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《中国畜牧杂志》2017,(11)
实验旨在阐明牛酰基辅酶A氧化酶1(Acyl-Co A Oxidase 1,ACOX1)在肌内脂肪沉积过程中的作用,为优质肉牛品种培育提供理论依据。首先扩增牛ACOX1基因的CDS序列,构建ACOX1基因的过表达载体,然后将过表达载体转染3T3-L1细胞,利用油红O染色技术检测脂滴沉积情况,利用荧光定量PCR、Western blotting技术检测成脂分化基因PPARγ及C/EBPα的表达量。BLAST分析及酶切鉴定结果显示,成功构建了牛ACOX1基因的过表达载体;油红O结果表明,过表达ACOX1基因抑制了3T3-L1细胞的脂滴沉积;荧光定量PCR及Western blotting结果表明,过表达ACOX1基因显著性抑制了C/EBPα的表达量而对PPARγ的表达量无显著性影响。本研究结果显示,ACOX1可能抑制肌内脂肪沉积。 相似文献
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酚氧化酶在昆虫体液免疫反应中起着非常重要的作用。根据GenBank上登录的家蚕酚氧化酶原基因的cDNA序列设计特异引物,通过半定量BT-PCR技术检测了家蚕各组织酚氧化酶原基因的表达谱,发现该基因的转录表达具有组织特异性。我们的实验结果为进一步研究家蚕酚氧化酶原基因的功能提供了分子基础。 相似文献
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旨在探索过表达Fad24基因对猪DFAT细胞诱导成脂分化的影响。根据猪Fad24基因序列设计引物，克隆猪Fad24基因的全长CDS区片段，构建猪Fad24过表达载体，通过测序比对碱基和氨基酸序列，通过脂质体瞬时转染技术，将过表达Fad24载体转染猪DFAT细胞，从mRNA水平验证了Fad24基因在猪DFAT细胞中的过表达水平，然后将过表达Fad24的猪DFAT细胞在体外诱导成脂分化，从形态学、Fad24、成脂和成骨分化核心转录因子表达水平上进行了检测。结果表明，猪Fad24过表达载体碱基序列一致性高达99.8%,氨基酸序列一致性高达100%;过表达Fad24的猪DFAT细胞Fad24的表达量高出正常猪DFAT细胞表达量约45倍；成脂分化诱导5 d时，与对照组的正常分化比较，过表达Fad24的猪DFAT细胞成脂分化完全被抑制，相反强力转向成骨分化，细胞积聚成许多小结节，诱导7 d时已形成典型的成骨样大结节，表面可见细胞分泌物；经茜素红染色鉴定，证实细胞分泌物中含有矿化物质沉淀；经Real-time PCR检测，证实过表达Fad24试验组显著下调成脂分化核心转录因子PPARγ2、SREBP... 相似文献
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脂联素是一种由脂肪组织分泌的具有多种生物学功能的特殊蛋白,在增强脂肪酸氧化、抗炎症反应、抗糖尿病等方面起重要作用。脂联素通过AdipoR1和AdipoR2这两种受体的介导经过AMPK、PPAR、p38MAPK等信号通路来发挥生物学作用。脂联素及其受体AdipoR1和AdipoR2能在多种组织器官中表达,AdipoR1主要在肌肉组织中表达,AdipoR2则高表达于肝脏组织。此外,脂联素及其受体还能在下丘脑、垂体、子宫、胚胎等多种生殖腺和生殖组织中表达,说明脂联素在调控动物生殖及胚胎生长发育方面起重要作用。 相似文献
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《中国畜牧兽医文摘》2016,(8)
通过对兔血清中超氧化物歧化酶(SOD)和谷胱苷肽(G-SH)活性的影响研究。表明杜仲茶能显著提高兔血清中超氧化物歧化酶(SOD)和增加谷胱苷肽(G-SH)活性,对防止衰老、预防和缓解老年疾病症状具有作用。这对杜仲资源的开发有着重要意义! 相似文献
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口蹄疫病毒VP1-3免疫表位基因与猪γ-干扰素基因的融合表达 总被引：4，自引：0，他引：4
将人工合成的口蹄疫病毒VP1-3上的抗原表位基因和猪γ-干扰素基因串联入原核表达载体pGEX-KG中,经酶切鉴定及测序表明重组载体中二者以接头融合的形式构建成重组表达质粒。将该质粒转化BL21后经IPTG诱导,实现了重组融合蛋白的高效表达,表达产物经SDS-PAGE和Western blot分析,重组蛋白分子量约为66 Ku,与预期大小相符。薄层扫描分析显示表达的重组蛋白占菌体总蛋白的37.4%,且主要以包涵体的形式存在。包涵体以SKL变性后,经PEG4000、氧化型谷胱苷肽和还原型谷胱苷肽复性。粗提产物以CPE_(50)测得重组蛋白在MDBK细胞上的抗水泡性口炎病毒(Vesicular stomatitis virus,VSV)的活性达到1600 U/mL,说明表达产物具有干扰素的生物学活性,为下一步基因工程疫苗的研究奠定了基础。 相似文献
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体外原代培养犊牛肝细胞,添加0、16、64和128μg/L脂联素（adiponectin,ADPN）,分别培养4、8h后,提取细胞总RNA。应用实时荧光定量PCR方法,检测脂氧化关键酶脂酰辅酶A氧化酶（ACO）、肝脏脂肪酸结合蛋白（L-FABP）、肉碱脂酰转移酶-Ⅰ（CPTⅠ）、肉碱脂酰转移酶-Ⅱ（CPTⅡ）mRNA表达变化。结果显示,在ADPN作用4h后,ACO、L-FABP、CPTⅠ、CPTⅡ基因mRNA水平均随ADPN浓度的增加而增加,且均显著高于对照组,呈剂量依赖关系;作用8h时,ACO表达也增加,但差异不显著,L-FABP在中高剂量组显著高于对照组,CPTⅠ和CPTⅡ在各ADPN处理组均显著高于对照组。结果表明,ADPN可显著促进脂氧化关键酶的表达,增强肝脏脂氧化作用,减少肝脂储存。 相似文献
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过表达FGF10促进山羊皮下前体脂肪细胞分化 总被引：2，自引：2，他引：0
本研究在构建山羊FGF10过表达腺病毒载体的基础上,阐明过表达FGF10基因对山羊皮下前体脂肪细胞分化的影响及可能的作用机制。本试验利用胶原酶消化法分离获得山羊皮下前体脂肪细胞,采用AdEasy腺病毒包装系统成功构建过表达腺病毒pAdTrack-CMV-FGF10,并感染细胞。采用油红O染色方法从形态学上观察FGF10对成脂分化的影响;利用qPCR技术检测脂肪细胞分化标志基因、脂代谢相关基因、成纤维细胞生长因子受体(fibroblast growth factor receptors,FGFRs)和Kruppel样因子家族(Kruppel like factors,KLFs)mRNA的相对表达变化。结果显示,在过表达FGF10后的第2天,皮下脂肪细胞中脂滴聚集显著多于对照组,C/EBPα、LPL、ACACA、FGFR1和FGFR3的相对表达水平极显著上调(P0.01),FASN和ATGL的相对表达水平分别出现显著上调(P0.05)和显著下调(P0.05),同时,过表达FGF10显著上调KLF家族成员KLF8-10、16和17基因mRNA相对表达水平(P0.05),显著下调KLF1、2、4和15基因的相对表达水平(P0.05)。结果表明,过表达FGF10基因可能通过调控C/EBPα、LPL、FASN、ACACA、ATGL和KLFs部分成员的表达促进山羊皮下前体脂肪细胞的分化及脂滴聚集,结果为进一步阐明其调控山羊不同部位脂肪沉积的分子机理提供重要的数据支撑。 相似文献
