奶牛脂肪肝綜合診斷指標(biāo)的篩選及氧化應(yīng)激對犢牛肝臟線粒體OCT活性的影響
發(fā)布時(shí)間：2018-07-17 20:38
【摘要】：奶牛脂肪肝是營養(yǎng)代謝性疾病之一,是由于奶牛肝脂代謝發(fā)生障礙,導(dǎo)致過量的脂類以甘油三脂(TG)的形式在肝臟蓄積,同時(shí)伴有嚴(yán)重的肝臟功能紊亂。奶牛脂肪肝發(fā)病率高達(dá)50%以上,患牛表現(xiàn)出明顯的產(chǎn)奶量下降以及繁殖力和免疫力的降低,嚴(yán)重阻礙奶牛業(yè)的發(fā)展。因此,及時(shí)對奶牛脂肪肝進(jìn)行確診顯得尤為重要。目前,確診脂肪肝只能依據(jù)肝組織活檢,但因該方法對操作者技能要求較高,影響受檢奶牛的生產(chǎn)性能,使得在生產(chǎn)實(shí)踐中的可操作性和可行性差。為了方便、準(zhǔn)確的診斷奶牛脂肪肝,本研究探討奶牛脂肪肝的臨床綜合診斷手段以及發(fā)現(xiàn)可能的診斷指標(biāo)。另有報(bào)道,高脂會(huì)引發(fā)奶牛肝臟線粒體氧化應(yīng)激,而肝臟線粒體特異性酶鳥氨酸氨基甲酰轉(zhuǎn)移酶(OCT)在脂肪肝患牛血清中升高,但線粒體氧化應(yīng)激與線粒體OCT活性變化的關(guān)系尚不清楚。本研究以體外分離的犢牛肝臟線粒體氧化損傷為模型,探討線粒體氧化損傷對OCT活性的影響。結(jié)果如下:1.采用主成分分析和聚類分析兩種數(shù)學(xué)統(tǒng)計(jì)分析方法,結(jié)合臨床生產(chǎn)實(shí)際,對112頭奶牛血漿中與肝臟疾病相關(guān)的生化指標(biāo)天門冬氨酸氨基轉(zhuǎn)移酶(AST)、總膽紅素(TBIL)、TG、鈣(Ca)、鉀(K+)、非酯化脂肪酸(NEFA)、OCT和脂多糖(LPS)進(jìn)行分析,并以超聲波檢查和體況評分作為輔助手段,共同研究奶牛脂肪肝可能的臨床綜合診斷指標(biāo)。結(jié)果顯示:112頭受檢奶�？煞譃槿齻�(gè)組,疑似脂肪肝組奶牛13頭,其中產(chǎn)后奶牛占84%;健康組奶牛7頭;潛在發(fā)病組奶牛92頭。圍產(chǎn)期奶牛中疑似脂肪肝奶牛發(fā)病率占圍產(chǎn)期奶牛的44%。血漿NEFA、OCT、TBIL、K+、AST以及LPS水平升高,TG和Ca水平降低。疑似脂肪肝組B超檢查有10頭奶牛其肝臟可見液性暗區(qū)和疑似鈣化灶的亮點(diǎn),疑似脂肪肝。疑似脂肪肝組奶牛的體況評分均值為4.25,分值偏高。2.利用Fenton反應(yīng)的Fe2+/H2O2體系,體外構(gòu)建犢牛肝臟線粒體氧化損傷模型,檢測線粒體內(nèi)OCT的活性,研究犢牛肝臟線粒體氧化損傷對OCT活性的影響。結(jié)果顯示:隨著H2O2濃度的增加,線粒體丙二醛(MDA)含量以及線粒體腫脹程度增加,線粒體內(nèi)琥珀酸脫氫酶(SDH)和OCT活性降低,且在一定范圍內(nèi)呈濃度依賴性關(guān)系。自由基清除劑甘露醇可以顯著抑制Fenton反應(yīng)條件下誘導(dǎo)的線粒體MDA含量和線粒體腫脹程度的增加以及線粒體內(nèi)SDH活性的降低。在Fenton反應(yīng)條件下,體外成功構(gòu)建犢牛肝臟線粒體氧化損傷模型,在高濃度的Fe2+/H2O2反應(yīng)體系中,線粒體內(nèi)OCT活性呈極顯著降低。從以上結(jié)果可得出以下結(jié)論:1.血漿生化指標(biāo)NEFA、OCT以及TBIL同時(shí)升高可作為奶牛脂肪肝臨床診斷指標(biāo),LPS升高也可以作為奶牛發(fā)生脂肪肝的參考指標(biāo)。血漿生化指標(biāo),超聲檢查和體況評分可綜合診斷奶牛脂肪肝。2.成功構(gòu)建體外分離的犢牛肝臟線粒體氧化損傷模型,證實(shí)氧化損傷可導(dǎo)致線粒體內(nèi)OCT的活性下降。脂肪肝奶牛血清中OCT活性的上升其中一個(gè)可能機(jī)制是線粒體氧化應(yīng)激。
[Abstract]:Fatty liver of dairy cows is one of the nutritional and metabolic diseases, which is due to the disorder of liver lipid metabolism in dairy cows, resulting in the accumulation of excess lipids in the form of glycerol three fat (TG) in the form of liver, accompanied by severe liver dysfunction. The incidence of fatty liver in cows is over 50%, and cattle show significant decline in milk production, fecundity and immunity. As a result, the development of dairy cattle is seriously hindered. Therefore, it is very important to confirm the diagnosis of fatty liver in dairy cows in time. At present, the diagnosis of fatty liver can only be based on the liver biopsy, but the method has a high requirement on the operator's skills, which affects the production performance of the dairy cows, and makes the operation and feasibility of the production poor. In this study, the diagnosis of fatty liver in dairy cows was accurately diagnosed. This study explored the comprehensive diagnostic methods of fatty liver in cows and the possible diagnostic indicators. Furthermore, high fat could cause oxidative stress in the liver mitochondria of the cows, and the liver mitochondrial specific enzyme ornithine aminoformyl transferase (OCT) increased in the serum of fatty liver and mitochondria. The relationship between oxidative stress