类维生素辅酶Q10功能与应用研究进展

辅酶Q10具有参与呼吸链电子传递、细胞抗氧化、阻止细胞凋亡、增强免疫等功能。辅酶Q10也是优良的抗氧化剂,对肉鸡腹水症及猝死症有明显的防控作用。     

辅酶Q10的制备及应用新进展

辅酶Q10是一种广泛存在于生物体中的重要生理活性物质,具有提高人体免疫力、增强抗氧化能力、延缓衰老等功能.目前,生物合成法是工业生产制备辅酶Q10的主要方式和未来趋势,辅酶Q10的应用正从医药领域逐渐拓展到食品、保健品和化妆品等更宽广的领域.对辅酶Q10的制备方法及应用新进展进行了概述.     

辅酶Q10对腹水症敏感肉鸡肝线粒体功能及抗氧化能力的影响

本文旨在研究添加辅酶Q10对腹水症敏感肉鸡肝脏线粒体功能及抗氧化能力的影响。选用1日龄艾维茵肉公鸡160只,随机分为4组,分别为大肠杆菌内毒素注射(lipopolysacchride,LPS)组(1.0 mg/kg体重剂量,腹腔注射)、LPS注射 40 mg/kg辅酶Q10添加组、生理盐水注射组、生理盐水 40 mg/kg辅酶Q10添加组,每组40只,设5个重复,每个重复8只。肉鸡饲以全植物性玉米-豆粕基础日粮,试验期6周。辅酶Q10从1日龄开始添加,10日龄开始突然关闭水暖,进行低温处理(12~15℃)以提高肉鸡腹水症敏感性。试验结果显示:(1)添加辅酶Q10肉鸡红细胞压积(PCV)、腹水心脏指数(AHI)显著降低(P<0.05)。(2)添加辅酶Q10肉鸡肝脏线粒体状态3、状态4呼吸速率、呼吸控制率(RCR)、磷氧比(P/O)变化虽不显著(P>0.05),但线粒体呼吸链复合体Ⅱ Ⅲ的质子转移与电子传递关系(H /2e)比值显著提高(P<0.05)。(3)添加辅酶Q10细胞色素C氧化酶(CCO)和H -ATP酶活性提高(P<0.05),但对线粒体NADH-细胞色素C还原酶(NCCR)和琥珀酸-细胞色素C还原酶(SCCR)活性都没有显著影响(P>0.05)。(4)添加辅酶Q10时,线粒体抗活性氧能力(ARC)显著提高(P<0.05),脂质过氧化产物丙二醛(MDA)含量显著降低(P<0.05),血清中总超氧化物歧化酶(T-SOD)活性显著升高(P<0.05),而线粒体内T-SOD、血清中过氧化氢酶(CAT)、抗超氧阴离子能力(ASAC)、ARC没有显著变化(P>0.05)。本试验结果表明,添加辅酶Q10能够在某种程度上改善腹水症敏感肉鸡的肝脏线粒体功能以及一些抗氧化能力。     

