大通牦牛心肌組織低氧適應(yīng)發(fā)育學(xué)研究
發(fā)布時(shí)間：2018-12-30 13:49
【摘要】：為探討在高原低氧環(huán)境下不同發(fā)育期大通牦牛心肌組織發(fā)育學(xué)特點(diǎn),本實(shí)驗(yàn)選擇同一海拔梯度不同發(fā)育期(妊娠4月齡、出生1日齡、1月齡、3月齡、6齡月)大通牦牛作為研究對(duì)象,應(yīng)用透射電鏡技術(shù)對(duì)不同發(fā)育期牦牛心肌細(xì)胞線粒體立體形態(tài)學(xué)指標(biāo)進(jìn)行定性和定量分析;運(yùn)用分子生物學(xué)技術(shù)檢測(cè)心肌線粒體標(biāo)志性酶:細(xì)胞色素C氧化酶(Cytochrome C Oxidase,COX)、琥珀酸脫氫酶(Succinatedehydro-genase,SDH)以及低氧誘導(dǎo)因子(Hypoxia-inducible factor,HIF-1α)m RNA進(jìn)行實(shí)時(shí)定量熒光分析;應(yīng)用免疫組織化學(xué)方法對(duì)牦牛心肌組織血管內(nèi)皮因子(vascular endothelial growth factor,VEG)和微血管面密度(Microvessel density,MVD)進(jìn)行檢測(cè)。研究結(jié)果如下:(1)心肌線粒體形態(tài)學(xué)結(jié)果:心肌細(xì)胞內(nèi)線粒體排列整齊均勻,大多呈圓形或卵圓形;外膜、內(nèi)膜完整,膜間隙、嵴和基質(zhì)結(jié)構(gòu)清晰可見,由于分娩及外界環(huán)境的應(yīng)激刺激出生1日齡牦牛心肌細(xì)胞線粒體結(jié)構(gòu)膨大、數(shù)量減少、部分空泡化;心肌線粒體平均截面積、平均體積、面數(shù)密度、體積密度隨年齡的增加呈上升趨勢(shì),差異顯著(P0.05);出生1日齡各項(xiàng)指標(biāo)變化明顯,線粒體平均截面積(AX/μm2)、平均體積(V/μm3)均大于其他年齡段,而面數(shù)密度(Na/μm2)均低于其他年齡組,體積密度(V/μm3)除大于1月齡均小于其他年齡段。(2)心肌組織COX亞基m RNA表達(dá)量結(jié)果:出生1日齡時(shí)牦牛心肌線粒體COX-I、II m RNA表達(dá)量最高;COX-II m RNA表達(dá)量均高于COX-Im RNA。(3)心肌組織SDH亞基m RNA表達(dá)量結(jié)果:心肌組織中SDH 4個(gè)亞基轉(zhuǎn)錄在出生1日齡SDH 4個(gè)亞基表達(dá)量均高于其他年齡段。(4)心肌組織HIF-1αm RNA表達(dá)量結(jié)果:HIF-1αm RNA表達(dá)量隨年齡增長(zhǎng)表達(dá)量穩(wěn)定增加,不同年齡組間差異顯著(P0.05);出生1日齡和6月齡HIF-1αm RNA表達(dá)量顯著高于其他年齡組,差異極顯著(P0.01)。(5)心肌組織VEGF、MVD測(cè)定結(jié)果:不同發(fā)育期大通牦牛心肌中VEGF、MVD均有分布,心肌VEGF、MVD百分比含量隨年齡的增長(zhǎng)而增加,差異顯著(P0.05);出生1日齡VEGF、MVD占單位數(shù)量心肌纖維的比值較其他年齡組含量低。綜上所述,高原低氧環(huán)境下,大通牦牛心肌細(xì)胞線粒體數(shù)目、體積以及心肌組織SDH、COX、HIF-1αm RNA的表達(dá)量、心肌組織VEGF、MVD蛋白百分比含量均隨年齡增長(zhǎng)呈穩(wěn)定上升趨勢(shì)�？赡苡捎诜置浼巴饨绛h(huán)境的應(yīng)激刺激,出生1日齡各項(xiàng)參數(shù)與其他年齡組相比均有明顯變化,線粒體結(jié)構(gòu)受到暫時(shí)性損傷,出生1日齡牦牛在分娩應(yīng)激刺激下,增加心肌組織HIF-1α表達(dá)量和加強(qiáng)調(diào)控線粒體標(biāo)志性酶COX、SDH氧化磷酸化作用使組織細(xì)胞快速?gòu)?fù)氧增加,從而對(duì)心肌組織的供氧供能起到一定的保護(hù)作用。HIF-1α在細(xì)胞核內(nèi)快速積累,進(jìn)而誘導(dǎo)機(jī)體內(nèi)VEGF、MVD因子表達(dá)調(diào)節(jié)氧分壓防止低氧對(duì)細(xì)胞造成損傷。
[Abstract]:In order to study the characteristics of myocardial tissue development in Datong yak at different developmental stages under high altitude hypoxia, we selected the same altitude gradient for different developmental periods (4 months of pregnancy, 1 month of birth, 1 month of birth, 3 months of age). Datong yak was used as the research object. The qualitative and quantitative analysis of mitochondrial stereomorphology of cardiac myocytes of yak at different stages of development was carried out by transmission electron microscopy (TEM). Detection of Myocardial Mitochondrial iconic enzymes (cytochrome C Oxidase (Cytochrome C Oxidase,COX), succinate dehydrogenase (Succinatedehydro-genase,SDH) and hypoxia Induction Factor (Hypoxia-inducible factor,) by Molecular Biological techniques HIF-1 偽) m RNA was used for real-time quantitative fluorescence analysis. Vascular endothelial factor (vascular endothelial growth factor,VEG) and microvessel surface density (Microvessel