动物细胞因子研究进展

细胞因子是免疫活性细胞和其它细胞分泌的具有多种生物活性的多肽或蛋白质 ,这些细胞因子是重要的信息传递物质 ,特别在免疫、炎症和造血系统等生物学反应过程中具有重要的作用。近几年 ,细胞因子的研究已成为生物学研究中最活跃的领域之一。新的细胞因子不断被发现 ,大量的细胞因子正在或将要应用于医学临床。细胞因子的研究带动了整个免疫学的发展。其应用展现出广阔的前景。文章简要介绍了动物细胞因子的作用及其应用前景 ,包括细胞因子与免疫抑制的关系、细胞因子在免疫增强剂和免疫治疗剂、构建新型基因工程苗以及细胞因子对于蛋白质、脂肪和葡萄糖代谢的调控作用等方面的主要研究进展     

Analysis of blood biochemistry and arterial blood gas of systemic inflammatory response syndrome in the early time after trauma

AIM: To explore the variation of blood biochemistry and arterial blood gas of patients with systemic inflammatory response syndrome (SIRS) in the early time after trauma and improve the diagnosis and first aid. METHODS: Eighty-eight patients with trauma from August 2003 to February 2004 were divided into two groups by their AIS-ISS90 score. The data of temperature, pulse, respiratory rate, white blood cell counts, Hb, blood glucose and arterial blood gas (PaO₂, PaCO₂, HCO₃^-, AG) were collected and compared with each group by statistic methods. RESULTS: Of the 88 patients, 49 underwent SIRS, 12 in light trauma group (ISS≥16) and 37 in severe trauma group (ISS<16). Compared with light trauma group, the data of pulse, respiratory rate, white blood cell counts, blood glucose, AG and rate of SIRS of severe trauma group were higher, PaO₂ and HCO₃^- were lower and the cases of PaCO₂>45 mmHg or <35 mmHg were more (P<0.01). The data of temperature and Hb had not significant difference between two groups (P>0.05). 13 patients had MODS in severe trauma group and 2 died while none had MODS or died in light trauma group. CONCLUSION: Application of AIS-ISS90 and SIRS-related blood biochemistry and arterial blood gas is beneficial for the diagnosis and treatment of patients in the early time after trauma.     

细胞因子对小尾寒羊胎盘成熟的影响

为了研究细胞因子对小尾寒羊胎盘成熟的影响 ,本实验采用液相竞争法和平衡法 ,对小尾寒羊空怀期 (n=5 )和妊娠期 (n=13)第 85天、10 5天、12 5天、14 0天和 15 0天 (足月 )时的血清白细胞介素 - 1β(IL - 1β)、白细胞介素 - 6 (IL - 6 )和肿瘤坏死因子(TNF)的含量分别进行检测。结果表明 ,IL - 1β和 IL - 6含量在 12 5天时与对照组间差异极显著 (P<0 .0 1) ,TNF含量在各时期与对照组差异均显著 (P<0 .0 1或 P<0 .0 5 )。随着妊娠过程的进展 ,三种细胞因子含量逐渐增加 ,12 5天时达到最高值 ,分别为 0 .390± 0 .196 ng/ ml,5 79.8± 15 2 .8pg/ ml和 2 .348± 0 .396 ng/ ml。而后逐渐下降 ,到足月前略有回升 ,但仍低于 12 5天时的最高值。由此看出小尾寒羊在妊娠期间 ,细胞因子与妊娠之间有着密切的关系 ,前者对于维持妊娠起到重要作用。由于 14 0天至足月3种细胞因子基本稳定 ,这表明胎盘于 14 0天左右达到成熟 ,IL- 1β和 IL- 6与启动分娩有关 ,而 TNF可能参与小尾寒羊分娩的启动。     

Pharmacokinetics of multiple doses of transdermal flunixin meglumine in adult Holstein dairy cows

A transdermal formulation of the nonsteroidal anti‐inflammatory drug, flunixin meglumine, has been approved in the United States and Canada for single‐dose administration. Transdermal flunixin meglumine was administered to 10 adult Holstein cows in their second or third lactation at the label dose of 3.33 mg/kg every 24 hr for three total treatments. Plasma flunixin concentrations were determined using high‐pressure liquid chromatography with mass spectroscopy (HPLC ‐MS ). Pharmacokinetic analysis was completed on each individual animal with noncompartmental methods using computer software. The time to maximum drug concentration (T max) was 2.81 hr, and the maximum drug concentration was 1.08 μg/ml. The mean terminal half‐life (T½) was determined to be 5.20 hr. Clearance per fraction absorbed (Cl/F) was calculated to be 0.294 L/hr kg^?1, and volume of distribution of fraction (V z/F ) absorbed was 2.20 L/kg. The mean accumulation factor was 1.10 after three doses. This indicates changes in dosing may not be required when giving multiple doses of flunixin transdermal. Further work is required to investigate the clinical efficacy of transdermal flunixin after multiple daily doses.     