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Xiaoling Chen Lu Xiang Gang Jia Guangmang Liu Hua Zhao Zhiqing Huang 《Animal Science Journal》2019,90(8):990-998
The study was conducted to investigate the effects of dietary leucine on antioxidant activity and expression of antioxidant‐ and mitochondrial‐related genes in longissimus dorsi muscle and liver of piglets. Three diets were formulated with different levels of supplemented leucine (0%, 0.25%, 0.5%). Results showed that supplementation of 0.25% leucine significantly increased antisuperoxide anion (ASA) and antihydroxyl radical (AHR) levels and activities of total superoxide dismutade (T‐SOD), glutathione peroxidase (GPx), glutathione S‐transferase (GST), and total antioxidant capacity (T‐AOC) in serum, longissimus dorsi muscle and liver of piglets as compared with the control group. The SOD2, catalase (CAT), GPx, GST, glutathione reductase (GR), and nuclear factor erythroid 2‐related factor 2 (Nrf2) mRNA levels in longissimus dorsi muscle and liver were significantly increased by 0.25% leucine supplementation. However, the malondialdehyde (MDA) content and the mRNA level of Kelch‐like ECH‐associated protein 1 (Keap1) exhibited an opposite tendency. Additionally, supplementation of 0.25% leucine significantly increased the mRNA levels of mitochondrial‐related genes in longissimus dorsi muscle and liver of piglets. Results suggested that supplementation of 0.25% leucine improved antioxidant activity and mitochondrial biogenesis and function of piglets, which was related to the increase in antioxidant enzymes activities and upregulation of expression of antioxidant‐ and mitochondrial‐related genes. 相似文献
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Changes in the regulation and activity of glutathione redox system,and lipid peroxidation processes in short‐term aflatoxin B1 exposure in liver of laying hens 
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下载免费PDF全文 M. Erdélyi K. Balogh C. Pelyhe B. Kövesi M. Nakade E. Zándoki M. Mézes B. Kovács 《Journal of animal physiology and animal nutrition》2018,102(4):947-952
The purpose of this study was to investigate the short‐term (48 hr) effects of feeding aflatoxin contaminated diet (170.3 μg/kg AFB1) in 49‐week‐old laying hens. Liver samples were taken at 12‐hr intervals. Feed intake, body weight, absolute and relative liver weight were the same in groups. However, there was no feed intake during both dark periods (between 12nd to 24th and 36th to 48th hours of the experiment); therefore, aflatoxin intake was also negligible. Markers of initial phase of lipid peroxidation, conjugated dienes and trienes did not change as effect of aflatoxin, but terminal marker, malondialdehyde content was significantly higher at 12 hr as effect of aflatoxin. No significant difference was found in reduced glutathione concentration and glutathione peroxidase activity between the groups. Expression of glutathione peroxidase 4 gene (GPX4) was significantly reduced due to aflatoxin treatment at 12 and 24 hr, but induced later, while glutathione reductase gene (GSR) expression was significantly lower at 24 hr and glutathione synthetase gene (GSS) in aflatoxin‐treated group at 12 hr. The results suggest that aflatoxin induced oxygen‐free radical formation, but it did not reach critical level during this short period of time to cause activation of the expression of glutathione system. 相似文献
