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1.
在建立犊牛原代肝细胞体外培养模型的基础上，通过培养液中添加不同浓度的非酯化脂肪酸（Nonesterifiedfatty acids，NEFAs），运用实时荧光定量PCR和ELISA方法，测定NEFAs对肝细胞中炎性因子TNFα、IL-6和IL-1β的mRNA表达及其活性的影响。结果显示，NEFAs作用肝细胞9h后，与对照组相比，高浓度NEFAs（1．8mmol／L）组肝细胞中TNFα、IL-1β的mRNA表达水平显著增加（P〈0．05），IL-6的mRNA表达水平在2．4retool／L时极显著增加（P〈0．01），低浓度NEFAs（0．6、1．2mmol／L）组肝细胞中TNFα、IL-1β和IL-6的mRNA表达水平无显著差异（P〉0．05）；并且TNFα、IL-1G的活性在NEFAs浓度达到1．8、2．4mmol／L显著高于对照组（P〈0．01），且呈剂量依赖性作用，IL-6的活性在在NEFAs浓度达到1．8mmol／L显著高于对照组（P〈0．01），在2．4mmol／L也高于对照组，但差异不显著。结果表明，高浓度NEFAs在-定程度上能引起或加剧奶牛肝细胞炎性反应，可能是导致产后奶牛肝功能炎性损伤主要因素。 相似文献
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《中国兽医学报》2016,(3):508-512
旨在探讨非酯化脂肪酸(nonestesterified fatty acids,NEFAs)对体外原代培养犊牛肝细胞胰岛素信号通路的影响及SIRT1在其中的调控作用。选择培养48h的原代犊牛肝细胞,添加不同浓度的NEFAs(0、0.6、1.2、1.8mmol/L),继续培养12h,胰岛素处理组在收样前用100nmol/L胰岛素处理30min,每个浓度3个重复。另外选取已培养48h的细胞,用SIRT1抑制剂Nicotinamide(10mmol/L)处理12h,胰岛素处理组在收样前用100 mmol/L胰岛素处理30min。运用免疫印迹Western blot方法,检测NEFAs和Nicotinamide对肝细胞胰岛素信号通路关键蛋白IR和Akt的磷酸化水平及SIRT1蛋白表达水平的影响。结果显示,与对照组相比,1.2、1.8 mmol/L NEFAs组胰岛素刺激的IR磷酸化水平显著降低(P0.01),而Akt的磷酸化程度在NEFAs浓度为0.6 mmol/L时即显著降低(P0.01),同时,SIRT1的蛋白表达水平在NEFAs为1.2、1.8mmol/L显著降低(P0.01);添加SIRT1抑制剂显著降低SIRT1的蛋白表达水平(P0.01),同时胰岛素刺激的Akt磷酸化水平也显著降低(P0.01)。结果表明,高浓度的NEFAs可以损伤体外原代培养肝细胞的胰岛素信号通路,SIRT1在NEFAs损伤的胰岛素信号通路中发挥重要作用。 相似文献
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NEFA对体外培养奶牛中性粒细胞TLR2/4介导的炎症信号通路的影响 总被引：1，自引：0，他引：1
《中国兽医学报》2015,(4):626-630
分离培养奶牛中性粒细胞(PMN),添加不同浓度的非酯化脂肪酸(NFEA),作用浓度分别为0.125、0.25、0.5、1和2mmol/L。采用实时荧光定量PCR(qRT-PCR)检测炎性通路相关分子基因表达水平,采用ELISA检测炎性因子含量。结果显示,与对照组相比,Toll样受体2(TLR2)、Toll样受体4(TLR4)和核转录因子NF-κBp65的mRNA表达水平逐渐增加,且0.5、1和2mmol/L NEFA处理组显著高于对照组(P0.05,P0.01);炎性细胞因子白介素1β(IL-1β)、白介素6(IL-6)和肿瘤坏死因子α(TNF-α)含量也随着NEFA作用浓度的增加而增加,并且0.5、1和2mmol/L NEFA处理组显著高于对照组(P0.05,P0.01)。这些结果表明高浓度NEFA(≥0.5mmol/L)可以激活中性粒细胞TLR2/4介导的炎症信号通路,显著增加IL-1β、IL-6和TNF-α的合成和释放,造成中性粒细胞的炎性损伤,进而影响奶牛免疫机能。 相似文献
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《中国兽医学报》2016,(3):513-517
通过体外添加一定浓度的非酯化脂肪酸(nonestesterifiedfatty acid,NEFAs)(1.8 mmol/L)处理犊牛原代肝细胞不同时间,运用Western blot和实时荧光定量PCR(qRT-PCR)方法,测定NEFAs对内质网应激(ER stress)标志分子IRE1α磷酸化水平和GRP78、CHOP、XBP-1的mRNA表达水平的影响。结果显示,随着NEFAs作用时间的增加,与0.5h组相比,IRE1α磷酸化水平逐渐升高,NEFAs处理小于5h时,IRE1α磷酸化水平呈时间依赖性增加。并在5h达到最高水平,极显著高于0.5h组(P0.01),此后,磷酸化水平降低。与0.5h组相比,GRP78、CHOP、XBP-1的mRNA的表达水平整体逐渐升高,NEFAs处理9h时极显著高于0.5h时组(P0.01)并达到最高水平。结果表明,一定浓度的NEFAs可诱导奶牛肝细胞发生内质网应激。 相似文献
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《中国兽医学报》2016,(5):833-838