density,MVD) were measured by immunohistochemical method. The results are as follows: (1) the morphology of myocardial mitochondria: the mitochondria in myocardial cells are arranged neatly and evenly, most of them are round or oval; The outer membrane, intima intact, membrane space, cristae and matrix structure were clearly visible. Due to the stress of labor and external environment, the mitochondrial structure of 1 day old yak cardiomyocytes was enlarged, the number of mitochondria was reduced, and part of vacuolation was observed. The mean sectional area, mean volume, face number density and volume density of myocardial mitochondria increased with age (P0.05). At 1 day of birth, the changes of the indexes were obvious. The average mitochondrial area (AX/ 渭 m 2) and mean volume (V / 渭 m 3) were higher than those of other age groups, but the Na/ 渭 m 2 was lower than that of other age groups. The volume density (V / 渭 m 3) was lower than that of other age groups except over 1 month. (2) the expression of COX subunit m RNA in myocardium was the highest in yak myocardial mitochondria at 1 day of life. The expression of COX-II m RNA was higher than that of COX-Im RNA. (3) the expression of SDH subunit m RNA in myocardial tissue was higher than that of other age groups. 4) the expression of HIF-1 偽 m RNA in myocardium: the expression of HIF-1 偽 m RNA increased steadily with age. There were significant differences among different age groups (P0.05). The expression of HIF-1 偽 m RNA at 1 day and 6 months of birth was significantly higher than that of other age groups, and the difference was very significant (P0.01). (5). The results of VEGF,MVD analysis showed that there was a distribution of VEGF,MVD in myocardium of Datong yak at different developmental stages, and the expression of VEGF,MVD in myocardium of Datong yak was significantly higher than that of other age groups. The percentage of MVD increased with age (P0.05). The ratio of VEGF,MVD per unit number of myocardial fibers at 1 day old was lower than that in other age groups. In conclusion, the number and volume of mitochondria, the expression of SDH,COX,HIF-1 偽 m RNA and the percentage of VEGF,MVD protein in myocardium of Datong yak increased steadily with age under high altitude hypoxia. It is possible that the parameters of 1 day old of birth are obviously changed compared with other age groups due to the stress stimulation of labor and external environment. The mitochondrial structure is temporarily damaged, and the 1 day old yak is stimulated by labor stress. Increasing the expression of HIF-1 偽 in myocardium and regulating the oxidative phosphorylation of mitochondrial iconic enzyme COX,SDH increased the rapid reoxygenation of tissue cells. HIF-1 偽 accumulates rapidly in the nucleus, and then induces the expression of VEGF,MVD factor to regulate the partial pressure of oxygen to prevent the damage to cells caused by hypoxia.
