)勢。3.在不同的年齡階段,線粒體形態(tài)發(fā)生變化。1日齡牦牛心臟中線粒體聚集在核的周圍,形態(tài)較圓。從5月齡至2歲,線粒體形態(tài)呈橢圓形,分散在肌原纖維之間,線粒體的嵴排列整齊。在5歲牦牛中心臟中出現(xiàn)矩形線粒體,嵴變得不規(guī)則。線粒體密度和活性的變化:線粒體復(fù)合物酶ⅠⅡ和Ⅴ的活性隨年齡增大,而復(fù)合物酶Ⅲ和Ⅳ的活性從1日齡增大至1歲,之后逐漸下降。4.HIF-1a主要在冠脈的內(nèi)皮細(xì)胞和心肌細(xì)胞有表達(dá),在冠脈平滑肌細(xì)胞中未見表達(dá)。VEGF在內(nèi)皮細(xì)胞和冠脈的平滑肌細(xì)胞中有表達(dá),在心肌細(xì)胞中未見表達(dá)�？瞻讓φ罩形匆娺@兩種蛋白表達(dá)。免疫電鏡結(jié)果顯示:HIF-1a蛋白在內(nèi)皮細(xì)胞的胞核(弱信號)和胞質(zhì)(強(qiáng)信號)表達(dá),在心肌細(xì)胞胞質(zhì)中有強(qiáng)信號表達(dá),在平滑肌細(xì)胞上未見表達(dá)。VEGF蛋白在冠狀動脈內(nèi)皮細(xì)胞胞質(zhì)(強(qiáng)信號)和胞核(弱信號)表達(dá),在冠脈平滑肌細(xì)胞的胞核及核周表達(dá)。5.HIF-1a的分泌量在心肌組織和冠脈中,從1日齡開始一直增加至2歲,之后又下降。在1日齡和2歲之間的分泌量差異顯著。然而,VEGF蛋白在心肌和冠脈中的分泌量隨年齡增大持續(xù)增多。HIF-1a和VEGF的分泌量均左心室高于右心室,前降支高于左旋支。6.HSP27和HSP70蛋白在冠狀動脈血管內(nèi)皮細(xì)胞、心肌細(xì)胞的胞質(zhì)中表達(dá)較強(qiáng)。HSP70蛋白還在中膜的平滑肌細(xì)胞核中強(qiáng)表達(dá)。HSP27基因在各個年齡段均左心室的表達(dá)量最高,其次是右心室、左心房和右心房。HSP70基因從6月齡開始左心室表達(dá)量較高。HSP27和HSP70基因的表達(dá)量均是心室高于心房。
[Abstract]:Yak is a special species living in alpine and hypoxic areas above 3000m altitude for a long time. It has the beauty of the "boat of Plateau". In the long life, yaks have obtained stable genetic characteristics in its physiological, biochemical and genetic aspects. At present, the research group has done a lot of research on the lung blood vessels of yak, which proved to have its own characteristics. At present, the research on the heart vessels of yaks is mainly focused on the capillary bed, the heart artery of yak, and the anatomical structure of the vein and coronary artery. Although a large number of studies have been carried out on the coronary artery, the age division is not meticulous, the study only stays in the study of anatomical structure, and there is no related factors in the expression of blood vessels and myocardium. Therefore, this study selected 32 yak hearts (1 days of age, 6 month old, 1 years, 2 and 5 Yyears) as the research object, The structure of yak myocardial tissue and the expression of HIF-1a, VEGF, HSP27 and HSP70 protein on the coronary artery of yak were studied by immuno histochemistry, immuno electron microscopy and Elisa detection, in order to provide a theoretical basis for the adaptation mechanism of the heart to hypoxia environment and provide the basis for the development of plateau medicine. The results showed that: 1. the ultrastructure of cardiac myocytes in the different age of yak showed a regular change, mainly in the mitochondria, cell membrane and collagen fiber, the diameter of collagen fiber in the heart of yak was the largest in.2.6 month old yak. With the increase of age, the number of type I collagen fibers in the yak heart gradually increased.5 years old, yak In the heart of the cattle, the type I collagen fiber occupies the dominant.3. in the different age stages. The mitochondrial morphogenesis changes in the heart of.1 yak. The mitochondria are round and round in the heart of the yak heart. From 5 month old to 2 years old, the mitochondria are elliptical in shape, scattered in the myofibrils, the ridge of the grain body is arranged neatly. It appears in the center of the yak at the age of 5. The mitochondrial density and activity change: the activity of mitochondrial complex enzyme I II and V increases with age, and the activity of complex enzyme III and IV increases from 1 days to 1 years old, and then.4.HIF-1a decreases mainly in the endothelial cells and cardiac myocytes in the coronary artery and is not found in the coronary smooth muscle cells. The expression of.VEGF was expressed in the smooth muscle cells of endothelial cells and coronary arteries, and no expression was found in the cardiomyocytes. No expression of these two proteins was found in the blank control. The results of immuno electron microscopy showed that the expression of HIF-1a protein in the nucleus (weak signal) and cytoplasm (strong signal) of the endothelial cells was strongly expressed in the cytoplasm of the cardiac myocytes, and the smooth muscle was fine in the smooth muscle cells. The expression of.VEGF protein in the cytoplasm (strong signal) and nucleus (weak signal) of the coronary artery endothelial cells was not expressed on the cell. The secretion of.5.HIF-1a expressed in the nucleus and peri of the coronary artery smooth muscle cells in the myocardial tissue and coronary artery, from 1 days of age to 2 years, and then decreased. The difference of secretion between 1 days and 2 years old was significant. However, the secretion of VEGF protein in the myocardium and coronary artery increased with age and the secretion of.HIF-1a and VEGF was higher than that of the right ventricle. The anterior descending branch was higher than the left branch.6.HSP27 and the HSP70 protein in the coronary vascular endothelial cells. The strong.HSP70 protein expressed in the cytoplasm of the cardiac myocytes was strong in the nucleus of the smooth muscle of the middle membrane. The expression of.HSP27 gene in the left ventricle was the highest at all ages, followed by the right ventricle, the left atrium and the right atrium, the.HSP70 gene was higher in the left ventricle from 6 month old and the expression of the HSP70 gene in the left ventricle was higher than that of the atrium.

【學(xué)位授予單位】：甘肅農(nóng)業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：S823.85

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