Effects of continuous low dose infusion of lipopolysaccharide on inflammatory responses,milk production and milk quality in dairy cows

The objective of this study was to evaluate the effects of continuous low dose infusion of lipopolysaccharide (LPS) on inflammatory responses and milk production and quality in lactating dairy cows. Eight Holstein cows were assigned to two treatments in a cross‐over experimental design. Cows were infused intravenously either with saline solution or with saline solution containing LPS from Escherichia coli O111:B4 at a dose of 0.01 μg LPS/kg body weight for approximately 6 hr each day during a seven‐day trial. The clinical symptoms and milk production performance were observed. Milk samples were analysed for conventional components, fatty acids and amino acids. And jugular vein and mammary vein plasma samples were analysed for concentrations of cytokines and acute phase proteins. LPS infusion decreased feed intake and milk yield. An increase in body temperature was observed after LPS infusion. LPS infusion also increased plasma concentrations of interleukin‐1β, serum amyloid A, LPS‐binding protein, C‐reactive protein and haptoglobin. LPS infusion decreased the contents of some fatty acids, such as C17:1, C18:0, C18:1n9 (trans) and C18:2n6 (trans), and most amino acids except for methionine, threonine, histidine, cysteine, tyrosine and proline in the milk. The results indicated that a continued low dose infusion of LPS can induce an inflammatory response, decrease milk production and reduce milk quality.     

Lipopolysaccharide and cytokines modulate leukotriene (LT)B4 and LTC4 production by porcine endometrial endothelial cells

Uterine inflammatory response is mediated by inflammatory mediators including eicosanoids and cytokines produced by immune and endometrial cells. Interactions between lipopolysaccharide (LPS) and cytokines, and leukotrienes (LTs) in endothelium, important for the host defence during the inflammation, are unknown. We studied the effect of LPS, tumour necrosis factor (TNF)‐α, interleukin (IL)‐1β, IL‐4 and IL‐10 on 5‐lipooxygenase (5‐LO), LTA₄ hydrolase (LTAH) and LTC₄ synthase (LTCS) mRNA and protein expression, LTB₄ and LTC₄ release from porcine endometrial endothelial cells, and cell viability. For 24 hr, cells were exposed to LPS (10 or 100 ng/ml of medium) and cytokines (each 1 or 10 ng/ml). 5‐LO mRNA/protein expression augmented after incubation with larger doses of LPS, TNF‐α, IL‐4 and IL‐10 and smaller dose of IL‐1β. Larger dose of TNF‐α, smaller doses of LPS and IL‐1β and both doses of IL‐10 increased LTAH mRNA/protein expression. LTAH protein content was up‐regulated by larger dose of LPS, but it was reduced in response to both doses of IL‐4. LTCS mRNA expression was elevated by larger doses of LPS, IL‐4 and IL‐10 or both doses of TNF‐α and IL‐1β. LTCS protein level increased after treatment with both doses of IL‐1β, IL‐4 and IL‐10, smaller dose of LPS and larger dose of TNF‐α. Both doses of LPS and larger doses of TNF‐α and IL‐10 increased LTB₄ release. LPS, IL‐1β and IL‐10 at smaller doses, or TNF‐α and IL‐4 at larger doses stimulated LTC₄ release. Smaller doses of TNF‐α and IL‐1β or both doses of IL‐4 enhanced the cell viability. This work provides new insight on the participation of LPS, TNF‐α, IL‐1β, IL‐4 and IL‐10 in LTB₄ and LTC₄ production/release from porcine endometrial endothelial cells, and the effect of above factors on these cells viability. The used cellular model gives the possibility to further establish the interactions between inflammatory mediators.     

巨型艾美耳球虫偏菱形样蛋白EmRP免疫原性分析

在前期研究中,从巨型艾美耳球虫(Eimeria maxima)cDNA文库中筛选到了免疫保护性抗原偏菱形样蛋白EmRP,本研究为揭示该抗原的免疫保护机理,进一步检测了其免疫原性。以E.maxima卵囊cDNA为模板,用RT-PCR技术扩增EmRP,并构建真核表达质粒pVAX1-EmRP,将其经腿部肌肉注射2周龄的雏鸡,3周龄加强免疫。分别于首次免疫后和加强免疫后1周,采集血清,用ELISA方法检测血清特异性IgG水平,用荧光定量PCR方法(qPCR)检测细胞因子水平,用流式细胞术检测CD4^+T淋巴细胞和CD8^+T淋巴细胞的比例,分析EmRP诱导的免疫反应。序列分析表明,EmRP含有偏菱形样蛋白超家族成员保守区域,为非跨膜蛋白,分子量约为28.4 kD。真核表达质粒pVAX1-EmRP免疫鸡后,与对照组相比,鸡体IFN-γ、IL-2、IL-10和IL-17等细胞因子转录水平显著提升;CD8^+和CD4^+ T细胞比例显著提高,而血清特异性IgG水平与对照组差异不显著。结果表明,EmRP诱导的免疫保护作用主要是由其诱导的细胞免疫反应实现的,可以作为研制E.maxima新型疫苗的候选抗原。     

