鹅细小病毒侵染雏鹅组织器官的氧化/脱氢酶及同工酶研究

酶是催化代谢途径的高效活性基因产物,应激环境和遗传基础的不同导致同工酶结构或活性差异,使不同组织和发育阶段的酶种类和活性表现特异性变化。采用生化分析试验技术和PAGE分析鹅细小病毒(GPV)感染雏鹅氧化/脱氢酶(LDH、ADH、POD、CAT)的活性及同工酶特征变异,结果表明,GPV感染雏鹅的脑、心脏、脾脏组织新出现1条POD同工酶谱带,肺脏组织消失1条同工酶谱带;GPV感染雏鹅的肝脏、脾脏组织分别缺失慢区3条和1条CAT同工酶谱带;GPV感染雏鹅组织器官的LDH和ADH同工酶仅显示1~2个慢区带,肺脏缺失1条慢区ADH同工酶谱带,ADH活性降低了13.61%~61.08%,心脏增高1.2倍;POD、CAT、LDH活性分别增强1.2~6、1.5~3.5、2~2.5倍,而脑、肺脏的CAT活性降低了40.29%和94.68%,心脏、肺脏的LDH活性降低了75.93%和91.81%。提示氧化/脱氢酶产生的广泛变异是GPV感染应激与宿主氧化/脱氢酶基因表达的互作效应,其酶的谱型、活性等生化特征变异,直接或间接调控代谢途径、影响生理机能及病理性能,敏感的氧化/脱氢酶是研究病毒基因型与宿主遗传易感性互作病理学机制的有效标记物。     

鹅细小病毒感染雏鹅血液的蛋白/酶活性及同工酶变异性研究

为了解鹅细小病毒(goose parvovirus,GPV)侵染的病理学致病机理,探究蛋白/酶的分子变异特征,本研究对GPV感染雏鹅血液中的蛋白质、保护酶、蛋白酶活性及其同工酶结构和功能等进行了生化—分子生物学分析.结果显示,GPV感染雏鹅的血液中(血浆和血细胞)蛋白酶活性和蛋白质含量、过氧化物酶(POD)、超氧化物歧化酶(SOD)及酯酶(Est)活性分别是对照组的1.46和1.63、1.51和1.49、1.50和1.14、1.36和1.73、1.30和1.53倍.GPV感染雏鹅的血浆中,SOD同工酶增加了2条谱带(134和239);Est同工酶增加2条快区主带,减少了2条慢区谱带;酶蛋白减少1条慢区谱带(304);蛋白质新增加4条主带(131、86、43、33 ku).而血细胞新出现2条POD同工酶谱带(93和203),分别增加了1条(160)中区SOD同工酶谱带、1个快区Est同工酶谱带、2个慢区酶蛋白谱(223和330)和4个蛋白主带(144、104、58、53 ku).提示GPV感染应激与寄主基因表达的蛋白质、保护酶、蛋白酶及同工酶相互网络协作会引起酶的谱型、活性等生化特征变异,直接或间接调控代谢途径与病理生化性能,增强机体对入侵GPV的防御应激性能.因此,蛋白质、保护酶、蛋白酶及同工酶是GPV感染的应激靶标,可作为雏鹅易感GPV致病机制研究的有效标记.     

Study on the Isozyme Variability and Activity of Protein/Enzymes from Gosling Blood Infected with Goose Parvovirus