本试验旨在探讨非酯化脂肪酸(Nonestesterified fatty acid,NEFAs)对体外原代培养犊牛肝细胞线粒体功能以及罗格列酮对NEFAs诱导的线粒体功能紊乱的影响。体外添加NEFAs(1.8mmol/L)处理犊牛原代肝细胞不同时间,运用Western blot和实时荧光定量PCR(qRT-PCR)方法检测线粒体呼吸链复合体IV(COX IV)蛋白水平及mRNA水平的影响。另外选取细胞,罗格列酮(10nmol/L)预处理细胞24h后添加NEFAs(1.8mmol/L)处理9h。运用Western blot和qRT-PCR方法检测PGC-1α、Mfn-2的蛋白表达水平及COX IV mRNA表达水平。结果显示,随着NEFAs作用时间的增加,与0h组相比,NEFAs处理5h以上时COX IV蛋白表达水平极显著降低(P0.01),处理7~12h时COX IV mRNA表达水平呈显著降低(P0.05)。添加罗格列酮后,罗格列酮可极显著增加COX IV mRNA水平,PGC-1α、Mfn-2蛋白表达水平(P0.01)。结果表明,NEFAs可诱导犊牛原代肝细胞发生线粒体功能紊乱,罗格列酮可缓解由NEFAs诱导的犊牛原代肝细胞线粒体功能紊乱。 相似文献
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游离脂肪酸对原代培养奶牛肝细胞脂代谢的影响 总被引：1，自引：0，他引：1
《吉林畜牧兽医》2017,(6)
本实验旨在探讨游离脂肪酸(Nonestesterified fatty acids,NEFA)对体外培养奶牛原代肝细胞脂代谢的影响。选择培养48 h的奶牛原代肝细胞,添加不同浓度的NEFA(0、0.6、1.2、2.4 mmol/L)继续培养12 h,每个浓度5个重复。运用免疫印迹western blot方法,检测NEFA对奶牛原代肝细胞脂代谢信号通路关键蛋白过氧化物酶体增殖物激活受体α(Peroxisome proliferator activated receptor-α,PPARα)和固醇调节元件结合蛋白-1c(Sterol regulatory element binding protein-1c,SREBP-1c)蛋白表达水平的影响。利用甘油三酯检测试剂盒检测NEFA对奶牛原代肝细胞内甘油三酯含量的影响。结果显示,与对照组相比,0.6、1.2和2.4 mmol/L NEFA组PPARα蛋白表达水平显著降低(p0.01),而SREBP-1c的蛋白表达水平在NEFA浓度为1.2和2.4 mmol/L时显著降低(p0.01),同时,细胞内甘油三酯的含量在NEFA浓度为0.6 mmol/L时即显著升高(p0.01),且呈浓度依赖性。结果表明,高浓度的NEFA可以损伤体外培养奶牛原代肝细胞脂代谢信号通路,增加肝细胞内甘油三酯的沉积。 相似文献
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为研究线粒体融合蛋白2(mitofusin 2,MFN2)对高β-羟丁酸(β-hydroxybutyrate,BHBA)活化NF-κB炎性通路的影响,体外培养奶牛肝细胞,添加不同浓度(0.0,1.2,2.4,4.8 mmol/L)的BHBA,并转染过表达MFN2的腺病毒,运用Western blot和qRT-PCR技术检测NF-κB炎性通路关键分子的基因和蛋白表达。结果显示,随着BHBA浓度的增加,IKBα和NF-κB p65的磷酸化水平以及IL-1β、IL-6和TNFα的mRNA表达均显著升高,MFN2的基因和蛋白表达水平则显著降低;过表达MFN2后,显著抑制了高BHBA诱导的IKBα和NF-κB p65磷酸化水平及IL-1β、IL-6、TNFαmRNA表达水平的升高。结果表明,在奶牛肝细胞中过表达MFN2可以显著抑制高BHBA活化的NF-κB炎性通路。 相似文献
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为了研究体外非酯化脂肪酸(NEFA)对奶牛子宫内膜上皮细胞基质金属蛋白酶(MMPs)、Ⅳ型胶原蛋白(COL-Ⅳ)和炎性因子(IL-1β、IL-6、IL-8、TNF-α)分泌的调节作用,进而探讨NEFA对奶牛胎衣不下(RFM)发生的影响,使用不同浓度的NEFA刺激牛子宫内膜上皮细胞,采用qPCR和Western blot检测细胞中MMP-2、MMP-9、COL-Ⅳ、IL-1β、IL-6、IL-8、TNF-α的表达水平。结果表明,与对照组相比,低浓度组和高浓度组的MMP-2和COL-Ⅳ的mRNA表达极显著升高(P<0.01),MMP-9的mRNA极显著下调(P<0.01);IL-1β、IL-6、IL-8、TNF-α的mRNA表达在高浓度组均极显著上调(P<0.01),在低浓度组,IL-6的mRNA被极显著上调(P<0.01),IL-8和TNF-α的mRNA被显著上调(P<0.05)。Western blot的结果显示,与对照组相比,高浓度组的MMP-2和COL-Ⅳ的蛋白表达极显著上升(P<0.01),MMP-9的蛋白表达显著下降(P<0.05)。结... 相似文献
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《中国兽医学报》2020,(2):379-385
为了探究菊苣酸(chioric acid,CA)对脂多糖(lipopolysaccharide,LPS)诱导的牦牛外周血单个核细胞(peripheral blood mononuclear cell,PBMC)炎性相关因子分泌及转录水平的调节作用,用Ficoll法分离牦牛PBMC,体外将PBMC与LPS和CA共同培养,通过CCK-8法筛选LPS最佳刺激浓度,ELISA试剂盒检测CA对炎性相关因子含量的影响,实时荧光定量PCR法检测炎性相关因子mRNA表达情况。结果表明,CCK-8法筛选的LPS最佳致炎质量浓度为1.0 mg/L。与对照组相比,CA处理组显著提高IL-10的含量(P<0.05),LPS处理组显著提高IL-6和IL-1β的含量(P<0.05),极显著提高了IL-8、TNF-a和INF-γ的含量(P<0.01);与LPS处理组相比,CA+LPS处理组IL-6、IL-8、IL-1β、TNF-a和INF-γ的含量均显著降低(P<0.05),但IL-10的含量显著升高(P<0.05)。同时,与对照组相比,CA处理组显著降低IL-1βmRNA表达(P<0.05),显著升高IL-10 mRNA表达(P<0.05),LPS处理组极显著升高IL-6、IL-8、IL-1β、INF-γ和TNF-αmRNA表达(P<0.01);与LPS组相比,CA+LPS处理组IL-6、INF-γ和TNF-αmRNA表达显著降低(P<0.05),IL-8和IL-1βmRNA表达极显著降低(P<0.01),IL-10 mRNA表达显著升高(P<0.05)。上述结果说明,CA可通过调节牦牛PBMC炎性因子的分泌及表达,从而发挥抗炎作用。 相似文献