辅酶Q_(10)对鸡代谢调控作用及其机理

本文综述了辅酶Q10在鸡代谢调控中的作用,主要包括辅酶Q10对肉仔鸡腹水综合症、胆固醇代谢以及蛋鸡代谢的影响;并阐述了辅酶Q10对其代谢调控作用的可能机理。     

辅酶Q10对肉仔鸡生产性能的影响以及对腹水症的敏感性研究

180只AA雄性肉仔鸡随机分为3组,每组6个重复,8日龄起分别饲喂代谢能为13.2MJ/kg的日粮,辅酶Q10添加量分别为0、20、40mg/kg.15～21日龄关闭水暖进行较低温处理(15～18℃)以诱发腹水症,并研究辅酶Q10添加对肉仔鸡生产性能的影响以及对腹水症的敏感性.结果表明在本试验条件下,肉仔鸡日粮中添加不同水平的辅酶Q10对生产性能的影响不显著,而能显著降低其对腹水症的敏感性.日粮添加辅酶Q10能够显著降低肉仔鸡腹水症的发病率和死亡率,降低肉仔鸡红细胞渗透脆性(EOF)(P≤0.05).虽然辅酶Q10添加与否对血液红细胞压积(PCV)没有显著影响,但40mg/kg的辅酶Q10添加量同20mg/kg的添加量相比,PCV值显著降低(P≤0.05),除了36日龄时添加40mg/kg辅酶Q10显著降低了肉仔鸡的肺动脉舒张压(PADP)外,各试验组肉仔鸡右心室内压(RVP)、肺动脉压(PAP)、右心室内压最大变化速率(±dp/dtmax)等均没有表现出显著变化,而40mg/kg的辅酶Q10组显著降低了腹水心脏指数(AHI)(P≤0.05).辅酶Q10的这种有益作用可能与其为心肌细胞充足供能、降低血液的红细胞渗透脆性有关.     

辅酶Q10对湖羊精液常温保存效果的影响

本实验旨在探究辅酶Q10对湖羊精液常温保存效果的影响.选取3只1～2岁的健康湖羊采集精液,在基础稀释液中添加0、5、50、250、500μmol/L的辅酶Q10,检测在20℃保存过程中的湖羊精子活力、活率、曲线速度、质膜完整率、顶体完整率和活性氧(ROS)含量等指标.结果表明:随着在常温保存稀释液中添加辅酶Q10浓度的...     

大豆异黄酮和辅酶Q10联合添加对高能饲料引起的肉鸡腹水和猝死的影响

试验旨在研究大豆异黄酮和辅酶Q联合添加对肉鸡腹水和猝死发生的影响。选用1日龄AA肉公鸡,采用3×3因子设计,随机分为9个处理,每个处理10个重复。1～7日龄饲喂基础日粮,8日龄起饲喂添加大豆异黄酮和辅酶Q试验日粮。大豆异黄酮添加水平分别为0、10、20 mg/kg饲料,辅酶Q添加水平分别为0、20、40 mg/kg饲料。结果表明:10 mg/kg大豆异黄酮+40 mg/kg辅酶或20 mg/kg大豆异黄酮+20 mg/kg辅酶能显著减少心包积液、抑制肉鸡右心肥大的发生、降低红细胞压积、增加颈动脉血氧分压(P<0.05),降低了肉鸡腹水和猝死的发生率,证实了大豆异黄酮和辅酶Q抑制了腹水和猝死的发生发展过程。     

辅酶Q10注射液热原标准的研究

目的 :探讨细菌内毒素检查法测定辅酶Q10注射液中细菌内毒素含量的可行性,并以此建立辅酶Q10注射液热原的质量标准。方法:按照《中国药典》2010版二部XIE进行。结果:细菌内毒素法能够用于辅酶Q10注射液的细菌内毒素的测定,检测结果与家兔法一致,其质量内控指标为:将样品稀释400倍后,按《中国药典》2010版二部细菌内毒素法测定,应符合规定。结论:细菌内毒素法质量内控指标可用于辅酶Q10注射液生产中的质量控制。     

辅酶Q_(10)对腹水症肉鸡脂质代谢以及抗氧化能力的影响

研究日粮中添加辅酶Q10对腹水症肉鸡脂质代谢以及抗氧化能力的影响。将180只AA雄性肉仔鸡随机分为3组,每组6个重复,8日龄起在日粮中添加不同水平的辅酶Q10,分别为0、20、40mg/kg。15-21日龄关闭水暖进行较低温处理(15-18℃)以诱发腹水症。结果表明:日粮中添加40 mg/kg辅酶Q10显著降低腹水症肉鸡机体内TG、TC和LDL-C含量(P<0.05),而HDL-C没有变化;显著提高TAOC和降低MDA含量(P<0.05)。这表明辅酶Q10相对长期添加时能够改善肉鸡脂质代谢以及总抗氧化能力。     