【學(xué)位授予單位】：青海大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：S823.85
本文編號(hào)：2395663
[Abstract]:In order to study the characteristics of myocardial tissue development in Datong yak at different developmental stages under high altitude hypoxia, we selected the same altitude gradient for different developmental periods (4 months of pregnancy, 1 month of birth, 1 month of birth, 3 months of age). Datong yak was used as the research object. The qualitative and quantitative analysis of mitochondrial stereomorphology of cardiac myocytes of yak at different stages of development was carried out by transmission electron microscopy (TEM). Detection of Myocardial Mitochondrial iconic enzymes (cytochrome C Oxidase (Cytochrome C Oxidase,COX), succinate dehydrogenase (Succinatedehydro-genase,SDH) and hypoxia Induction Factor (Hypoxia-inducible factor,) by Molecular Biological techniques HIF-1 偽) m RNA was used for real-time quantitative fluorescence analysis. Vascular endothelial factor (vascular endothelial growth factor,VEG) and microvessel surface density (Microvessel density,MVD) were measured by immunohistochemical method. The results are as follows: (1) the morphology of myocardial mitochondria: the mitochondria in myocardial cells are arranged neatly and evenly, most of them are round or oval; The outer membrane, intima intact, membrane space, cristae and matrix structure were clearly visible. Due to the stress of labor and external environment, the mitochondrial structure of 1 day old yak cardiomyocytes was enlarged, the number of mitochondria was reduced, and part of vacuolation was observed. The mean sectional area, mean volume, face number density and volume density of myocardial mitochondria increased with age (P0.05). At 1 day of birth, the changes of the indexes were obvious. The average mitochondrial area (AX/ 渭 m 2) and mean volume (V / 渭 m 3) were higher than those of other age groups, but the Na/ 渭 m 2 was lower than that of other age groups. The volume density (V / 渭 m 3) was lower than that of other age groups except over 1 month. (2) the expression of COX subunit m RNA in myocardium was the highest in yak myocardial mitochondria at 1 day of life. The expression of COX-II m RNA was higher than that of COX-Im RNA. (3) the expression of SDH subunit m RNA in myocardial tissue was higher than that of other age groups. 4) the expression of HIF-1 偽 m RNA in myocardium: the expression of HIF-1 偽 m RNA increased steadily with age. There were significant differences among different age groups (P0.05). The expression of HIF-1 偽 m RNA at 1 day and 6 months of birth was significantly higher than that of other age groups, and the difference was very significant (P0.01). (5). The results of VEGF,MVD analysis showed that there was a distribution of VEGF,MVD in myocardium of Datong yak at different developmental stages, and the expression of VEGF,MVD in myocardium of Datong yak was significantly higher than that of other age groups. The percentage of MVD increased with age (P0.05). The ratio of VEGF,MVD per unit number of myocardial fibers at 1 day old was lower than that in other age groups. In conclusion, the number and volume of mitochondria, the expression of SDH,COX,HIF-1 偽 m RNA and the percentage of VEGF,MVD protein in myocardium of Datong yak increased steadily with age under high altitude hypoxia. It is possible that the parameters of 1 day old of birth are obviously changed compared with other age groups due to the stress stimulation of labor and external environment. The mitochondrial structure is temporarily damaged, and the 1 day old yak is stimulated by labor stress. Increasing the expression of HIF-1 偽 in myocardium and regulating the oxidative phosphorylation of mitochondrial iconic enzyme COX,SDH increased the rapid reoxygenation of tissue cells. HIF-1 偽 accumulates rapidly in the nucleus, and then induces the expression of VEGF,MVD factor to regulate the partial pressure of oxygen to prevent the damage to cells caused by hypoxia.
【學(xué)位授予單位】：青海大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：S823.85
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