and the changes of mitochondrial OCT activity was not clear. In this study, the effects of mitochondrial oxidative damage on mitochondrial oxidative damage were studied in vitro. The effects of mitochondrial oxidative damage on the activity of OCT were investigated. The results are as follows: 1. using two mathematical statistical analysis methods using principal component analysis and cluster analysis, combined with clinical practice, 112 milk was used. AST, TBIL, TG, Ca, K+, K+, K+, NEFA, OCT and lipopolysaccharide (LPS) were analyzed in bovine plasma related to liver disease, and the clinical comprehensive diagnostic index of fatty liver in dairy cows was studied by ultrasonic examination and body condition score. The results showed that 112 dairy cows could be divided into three groups, 13 cows suspected of fatty liver, 84% of the postpartum cows, 7 in the healthy group and 92 in the latent group, and the incidence of 44%. plasma NEFA, OCT, TBIL, K+, AST and LPS in perinatal cows was lower. In the liver group, 10 cows showed bright spots in the liver and suspected calcified foci in the liver of 10 cows and suspected fatty liver. The mean value of the body condition score of the suspected fatty liver group was 4.25. The score was higher than the Fe2+/H2O2 system of Fenton reaction. The mitochondrial oxidative damage model of calf liver was constructed in vitro, and the activity of OCT in the mitochondria was detected. The effects of mitochondrial oxidative damage on the liver of calves showed that the content of mitochondrial malondialdehyde (MDA) and the swelling of mitochondria increased with the increase of H2O2 concentration, and the activity of mitochondrial succinic dehydrogenase (SDH) and OCT decreased in the mitochondria, and the concentration dependent relationship was found in a certain range. The free radical scavenger mannitol could significantly inhibit the OCT activity. Under the condition of Fenton reaction, the content of mitochondrial MDA and the degree of mitochondria swelling and the decrease of SDH activity in mitochondria. Under the condition of Fenton reaction, the mitochondrial oxidative damage model of calf liver was successfully constructed in vitro. In the high concentration Fe2+/H2O2 reaction system, the activity of OCT in the mitochondria was greatly reduced. We can draw the following conclusions: 1. the plasma biochemical indexes NEFA, OCT and TBIL can be used as the clinical diagnostic index of fatty liver in dairy cows. The elevation of LPS can also be used as a reference index of fatty liver in dairy cows. The plasma biochemical indexes, ultrasound examination and body condition score can be used to synthetically diagnose the calf liver.2. of milk fat fatty liver in vitro, and the calf liver line separated in vitro is successfully constructed. The oxidative damage model shows that oxidative damage can lead to the decrease of OCT activity in mitochondria. One of the possible mechanisms for the increase of OCT activity in the serum of fatty liver cows is mitochondrial oxidative stress.