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高酮血症造成奶牛中性粒细胞先天免疫机能受到抑制,本研究探讨β-羟丁酸(BHBA)是否抑制脂多糖(LPS)诱导的奶牛中性粒细胞核因子-κB(NF-κB)信号通路的激活。分离健康奶牛中性粒细胞,采用LPS(100 ng/mL)和不同浓度(0.5、1.0、2.0和4.0 mmol/L)BHBA作用于中性粒细胞,收集细胞,应用实时荧光定量PCR(qRT-PCR)检测中性粒细胞中白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)和NF-κBp65 mRNA表达水平,Western blot检测NF-κBp65蛋白表达水平,比色法检测核因子-κB抑制物激酶β(IKKβ)激酶活性,酶联免疫吸附试验(ELISA)法检测促炎细胞因子TNF-α、IL-6和IL-1β的分泌量。结果表明：与对照组(不进行BHBA和LPS处理)相比较,LPS组(单独LPS处理)中IL-1β、IL-6、TNF-α和NF-κBp65 mRNA表达水平和NF-κBp65蛋白表达水平极显著增加(P <0.01),IKKβ激酶活性极显著增强(P<0.01),IL-1β和TNF-α的分泌... 相似文献
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魏静 《四川畜牧兽医学院学报》2009,(4):28-32
在现代法律秩序中,商会自治规范是制定法的基础和必要的补充,甚至在某些方面替代了制定法;商会自治规范主要包括商会组织规范、行为规范、惩罚规范以及争端解决规范等;其效力仅及于其内部成员;商会自治规范和制定法之间存在冲突,但也存在整合的基础。 相似文献
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本文概述了猪的毛色类型、猪的毛色遗传模式,着重综述了猪毛色基因分子基础的研究进展,指出存在问题并就未来发展方向做了思考。 相似文献
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REASONS FOR PERFORMING STUDY: Centesis of the bicipital bursa using an 8.9 cm long spinal needle has been reported but the alternative of employing a 3.8 cm long hypodermic needle requires validation. OBJECTIVE: To compare the efficacy of 2 different methods of centesis of the bicipital bursa and to evaluate the usefulness of ultrasonographic imaging to determine the location of solution administered when centesis of the bursa is attempted. METHODS: For Trial 1, 6 clinicians, who had no previous experience of centesis of the bicipital bursa, attempted to inject a solution composed of an aqueous radiopaque contrast medium and physiological saline solution (PSS) into the bicipital bursae of 2/12 horses using the previously described distal approach to inject one bursa and a proximal approach to inject the contralateral bursa. The bicipital tendon and bursa were examined ultrasonographically before and after injection; and both shoulders were examined radiographically to identify the location of the medium. In Trial 2, another 6 clinicians, also with no previous experience of centesis, repeated Trial 1, using 6 horses, but the radiopaque contrast medium was mixed with air instead of PSS. RESULTS: Accuracy of centesis using the proximal approach was 39% and that of the distal approach 28%. Ultrasonographic examination of the shoulder allowed the location of solution and air to be accurately predicted in all 12 shoulders examined. CONCLUSIONS: Clinicians who have had no previous experience performing centesis of the bicipital bursa are unlikely to be successful in centesis using either approach. Radiographic examination after injecting a radiopaque contrast medium may be necessary to assess the success of centesis especially if bursal fluid is not obtained during centesis. Injecting air along with the radiopaque contrast medium provides more accurate ultrasonographic confirmation of centesis and better radiographic definition than does injection without air. 相似文献