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Serum vitamin E and blood glutathione peroxidase values of horses with degenerative myeloencephalopathy 总被引：1，自引：0，他引：1
S G Dill F A Kallfelz A deLahunta C H Waldron 《American journal of veterinary research》1989,50(1):166-168
Serum vitamin E and blood glutathione peroxidase values were determined in 40 horses with a histologically confirmed diagnosis of degenerative myeloencephalopathy and in 49 age-matched control horses with normal neurologic function. Significant differences were not detected in serum vitamin E or blood glutathione peroxidase values between horses affected with degenerative myeloencephalopathy and control horses. These findings fail to support a reported role of vitamin E deficiency as a cause of equine degenerative myeloencephalopathy. 相似文献
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本试验旨在研究铁过量或缺乏对新生仔猪血清生化指标及肝脏hepcidin mRNA表达量的影响.挑选新出生的“杜长大”三元杂交仔猪15头[平均体重为(1.22±0.13)kg],随机分为3组,即缺铁组、正常组和铁过量组,每组5个重复,每个重复1头猪.3和7日龄时,缺铁组分别注射1 mL生理盐水,正常组分别注射1 mL右旋糖酐铁(含铁150 mg),铁过量组分别注射3 mL右旋糖酐铁(含铁450 mg).7日龄时,将所有仔猪全部处死,采集血清,并分离肝脏和脾脏,以测定血清生化指标、机体铁含量和肝脏hepcidin mRNA表达量.结果表明:肝脏、脾脏和血清中铁的含量均随着注射铁量的增加而显著或极显著增加(P ＜0.05或P＜0.01).与正常组相比,铁过量组血清中血红蛋白、球蛋白、总蛋白、丙二醛含量以及谷胱甘肽过氧化物酶、过氧化物酶活性显著或极显著升高(P＜0.05或P＜0.01),超氧化物歧化酶活性显著降低(P＜0.05);而缺铁组血清中血红蛋白、球蛋白、总蛋白、丙二醛含量以及谷胱甘肽过氧化物酶、过氧化氢酶、过氧化物酶活性则显著或极显著降低(P＜0.05或P＜0.01),超氧化物歧化酶活性显著升高(P＜0.05).与正常组相比,铁过量组仔猪肝脏中hepcidin mRNA表达量极显著升高(P＜0.01),而缺铁组则极显著降低(P＜0.01).由此得出,铁过量或缺乏均会影响新生仔猪机体的免疫功能和抗氧化功能;铁过量可提高新生仔猪机体铁含量和肝脏中hepcidin mRNA表达量,铁缺乏则会降低新生仔猪机体铁含量和肝脏中hepcidin mRNA表达量. 相似文献
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将pGEX-3X中由血吸虫（Schistosomajaponicum）编码的26kD谷胱甘肽转移酶基因Sj26和凝血因子Xa凝血酶位点引入到BacPAK8构建了融合表达杆状病毒转移载体BacSj26;将28kD的谷胱甘肽转移酶基因通过PCR突变消除终止密码后,引入BacPAK8构建了融合表达转移载体BacGST28。将P(MFHT)中的6×His序列克隆进BacPAK8,从而构建了6×His融合表达转移载体BacHis。这些融合表达转移载体的构建,为表达产物的一步纯化奠定了基础。 相似文献
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谷胱甘肽硫转移酶(GSTs)是一种解毒酶。为明确柞蚕谷胱甘肽硫转移酶-theta(GSTT)在柞蚕丝素蛋白准备和合成过程中所起的作用,采用生物信息学的方法克隆了柞蚕GSTT基因,并分析其在5龄幼虫丝腺中的表达规律。根据家蚕GSTT基因序列设计引物,获得了柞蚕GSTT基因的cDNA序列(651 bp,编码216个氨基酸,Gen-Bank登录号:EU541490)。该基因编码的蛋白与家蚕GSTT的同源性为89%。利用柞蚕18S核糖体蛋白基因作为内参,对柞蚕GSTT在不同时期的表达情况进行实时定量PCR检测,结果发现柞蚕GSTT的mRNA表达量在5龄第4天达到最高,后期逐渐下降。研究表明,在柞蚕丝素蛋白合成的5龄期,GSTT表达量大量提高,推测其主要功能为帮助柞蚕排除体内过多的氨基酸,以达到排毒目的。 相似文献
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A sequential biopsy sampling method was used to investigate oxidant and antioxidant gene responses in resistant sheep challenged with Haemonchus contortus larvae or a sham saline challenge. The expression of key sheep oxidant and antioxidant producing genes were measured in sequential samples removed from the abomasums at days 0, 1, 2, 3, 5, 7 and 28 post challenge. Gene expression levels at each time point were compared to expression at day 0, and levels of the various genes were also