In order to understand the pathogenesis of GPV (goose parvovirus),and explore the molecular variation of protein/enzyme,the protein,protective enzymes (Est,POD,SOD,protease) and isozyme from goslings blood infected with GPV were analyzed by biochemistry and molecular biology.The results showed that the activity of protease,content of protein,the activities of POD,SOD and Est (in plasma and blood corpuscle) from goslings infected with GPV were 1.46 and 1.63,1.51 and 1.49,1.50 and 1.14,1.36 and 1.73,1.30 and 1.53 times than that of control group,respectively.In the plasma of goslings infected with GPV,2 bands (134 and 239) of SOD isozyme appeared,2 fast-zone enzyme band appeared and 2 slow-zone enzyme band deleted in Est isozyme,1 slow-zone zymoprotein band (304) deleted and 4 new main protein bands (131,86,43 and 33 ku) appeared.In the blood corpuscle of goslings infected with GPV,2 new enzyme bands (93 and 203) of POD isozyme,1 medium-zone enzyme band (160) of SOD isozyme,1 fast-zone enzyme band of Est isozyme,2 slow-zone zymoprotein band (223 and 330) and 4 new main protein bands (144,104,58 and 53 ku) appeared.These results indicated that the interaction of GPV stress and host protein/enzymes and isozyme gene expression would result in the biochemical characteristics variation of activity and isozyme patterns of protein/enzymes,and directly or indirectly affect metabolic approach and pathological biochemical function on goslings infected with GPV,and enhance the defense and stress ability of GPV.Therefore,protein/enzymes and isozyme might be sensitive enzyme of GPV infection stress,and were effective marker of pathological mechanism on investigation of virus genotype interaction with genetic susceptibility of the host.     

淹水对紫花苜蓿南北方品种抗氧化酶和无氧呼吸酶的影响

以紫花苜蓿南方育成品种渝苜1号和北方地方品种新疆紫泥泉为材料,在幼苗长出4片真叶时进行淹水处理,在淹水的第10天测定植株的生物量,并分别在淹水的第0,2,4,6,8,10天测定叶片中的丙二醛(MDA)含量以及叶片抗氧化酶和根系无氧呼吸酶的活性,以探明两个品种对淹水胁迫的耐受性差异及其生理响应机制。结果表明,淹水胁迫使紫花苜蓿的生物量减少,但与新疆紫泥泉相比,渝苜1号生物量的降幅小。淹水时渝苜1号叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)等抗氧化酶活性比新疆紫泥泉的高,而叶片MDA含量比新疆紫泥泉的低;根系无氧呼吸酶活性在淹水胁迫中均增强,但渝苜1号的乙醇脱氢酶(ADH)和丙酮酸脱羧酶(PDC)活性增加明显,乙醇发酵更强;而新疆紫泥泉的乳酸脱氢酶(LDH)的活性增加更为突出,乳酸发酵强。淹水影响紫花苜蓿正常生长,但南方品种渝苜1号比北方品种新疆紫泥泉对淹水胁迫更具耐受性,因为前者具有比较高的抗氧化酶活性和比较强的乙醇发酵途径,有利于增强植株抗淹水胁迫能力。     

Some enzyme interrelationships in serum and organs of Egyptian camel

In 7 normal healthy Egyptian one-humped camels aged 3-4 years, the relationships were studied between enzyme activities of lactic dehydrogenase (LDH), creatine phosphokinase (CPK), alkaline phosphatase (ALP), acid phosphatase (ACP), and cholonesterase (CHE) of serum and organs as well as between ACP and ALP and between LDH and CPK. Liver, heart, kidney, and spleen tissue samples as well as serum were analysed for total enzyme activity. The following results were obtained: --The heart was the organ with the highest activities of ALP, LDH, and CPK, and it contained high values of CHE, whereas the lowest activities of all enzymes were recorded from serum. The spleen exhibited of the highest activity of ACP. --Each of the serum enzymes ALP, LDH, and CHE were in strong inverse relationship with the corresponding enzymes in the liver. Strong inverse relationships existed also between serum LDH and kidney LDH as well as between CPK in serum and heart. --Direct relationships were remarkable serum LDH and that of spleen and heart as well as between serum ACP and that of liver and heart. --Interrelationships were inverse between ACP and ALP in liver, kidney, and heart, but weak direct interrelationships were characteristic between the 2 enzymes in serum and spleen. --LDH was inversely interrelated with CPK in serum and heart.     

小鹅瘟病原学与检测方法的研究

从患病雏鹅的病料样本中分离鉴定出一株小鹅瘟野生强毒.采用鹅胚增殖病毒,以琼脂扩散试验从感染鹅胚尿囊液中检出小鹅瘟病毒抗原;以感染鹅胚尿囊液人工接种5日龄健康雏鹅,复制出与自然感染病例相类似的病变; 通过电镜观察到典型的小鹅瘟病毒粒子.该分离毒株核酸分子量为5 000 bp.制备的小鹅瘟沉淀抗原可以检测患病雏鹅血清中的抗体,监测治疗血清的滴度和疫苗接种的效果.标记的小鹅瘟荧光抗体能直接检测患病雏鹅组织中的病毒抗原.针对编码主要结构蛋白VP3的基因设计合成一对引物,扩增出与预期长度相符的特异性DNA片段.     