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Effects of size of ingestively masticated fragments of plant tissues on kinetics of digestion of NDF
Ingestively masticated fragments were collected and sized via sieving. Different sizes of esophageal masticate and ruminal digesta fragments, and ground fragments of larger masticated pieces were incubated in vitro, and undigested NDF remaining at intervals of up to 168 h of incubation was determined. The ruminal age-dependent time delay (tau) for onset of digestion of NDF was positively correlated (P < 0.004) with the mean sieve aperture estimated to retain 50% of the fragments between successive sieve apertures (MRA). Degradation rate of potentially degradable NDF (PDF) and level of indigestible NDF were not related (P > 0.10) to MRA of masticated and ground fragments. Estimates of tau were positively related to MRA, with slopes of bermudagrass < corn silage < ruminal fragments of corn silage. It was concluded that fragment size-, and consequently, ruminal age-dependent onset of PDF degradation of a mixture of different fragment sizes results in an age-dependent rate of degradation of the more rapidly degrading of two subentities of PDF. Models are proposed that assume a tau before onset of simultaneous degradation of PDF from two pools characterized as having gamma-modeled age-dependency and age-constant rates. The ruminal age-dependent pool seems to be associated with the faster-degrading pool, and its rate parameter increases with range in MRA in the population of fragments. Conceptually, the ruminal age-dependent rate parameter for PDF degradation seems to represent a composite of several effects: 1) effects of the size-dependent tau; 2) range in MRA of the population of ingestively masticated fragments; and 3) subentities of PDF that degrade via more rapid age-dependent rates compared with subentities of PDF that degrade via age-constant rates. The estimated fractional rates of ruminative comminution of ingestively masticated fragments (0.060 to 0.075/h) were of a magnitude similar to the mean fractional rates of PDF digestion (0.030 to 0.085/h), which implies that ruminative comminution may be first-limiting to fractional rate of PDF digestion. The in vivo roles of ingestive and ruminative mastication of fragments on PDF degradation must be considered in any kinetic system for estimating PDF digestion in the rumen. These results and others in the literature suggest that the rate of surface area exposure rather than intrinsic chemical attributes of PDF may be first-limiting to degradation rate of PDF in vivo. 相似文献