correlated to other markers of infection including immune cell counts and cytokine gene expression. The early response to larval challenge infection in resistant animals was marked by a divergence of two groups of host oxidant producing genes: the dual oxidase group (DUOX2/DUOXA2) showing increases in expression to day 7, while members of the phagocytic NADPH oxidase (PHOX) group showed significant decreases in expression. The change in DUOX2 expression between days zero and seven, when host resistance to infection is mediated, was negatively correlated to final worm burden suggesting NADPH oxidase expression may play a role in parasite expulsion. Expression of the DUOX group oxidants was positively correlated to expression of the Th2 cytokine IL4. Changes in host antioxidant pathways between different members of the glutathione peroxidase family (intestinal and plasma GPX) and genes involved in glutathione metabolism were also observed. This first study of the putative roles of oxidant production by the dual oxidase group, antioxidant glutathione pathways, immune cell populations, and cytokine profiles, in the development of resistance to infection by hyperimmune sheep are discussed. 相似文献
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Julia B. Montgomery Jeffrey J. Wichtel Maureen G. Wichtel Mary A. McNiven J T. McClure Fred Markham David W. Horohov 《Canadian journal of veterinary research》2012,76(4):281-291
The effects of selenium (Se) supplementation and source on equine immune function have not been extensively studied. This study examined the effects of oral Se supplementation and Se source on aspects of innate and adaptive immunity in horses. Fifteen horses were assigned to 1 of 3 groups (5 horses/group): control, inorganic Se (sodium selenite), organic Se (Se yeast). Immune function tests performed included: lymphocyte proliferation in response to mitogen concanavalin A, neutrophil phagocytosis, antibody production after rabies vaccination, relative cytokine gene expression in stimulated lymphocytes [interferon gamma (IFNγ), interleukin (IL)-2, IL-5, IL-10, tumor necrosis factor alpha (TNFα)], and neutrophils (IL-1, IL-6, IL-8, IL-12, TNFα). Plasma, red blood cell Se, and blood glutathione peroxidase activity were measured. Plasma and red blood cell Se were highest in horses in the organic Se group, compared with that of inorganic Se or control groups. Organic Se supplementation increased the relative lymphocyte expression of IL-5, compared with inorganic Se or no Se. Selenium supplementation increased relative neutrophil expression of IL-1 and IL-8. Other measures of immune function were unaffected. Dietary Se content and source appear to influence immune function in horses, including alterations in lymphocyte expression of IL-5, and neutrophil expression of IL-1 and IL-8. 相似文献
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谷胱甘肽和活性氧对哺乳动物配子作用的研究进展 总被引：1，自引：0，他引：1
文章综述了近年来关于谷胱甘肽对哺乳动物精子、卵母细胞和胚胎早期发育的作用及活性氧对精子功能的影响。在雄性和雌性配子中,谷胱甘肽起着保护精子和卵母细胞免受氧化损害的作用。在卵母细胞减数分裂中谷胱甘肽具有维持纺锤体形态的作用,受精后,巯基在精核的形成中起着积极的作用,并促进早期胚胎发育到囊胚阶段。在卵母细胞成熟过程中,谷胱甘肽的浓度发生着变化。它的合成受促性腺激素的调节,在精子的发生过程中随着精子的成熟,谷胱甘肽的浓度逐渐降低,但活性氧对精子功能也有多方面的作用。文章对卵丘细胞中小分子巯基化合物在谷胱甘肽合成中的重要作用也进行了综述。 相似文献