【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S858.23
本文編號(hào)：2130865
[Abstract]:Fatty liver of dairy cows is one of the nutritional and metabolic diseases, which is due to the disorder of liver lipid metabolism in dairy cows, resulting in the accumulation of excess lipids in the form of glycerol three fat (TG) in the form of liver, accompanied by severe liver dysfunction. The incidence of fatty liver in cows is over 50%, and cattle show significant decline in milk production, fecundity and immunity. As a result, the development of dairy cattle is seriously hindered. Therefore, it is very important to confirm the diagnosis of fatty liver in dairy cows in time. At present, the diagnosis of fatty liver can only be based on the liver biopsy, but the method has a high requirement on the operator's skills, which affects the production performance of the dairy cows, and makes the operation and feasibility of the production poor. In this study, the diagnosis of fatty liver in dairy cows was accurately diagnosed. This study explored the comprehensive diagnostic methods of fatty liver in cows and the possible diagnostic indicators. Furthermore, high fat could cause oxidative stress in the liver mitochondria of the cows, and the liver mitochondrial specific enzyme ornithine aminoformyl transferase (OCT) increased in the serum of fatty liver and mitochondria. The relationship between oxidative stress and the changes of mitochondrial OCT activity was not clear. In this study, the effects of mitochondrial oxidative damage on mitochondrial oxidative damage were studied in vitro. The effects of mitochondrial oxidative damage on the activity of OCT were investigated. The results are as follows: 1. using two mathematical statistical analysis methods using principal component analysis and cluster analysis, combined with clinical practice, 112 milk was used. AST, TBIL, TG, Ca, K+, K+, K+, NEFA, OCT and lipopolysaccharide (LPS) were analyzed in bovine plasma related to liver disease, and the clinical comprehensive diagnostic index of fatty liver in dairy cows was studied by ultrasonic examination and body condition score. The results showed that 112 dairy cows could be divided into three groups, 13 cows suspected of fatty liver, 84% of the postpartum cows, 7 in the healthy group and 92 in the latent group, and the incidence of 44%. plasma NEFA, OCT, TBIL, K+, AST and LPS in perinatal cows was lower. In the liver group, 10 cows showed bright spots in the liver and suspected calcified foci in the liver of 10 cows and suspected fatty liver. The mean value of the body condition score of the suspected fatty liver group was 4.25. The score was higher than the Fe2+/H2O2 system of Fenton reaction. The mitochondrial oxidative damage model of calf liver was constructed in vitro, and the activity of OCT in the mitochondria was detected. The effects of mitochondrial oxidative damage on the liver of calves showed that the content of mitochondrial malondialdehyde (MDA) and the swelling of mitochondria increased with the increase of H2O2 concentration, and the activity of mitochondrial succinic dehydrogenase (SDH) and OCT decreased in the mitochondria, and the concentration dependent relationship was found in a certain range. The free radical scavenger mannitol could significantly inhibit the OCT activity. Under the condition of Fenton reaction, the content of mitochondrial MDA and the degree of mitochondria swelling and the decrease of SDH activity in mitochondria. Under the condition of Fenton reaction, the mitochondrial oxidative damage model of calf liver was successfully constructed in vitro. In the high concentration Fe2+/H2O2 reaction system, the activity of OCT in the mitochondria was greatly reduced. We can draw the following conclusions: 1. the plasma biochemical indexes NEFA, OCT and TBIL can be used as the clinical diagnostic index of fatty liver in dairy cows. The elevation of LPS can also be used as a reference index of fatty liver in dairy cows. The plasma biochemical indexes, ultrasound examination and body condition score can be used to synthetically diagnose the calf liver.2. of milk fat fatty liver in vitro, and the calf liver line separated in vitro is successfully constructed. The oxidative damage model shows that oxidative damage can lead to the decrease of OCT activity in mitochondria. One of the possible mechanisms for the increase of OCT activity in the serum of fatty liver cows is mitochondrial oxidative stress.
【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S858.23
【參考文獻(xiàn)】
相關(guān)期刊論文 前1條
1 周作榮,彭向欣;線粒體反應(yīng)氧體系與脂肪肝[J];中華肝臟病雜志;2003年02期
，本文編號(hào)：2130865
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