金黄色葡菌球菌脂蛋白对M1型小鼠骨髓源巨噬细胞免疫作用的影响

本研究旨在阐明金黄色葡萄球菌脂蛋白对M1型小鼠骨髓源巨噬细胞免疫作用的影响，为金黄色葡萄球菌致病性研究提供理论参考。以金黄色葡萄球菌野生株SA113（WT SA113）和SA113 lgt：：ermB脂蛋白表达缺失菌株（SA113Δlgt株）体外感染M1型小鼠骨髓源巨噬细胞，分为3组：空白对照组、WT SA113感染组（MOI：3：1）、SA113Δlgt感染组（MOI：3：1）。采用ELISA法检测M1型小鼠骨髓源巨噬细胞中肿瘤坏死因子-α（TNF-α）、白细胞介素1β（IL-1β）、趋化因子（RANTES）和白细胞介素10（IL-10）的水平，实时荧光定量PCR检测Toll样受体2（TLR2）、Toll样受体4（TLR4）和含NLR家族Pyrin域蛋白3（NLRP3）基因的表达，免疫荧光法检测脂蛋白对M1型小鼠骨髓源巨噬细胞吞噬金黄色葡萄球菌作用的影响。结果显示，与空白对照组相比，WT SA113感染组和SA113Δlgt感染组均可显著上调M1型小鼠骨髓源巨噬细胞中TNF-α、RANTES、IL-10分泌量以及WT SA113感染组TLR2、NLRP3基因表达水平（P<0.05），而TLR4基因表达量显著降低（P<0.05）；与WT SA113感染组相比，SA113Δlgt感染组M1型小鼠骨髓源巨噬细胞中TNF-α、IL-1β、RANTES、IL-10分泌量以及TLR2（12 h除外）、NLRP3基因表达水平显著降低（P<0.05）。免疫荧光结果显示，M1型巨噬细胞对SA113Δlgt株的吞噬作用显著低于对WT SA113株的吞噬作用（P<0.05）。综上，金黄色葡萄球菌的脂蛋白在M1型小鼠骨髓源巨噬细胞中主要通过激活TLR2和NLRP3受体，诱导细胞因子TNF-α、IL-1β、RANTES和IL-10的产生和释放。     

线粒体DNA在先天性免疫中的作用

线粒体是细胞内重要的细胞器,具有多种功能,是细胞的能量工厂。线粒体含有自己的遗传物质线粒体DNA,线粒体DNA因其在氧化磷酸化中的作用而广为人知。近年来,越来越多的研究表明线粒体DNA可作为先天性免疫系统的激动剂,并在病原体感染和炎性疾病的病理发展中起重要作用。线粒体DNA在进入细胞质或细胞外环境后,可以激活多种先天性免疫系统的模式识别受体,从而触发促炎细胞因子分泌和Ⅰ型干扰素反应。因此,本文就线粒体DNA激活先天性免疫的机制以及其在病原体感染和相关疾病发生发展中的作用进行讨论、总结,以期为进一步深入开展线粒体DNA在病原体感染及相关疾病中的作用及其机制研究提供理论依据。     

Potential protective effects of thiamine supplementation on the ruminal epithelium damage during subacute ruminal acidosis

In ruminants, the ruminal epithelium not only has the function of absorbing nutrients but also is an important tissue to prevent harmful substances in the rumen from entering the blood circulation. Thus, the normal function of ruminal epithelium is critical for ruminants. However, subacute ruminal acidosis induced by high-concentrate diets often damages the barrier function of ruminal epithelium in ruminants. Recently, many studies have shown that dietary supplementation with thiamine is an effective method to alleviate subacute ruminal acidosis. In order to provide theoretical reference for the in-depth study of subacute ruminal acidosis and the application of thiamine in the future, this review introduces the effects of subacute ruminal acidosis on morphological structure, inflammatory response, and tight junction of ruminal epithelium. In addition, this paper summarizes the role of thiamine in maintaining ruminal epithelial function of ruminants during subacute ruminal acidosis challenge.