辅酶Q10对绒山羊精液冷冻保存效果的影响

旨在探讨辅酶Q10对绒山羊精液冷冻保存效果的影响。利用添加不同浓度辅酶Q10(4、40、400?滋g/mL)的精液冷冻稀释液对绒山羊精液样本进行冷冻保存,待冷冻精液解冻后,采用流式细胞仪和计算机辅助精液分析系统(CASAS)分别检测不同精液样本的精子活率、质膜完整率、顶体完整率、DNA完整率、线粒体膜电位和细胞内ROS水平。结果表明,当冷冻稀释液中添加浓度为40μg/mL辅酶Q10时,经历冷冻—解冻过程的绒山羊精液样本的精子活率、质膜完整率、顶体完整率均显著高于对照组(P<0.05);在冷冻稀释液中添加浓度为40μg/mL或400μg/mL的辅酶Q10均能显著提高线粒体膜电位并降低细胞内ROS水平(P<0.05)。综上所述,在冷冻稀释液中添加40μg/mL的辅酶Q10能够显著提高绒山羊精子抗氧化能力和冷冻保存效果。     

Homeopathic prophylaxis in dairy cows on an organic farm part 1--fertility

The objective of the study was to assess the efficacy of different prophylactically applied homeopathic compounds on health and fertility during the periparturient period on an organic dairy farm. In a randomised double blinded study 146 dairy cows were enrolled in two treatment groups.The average milk yield was about 5100 kg per cow per lactation. The treatment group received the homeopathic compounds Carduus comp. and Coenzyme comp. at drying off, Traumeel on the day of calving, Lachesis comp. on day 7 post partum (p.p.) and Carduus comp. and Coenzyme comp. on day 14 days p.p. The control group followed the same protocol with a placebo (physiological saline solution). Each drug was administered subcutaneously in a dosage of 5 ml. At drying off, the day of calving and in weekly intervals until day 35 p.p. clinical examinations as well as blood sampling were performed. The effect of treatment was measured by clinical parameters, reproductive performance and serum profiles (Ca, P, AST, Urea, Bilirubin). Data of reproductive performance (days to first service, days open, conception rate) were compared between treatment groups and to those in the previous lactation. There was no significant difference between both treatment groups. Cows of the treatment group had an earlier onset of cyclic activity, especially when milk yield was considered as an influencing factor (82% vs. 57%, P < 0,05). In contrast the cows of the treatment group had a significant lower submission rate. The prophylactic treatment of all cows did not have an effect in general, but in cows with increased milk yield, especially in the current lactation. The reproductive performance in the previous lactation did not have any effects on the success of the homeopathic treatment. Reproductive performance in the herd could be enhanced slightly compared to the previous lactation.     

Use of CD10 as a marker of canine mammary myoepithelial cells

CD10 is an important cell marker in the diagnosis of acute lymphoblastic leukaemia and of breast myoepithelial (ME) cells in humans. The objective of this study was to assess the value of CD10 as a marker of canine ME cells using immunohistochemistry on routinely processed normal, dysplastic and neoplastic mammary tissue. Five different CD10 positive cell types were identified on the basis of cell morphology, pattern of immunoreactivity, and on the co-expression of additional cell lineage-specific markers.Type 1 cells were typical fusiform cells with a ME cell phenotype (calponin- and cytokeratin [CK] 14-positive, CK8/18-negative). Type 2 cells were typical or atypical polyhedral cells with a luminal epithelial (LE) cell phenotype (calponin- and CK14-negative, CK8/18-positive). Type 3 cells had a type 1 phenotype with variable morphology, and type 4 were atypical neoplastic cells with a mixed ME/LE phenotype. Type 5 cells were typical fusiform cells with a stromal phenotype.Type 1 cells were considered normal ME cells and were found in all sample types; type 2 cells were considered normal or neoplastic LE cells and were also found in all sample types; types 3 and 4 cells were restricted to tumour samples and to malignant tumours, respectively, and type 5 cells were found in all sample types, although predominantly in neoplastic tissue. The findings indicate that the CD10 antigen is a sensitive (although not specific) marker of canine ME cells in normal, dysplastic and neoplastic mammary tissue. Differences in the distribution and staining intensity of CD10-positive cells suggest a number of potential roles for this protein in the pathogenesis of canine mammary neoplasia.     