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乳酸杆菌作为益生菌广泛用于人和动物.本文综述了乳酸杆菌改善宿主健康的机制.乳酸杆菌可通过产生抗菌物质如乳酸、过氧化氢、细菌素,或者通过竞争营养或肠道黏附位点来抑制致病菌;通过诱导黏附素的分泌或阻止细胞凋亡而增强肠道的屏障功能,从而保护肠道.文章重点讨论了乳酸杆菌表面成分(表面蛋白、脂磷壁酸和肽聚糖)与肠道受体(C型凝集素受体、Toll样受体和Nod样受体),阐述了他们结合后启动免疫调节信号,调控肠道免疫功能以发挥改善健康作用的机制. 相似文献
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Billinghurst RC Brama PA van Weeren PR Knowlton MS McIlwraith CW 《American journal of veterinary research》2004,65(2):143-150
OBJECTIVE: To determine whether serum concentrations of biomarkers of skeletal metabolism can, in conjunction with radiographic evaluation, indicate severity of osteochondrosis in developing horses. ANIMALS: 43 Dutch Warmblood foals with varying severity of osteochondrosis. PROCEDURE: 24 foals were monitored for 5 months and 19 foals were monitored for 11 months. Monthly radiographs of femoropatellar-femorotibial and tibio-tarsal joints were graded for osteochondral abnormalities. Serial blood samples were assayed for 8 cartilage and bone biomarkers. At the end of the monitoring period, foals were examined for macroscopic osteochondrosis lesions. RESULTS: Temporal relationships were evident between certain serum biomarkers and osteochondrosis severity in foals during their first year. Biomarkers of collagen degradation (collagenase-generated neoepitopes of type-II collagen fragments, type-I and -II collagen fragments [COL2-3/4C(short)], and cross-linked telopeptide fragments of type-I collagen) and bone mineralization (osteocalcin) were positive indicators of osteochondrosis severity at 5 months of age. In foals with lesions at 11 months of age, osteochondrosis severity correlated negatively with COL2-3/4C(short) and osteocalcin and positively with C-propeptide of type-II procollagen (CPII), a collagen synthesis marker. Radiographic grading of osteochondrosis lesions significantly correlated with macroscopic osteochondrosis severity score at both ages and was strongest when combined with osteocalcin at 5 months and CPII at 11 months. CONCLUSIONS AND CLINICAL RELEVANCE: The ability of serum biomarkers to indicate osteochondrosis severity appears to depend on stage of disease and is strengthened with radiography. In older foals with more permanent lesions, osteochondrosis severity is significantly related to biomarker concentrations of decreased bone formation and increased cartilage synthesis. 相似文献