小鹅瘟病毒的分离鉴定与临床病变观察

从患病雏鹅的病料样本中分离鉴定出一株小鹅瘟野毒。采用鹅胚扩增病毒。以琼脂扩散试验从感染鹅胚尿囊液中检出小鹅瘟病毒抗原,用感染鹅胚尿囊液人工接种8日龄健康雏鹅,雏鹅表现出典型的小鹅瘟临床症状和剖检特征。     

肠炎沙门氏杆菌感染雏鸭脾脏过氧化氢酶及过氧化物酶活性的变化

通过肠炎沙门氏杆菌人工感染雏鸭,探讨肠炎沙门氏杆菌对禽类引发疾病的机理,测定了脾脏组织中POD及CAT活性。结果表明:POD活性较不稳定, 12h显著升高 (P<0.05), 36h、60h呈下降趋势并显著或极显著降低(P<0.05或P<0.01),84h后显著升高(P<0.05);CAT活性呈升高趋势,于 84h时显著高于健康对照组 (P<0.05);用氟苯尼考或中草药复方制剂治疗均可相应增强脾脏POD、CAT的活性。脾脏中CAT和POD参与了抵御肠炎沙门氏杆菌的过程。     

猪和绵羊组织器官乳酸脱氢酶同工酶酶谱比较

应用聚丙烯酰胺凝胶电泳法对猪和绵羊的心、肝、脾、肺、肾、胰、小肠７种组织器官及血清的乳酸脱氢酶同工酶酶谱进行比较研究，结果发现：①猪和绵羊各组织器官中ＬＤＨ同工酶共有５种区带，猪的ＬＤＨ３同工酶还可分为两条亚带；②猪和绵羊的肺、肾、小肠、胰、血清中ＬＤＨ区带数相同，其它组织器官各不相同；③猪和绵羊ＬＤＨ１同工酶的泳动速度最快，迁移率分别为５３．６４％、４１．８２％；ＬＤＨ５同工酶的泳动速度最慢，迁     

堆型艾美耳球虫感染后组织抗氧化功能的变化

通过测定堆型艾美耳球虫(Eimeria acervulina)感染鸡心脏、肝脏、脾脏、肺脏和肾脏组织中超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)和过氧化氢酶(CAT)的水平,研究E.acervulina感染对雏鸡的组织抗氧化功能的影响。结果表明,雏鸡在感染E.acervulina后,早期各组织SOD活性均低于对照组水平,但很快都又恢复到正常对照组水平;各组织中MDA含量均高于对照组水平;心脏、肝脏和脾脏中CAT活性在感染后下降,肺脏和肾脏中CAT活性在感染后升高;各组织GSH-Px活性在感染后均有不同程度的下降。表明病理氧化应激参与了球虫感染过程,损伤了机体的氧化平衡。     

三种鹿血清乳酸脱氢酶同功酶的电泳研究

采用聚丙烯酰胺凝胶电泳法对26头白唇鹿、41头马鹿和11头梅花鹿的血清乳酸脱氢酶（LDH）同功酶谱及其多态性进行了研究。结果发现：三种鹿的LDH都有LDH1、LDH2、LDH3和LDH5四种同功酶;LDH3同功酶具有1～3条亚带,LDH5具有显现酶活性和不显现酶活性两种表型而呈现多态性;白唇鹿的LDH同功酶酶谱及其多态性有较明显的种的特征。     