猪瘟免疫猪接种PRRSV nsp2Δ1882-2241弱毒疫苗后IL-12、IL-10、TNF-α应答特征

不同时间段采集猪的血液样本,利用ELISA方法检测血清和细胞培养上清中IL-12、IL-10、TNF-α等3种细胞因子的水平,分析猪双重免疫CSFV、PRRSV弱毒苗后血清和细胞培养上清中细胞因子的变化规律。结果显示,IL-12在血清中的浓度显著高于细胞培养上清中的浓度(P<0.01),在CSFV高抗体PRRSV高抗体组中28d采血的细胞培养上清中IL-12的浓度高于14d采血的细胞培养上清中的浓度(P<0.05);IL-10血清中水平显著低于细胞培养上清中水平(P<0.01),在CSFV高抗体PRRSV高抗体组中28d采血的细胞培养上清中的IL-10浓度显著低于14d采血的细胞培养上清中的浓度(P<0.01);TNF-α在血清中水平显著低于细胞培养上清中水平(P<0.01),在3组中28d采血的细胞培养上清液中TNF-α浓度高于14d采血的细胞培养上清液中浓度(P<0.05)。结果表明,2种疫苗特异性抗体应答高时,IL-12水平显著上调,IL-10水平显著下调,提示上调IL-12及下调IL-10都是基因缺失疫苗发挥效力的潜在机制。TNF-α总体水平上升,但在血清中水平较低。     

Multifocal cerebellar granular layer necrosis in traumatically head-injured lambs.

In a blunt, nonmissile, head impact model of traumatic brain injury in 4-5-week-old Merino lambs, multiple foci of internal granular layer necrosis were found in all 10 impacted animals. This lesion has not previously been reported after human or animal head injury. Temporal lobe impact contusions, predominantly microscopic (8/10) and contralateral contusions (2/10), parenchymal (10/10) and subarachnoid (10/10) hemorrhage, and widely distributed axonal injury were also observed. Although the precise pathogenesis of this focal granule cell necrosis and often attendant red cell change in Purkinje cells was unclear, an ischemic etiology due to trauma-related vascular damage is postulated.     

成年小鼠雄性生殖细胞的冷冻保存

在含10％小牛血清（NBS）的DMEM培养基中，分别各添加5％，10％，15％，20％和25％的二甲基亚砜（DMSO）,丙二醇（PG），乙二醇（EG）和甘油（G），对成年小鼠睾丸生殖细胞冷冻保存；复苏后台盼蓝染色测定细胞复苏率。结果显示，5％－25％DMSO冻存液组的细胞复苏率分别为88.5%,88.0%,65.6%及51.3%;5%-25%PG冻存液组的细胞复苏率分别为87.2%,86.4%,79.0%,73.4%及40.1%.;5%-25%EG冻存液组的细胞复苏率分别为6.6%,80.9%,60.8%,51.3%及30.0%;5%-25%G冻存液组的细胞复苏率分别为86．5％，86．3％，65．3％，36．0％及31．4％。其中各抗冻剂5％和10％组的细胞复苏率最高，与15％组相比均存在显著或极显著差异。4种抗冻剂的最高细胞复苏率之间无显著差异。DMSO，PG，EG和G分别冷冻保存成年小鼠睾丸生殖细胞对的最小损失率分别为4．8％，6．1％，6．7％，6．8％。结果表明，采用慢速冷冻时，DMSO，PG，EG及G均适宜用作成年小鼠睾丸生殖细胞的抗冷冻剂，最佳使用含量均为5％－10％。成年小鼠睾丸生殖细胞分别在含5％－10％的DMSO，PG，EG和G的DMEM（含10％NBS）冻存液中，2步慢速降温，液氮储存，37℃水浴复苏，是一种具有较高复苏率的冷冻保存方法。     

猪瘟免疫猪接种PRRSVnsp2A1882-2241弱毒疫苗后IL-12、IL-10、TNF-α应答特征

不同时间段采集猪的血液样本,利用ELISA方法检测血清和细胞培养上清中IL-12、IL-10、TNF-α等3种细胞因子的水平,分析猪双重免疫CSFV、PRRSV弱毒苗后血清和细胞培养上清中细胞因子的变化规律。结果显示,IL-12在血清中的浓度显著高于细胞培养上清中的浓度（P〈O．01）,在CSFV高抗体PRRSV高抗体组中28d采血的细胞培养上清中IL-12的浓度高于14d采血的细胞培养上清中的浓度（P〈0．05）}IL-10血清中水平显著低于细胞培养上清中水平（P〈0．01）,在CSFV高抗体PRRSV高抗体组中28d采血的细胞培养上清中的IL-10浓度显著低于14d采血的细胞培养上清中的浓度（P〈0．01）;TNF-α在血清中水平显著低于细胞培养上清中水平（P〈0．01）,在3组中28d采血的细胞培养上清液中TNF-α浓度高于14d采血的细胞培养上清液中浓度（P〈0．05）。结果表明,2种疫苗特异性抗体应答高时,IL-12水平显著上调,IL-10水平显著下调,提示上调IL-12及下调IL-10都是基因缺失疫苗发挥效力的潜在机制。TNF-α总体水平上升,但在血清中水平较低。     

The effect of dose and line on immunisation of cattle with lymphoblastoid cells infected with Theileria annulata

Thirty-one Friesian calves in Morocco susceptible to tropical theileriosis were protected against a lethal sporozoite challenge by prior infection with lymphoblastoid cell lines infected and transformed in vitro by a Moroccan stock of Theileria annulata. The challenge infection of cryopreserved sporozoites killed all four susceptible control calves within 20 days. Four schizont-infected cell cultures at Passage 3 were inoculated at four different doses, 10(8), 10(6), 10(4) and 10(2), into pairs of calves. The recipient animals showed great variation in severity of disease symptoms, which did not show a linear correlation with the cell dose inoculated. The most severe disease symptoms were recorded, prior to challenge, in the 10(2) cell dose recipients; one animal died of acute theileriosis and another had to be treated. One of the four cell lines used was more virulent than the other three. Two years after the completion of this experiment, immunised animals have shown normal productivity traits.     

NMDA对培养的大鼠腺垂体细胞分泌生长激素影响的研究

应用细胞培养、结合放射免疫测定法探讨了 N-甲基 -D-天冬氨酸 (NMDA)对培养的大鼠腺垂体细胞分泌生长激素 (GH)的影响。结果表明 :在腺垂体细胞培养中 ,试验组 (分别添加1 0 - 8、1 0 - 6 、1 0 - 4M NMDA)培养液中的 GH比对照组分别提高了 67.92 % ,87.46% ,1 0 0 .67% ,差异显著 (P <0 .0 5)。试验表明 NM-DA能直接刺激离体腺垂体细胞分泌 GH,并与NMDA呈剂量依赖关系     