Tissue and plasma enzyme activities in juvenile green iguanas

OBJECTIVE: To determine activities of intracellular enzymes in 8 major organs in juvenile green iguanas and to compare tissue and plasma activities. ANIMALS: 6 green iguanas < 1 year old. PROCEDURE: Lysates of liver, kidney, epaxial muscle, heart, lung, spleen, small intestine, and pancreas were analyzed for alkaline phosphatase (ALP), lactate dehydrogenase (LDH), aspartate transaminase (AST), alanine transaminase (ALT), gamma-glutamyltransferase (GGT), creatine kinase (CK), glutamate dehydrogenase (GMD), and amylase (AMS) activities. RESULTS: In general, low tissue enzyme activity coincided with low plasma activity. The CK activity was high in epaxial muscle and the heart and low in all other tissues tested. The AMS activity was found exclusively in the pancreas. Moderate LDH and AST activities were found in all tissues. Low ALT and ALP activities were found in a variety of tissues. Plasma and tissue activities of GGT and GMD were low or undetectable. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that plasma CK activity may be muscle specific in iguanas, but high values may not always indicate overt muscle disease. The AMS activity may be specific for the pancreas, but the wide range of plasma activity would likely limit its diagnostic usefulness. Activities of AST and LDH may reflect tissue damage or inflammation, but probably do not reflect damage to specific tissues or organs.     

一株GPV的分离鉴定及致病性研究

为了确定导致吉林省某地区10日龄雏鹅发病的病原,从病死雏鹅的肝脏中分离到一株病毒,根据病毒的电镜照片,动物回归试验及PCR鉴定,确定为鹅细小病毒。PCR产物测序后经BLAST比对显示,分离毒株与NCBI收录的GPVVP3基因同源性均在92％以上,与GPV／CH／HLJ02／08VP3基因的同源性最高,达99％。致病性研究表明,分离株的ELD50为10^-6.5／0．2mL,毒力较强,且该毒株能被GPV标准血清所中和。     

Lactate dehydrogenase and creatine phosphokinase isoenzymes in tissues of laboratory animals

The lactate dehydrogenase (LDH) and creatine phosphokinase (CPK) isoenzyme distributions in tissues of the ICR mouse, Wistar rat, guinea pig and golden hamster were analyzed by histoelectrophoresis. Tissues obtained were as follows: liver, pancreas, stomach, small intestine, heart, femoral muscle, uterus, kidney, spleen, lymph node, cerebrum, spinal cord and erythrocyte. Histoelectrophoresis was for the direct analysis of LDH and CPK isoenzymes in the tissues and had high practical value compared with previous tissue-extraction methods. In tissues of the mouse, guinea pig and golden hamster, LDH isoenzymes showed five bands. In the rat, LDH isoenzyme was separated into four fractions. CPK isoenzyme showed three bands; BB, MB and MM. In some tissues, the MM band was separated into two sub fractions.     

Botulinum C3 enzyme changes the lactate dehydrogenase isozyme pattern of primary culture of neurons

Changes in the lactate dehydrogenase (LDH) isozyme pattern of primary culture of neurons treated with botulinum C3 enzyme were examined in order to elucidate the functional changes accompanying the morphological change that follows ADP-ribosylation of Rho protein. Primary neurons were prepared from the cerebrum of ICR mouse embryos on day 15. Neurons were cultured in MEM with 10% fetal calf serum at 37 degrees C. In the neurons treated with C3 enzyme, a typical morphological change was observed after 24 hr, and the LDH isozyme pattern was changed after 72 hr. The ratio of H-subunit to M-subunit in LDH was decreased by C3 treatment, suggesting the induction of a state of lower intracellular oxygen consumption in neurons in the primary cultures.     

外源NO及反向调控PEG胁迫下苜蓿萌发种子抗氧化酶及其同工酶动态的研究

以紫花苜蓿为材料,用一氧化氮供体硝普钠(SNP)及一氧化氮清除剂c-PTIO对苜蓿种子进行浸种处理,采用分光光度法和同工酶电泳技术来研究外源NO及反向调控对PEG胁迫下紫花苜蓿抗氧化酶活性及其同工酶的影响,并探讨NO调控苜蓿种子耐旱性的生理机制。结果表明:PEG胁迫下外施0.1 mmol·L^-1 SNP,第6天时较PEG处理POD活性降低了32.72%;第4天时SOD、CAT活性较PEG处理升高了10.48%、23.60%,有效缓解了PEG对紫花苜蓿萌发中种子的氧化损伤,提高其抗氧化能力。PEG胁迫下添加 c-PTIO抑制了苜蓿萌发期的抗氧化系统活性。从同工酶的谱带数量和强弱来看,POD同工酶各区带活力均随PEG胁迫时间的延长而增加,在第2天酶带只有1条,而第4天酶带呈现9条;SOD和CAT同工酶表达量变化不显著,但酶带强弱有一定变化,S3酶带随处理时间的延长表达量逐渐减弱;CAT同工酶谱带则一直保持2条带,无明显强弱变化。因此,外源NO在PEG胁迫下紫花苜蓿萌发中抗氧化酶快速响应并在维护氧自由基代谢平衡中起重要保护作用。     