稳定表达IL-10的PK-15细胞的构建及其在PCV2增殖中的应用

【目的】 研究白细胞介素-10（IL-10）对猪圆环病毒2型（Porcine circovirus type 2,PCV2）复制的影响,筛选PCV2高感染性细胞系和提高PCV2病毒滴度,为后续疫苗的研发及IL-10在PCV2感染中的作用研究提供参考。【方法】 利用PCR技术扩增猪IL-10基因,将目的基因与慢病毒表达载体（pCDH-CMV-MCS-EF1-GFP+Puro）进行连接,获得重组质粒pCDH-CMV-IL-10,将其与包装质粒psPAX2和pMD2.G共转染293T细胞进行慢病毒包装。用收集的慢病毒液感染PK-15细胞,经嘌呤霉素筛选后得到细胞株PK-15-IL-10,对照组细胞分别命名为PK-15-pCDH和PK-15。PCV2感染PK-15-IL-10、PK-15-pCDH和PK-15细胞株后,在24、48和72 h分别收集细胞液,利用CCK-8检测细胞活力。利用实时荧光定量PCR和Western blotting检测IL-10基因的表达水平和PCV2的复制情况;利用间接免疫荧光试验（IFA）观察PCV2在细胞中的复制情况及测定PCV2的病毒滴度（TCID₅₀）。【结果】 试验成功构建了重组质粒pCDH-CMV-IL-10,将其与包装质粒psPAX2和pMD2.G共转染293T细胞后,48 h时细胞状态最好,荧光最强。分别收集共转染48和72 h的慢病毒液上清感染PK-15细胞,pCDH-CMV-IL-10组的荧光最强,将其在嘌呤霉素浓度为2.5 μg/mL的完全培养基中继续培养,获得仍有绿色荧光的稳转细胞株。实时荧光定量PCR和Western blotting检测发现,IL-10基因在pCDH-IL-10细胞株中的表达量明显高于对照组PK-15-pCDH和PK-15,PCV2的拷贝数增加了4倍,复制能力增强,且将病毒稀释连续传3代后,PK-15-IL-10细胞中的PCV2极显著高于PK-15细胞（P<0.01）。细胞增殖试验表明,猪IL-10基因在细胞中过表达对细胞活力无明显影响;IFA结果表明,PK-15-IL-10细胞中的荧光比PK-15细胞更强,PCV2在PK-15-IL-10细胞中的TCID₅₀在感染后48 h极显著高于PK-15细胞（P<0.01）。【结论】 本研究成功构建了pCDH-CMV-IL-10的慢病毒表达载体,并利用其感染PK-15细胞,继续培养后筛选出过表达IL-10的PK-15-IL-10细胞株,用PCV2感染该细胞株能促进PCV2在PK-15细胞中的复制。本试验结果为后期疫苗研究提供了参考,为进一步研究IL-10对PCV2在PK-15细胞中复制的影响奠定了基础。     

Beclin-1基因shRNA慢病毒载体的构建及其对B16F10细胞自噬及活力的影响

利用RNA干扰技术构建稳定沉默Beclin-l基因的B16F10小鼠黑色素瘤细胞系，分析稳转细胞系的自噬水平和活力，为探讨Beclin-1基因、自噬与肿瘤三者关系奠定基础。构建小鼠Beclin-l基因干扰载体，转染至293T细胞，获得慢病毒颗粒，将病毒颗粒感染B16F10细胞，通过加压筛选、有限稀释、细胞驯化得到稳定转染m-Beclin-1-shRNA1的B16F10细胞系。采用RT-PCR和免疫荧光技术检测对Beclin-l的干扰效果，Western blot和透射电镜分析稳转细胞系中自噬标志物LC3蛋白表达及自噬小体形成情况，CCK-8法检测稳转细胞系的活力。结果表明，成功构建了靶向抑制Beclin-l基因表达的shRNA，纯化的慢病毒滴度为1×10⁸ TU·mL^-1，建立的细胞系中Beclin-1 mRNA和蛋白表达都受到不同程度的抑制，mRNA的沉默效率约为75%（P<0.01），发现干预Beclin-1表达能够抑制LC3-Ⅱ蛋白的表达及自噬小体的形成数量，降低B16F10细胞活力。以上结果表明，干扰Beclin-1表达能够有效降低B16F10细胞自噬活性，促进B16F10细胞死亡，这为探讨Beclin-1基因功能及自噬在抗肿瘤中的作用奠定重要基础。