细叶百合冷藏过程中鳞茎保护酶活性与休眠解除的关系

以细叶百合为试材,通过冷藏(5℃)解除鳞茎休眠,研究了细叶百合鳞茎解除休眠过程中鳞茎保护酶活性与糖代谢的变化规律。结果表明,鳞片、顶芽及鳞茎盘SOD活性在冷藏0~24 d内下降,除顶芽外,鳞茎各部位POD活性在0~24 d内呈下降趋势,外鳞片、顶芽及鳞茎盘中CAT活性在冷藏0~12 d内下降,冷藏中后期,SOD、CAT、ASP 活性升高,鳞片中POD的活性下降,顶芽及鳞茎盘POD、PPO在冷藏中期上升。各种代谢相关酶在不同器官中的作用并不完全相同。在低温处理36~60 d内,ASP、CAT活性快速上升,后期趋于稳定。SOD活性最低点出现在冷藏36~60 d,鳞茎各部位淀粉与CAT、PPO、ASP均表现为负相关性,SOD、POD、PAL与鳞片中淀粉表现为正相关性。36 d是鳞茎解除休眠的起点,60 d时鳞茎基本解除休眠。     

藏獒血液乳酸脱氢酶同工酶酶谱的电泳研究

采用聚丙烯酰胺凝胶电泳法对２１条藏獒的红细胞乳酸脱氢酶（ＲＢＣ－ＬＤＨ）和血清到脱氢酶（Ｓ－ＬＤＨ）同工酶酶谱及其多态性进行研究。结果发现：（１）ＲＢＣ－ＬＤＨ同工酶共有３条区带,呈现两种电泳表型,１１条藏契为ＬＤＨ２Ａ型（５７．８９％）,８条藏獒为ＬＤＨ２ａ（４２．１１％）,（２）Ｓ－ＬＤＨ共有５条区带,呈现两种电泳表型,２条藏獒为ＬＤＨ１Ａ（９．５２％）,１９条藏獒为ＬＤＨ１ａ（９０．４８％）     

桑天牛幼虫感染白僵菌后体内主要保护酶活性的变化

采用分光光度法测定了桑天牛幼虫被白僵菌感染后体内主要保护酶活性的变化。结果表明,桑天牛幼虫感染白僵菌后,为抵御白僵菌入侵,体内超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性在接种后初期迅速提高,但在接种后期均出现不同程度下降。其中,SOD、POD在接种后第2天达高峰,而CAT活性在第4天才达高峰,说明在桑天牛幼虫抵御白僵菌侵染的过程中,SOD和POD是首先发生作用的保护酶,CAT是继上述2种酶之后发生作用的酶系;并且POD活性始终高于对照组幼虫,是起主要作用的保护酶。     

Lactate dehydrogenase isoenzymes in the lungs of sheep with acute and chronic pneumonia

Total lactate dehydrogenase and the absolute and percentage levels of its isoenzymes were measured in lung lesions and macroscopically normal areas of lung from lambs with chronic proliferative exudative pneumonia and acute pasteurella pneumonia. Lung lesions had a higher total enzyme activity which was associated mainly with increases in the activity of the LDH4 and LDH5 isoenzymes, particularly in chronic pneumonia, and gave lung lesions a considerable potential for altering the serum isoenzyme distribution. Thus, the nature of any changes in the serum isoenzyme distribution will depend on whether the isoenzymes are released from abnormal or normal areas of lung. This appears to be the first report on lactate dehydrogenase isoenzymes in ovine pneumonia.