Porcine hepatocyte–Kupffer cell co‐culture as an in vitro model for testing the efficacy of anti‐inflammatory substances          下载免费PDF全文

G. Mátis  A. Kulcsár  J. Petrilla  P. Talapka  Z. Neogrády 《Journal of animal physiology and animal nutrition》2017,101(2):201-207

As Kupffer cells are highly involved in the regulation of hepatic inflammatory response, the main goal of this study was to improve and to characterize a hepatocyte–Kupffer cell co‐culture of pig origin for modelling endotoxin‐induced hepatic inflammation and for testing the efficacy of potential anti‐inflammatory substances. This monolayer co‐culture was prepared from primary isolated swine hepatocytes and Kupffer cells in the ratio of 6:1 and 2:1, mimicking different states of liver inflammation. The prepared cell cultures were characterized by immunohistochemical CD‐68 detection. Lipopolysaccharide (LPS) challenge of both co‐cultures resulted in elevated interleukin‐8 (IL‐8) and that of 6:1 co‐cultures in increased IL‐6 production with a higher extent than on hepatocyte monocultures, justifying the key role of Kupffer cells in pro‐inflammatory cytokine production. LPS‐induced IL‐8 production was successfully attenuated by concomitant application of both sodium butyrate and terpinen‐4‐ol on hepatocyte monocultures, but not on co‐cultures, demonstrating the importance of the presence of Kupffer cells in cell cultures as inflammatory models. Based on these initial data, the applied porcine primary hepatocyte–Kupffer cell co‐culture is suggested to be a proper tool for in vitro investigations on liver physiology and hepatic inflammation in pigs and can be used as a useful model mimicking in vivo conditions in veterinary research.  相似文献   

检测细胞因子对监测鸡免疫抑制病的临床价值     

马振乾  王金良  郭振环  于新友  何剑斌  沈志强 《中国畜牧兽医》2011,38(10):185-187

细胞因子是指一些由机体免疫或非免疫细胞产生的具有生物活性,与造血、炎症和免疫应答反应密切相关的小分子多肽类糖蛋白。因此,在动物免疫抑制病中对细胞因子的检测有着非常重要的临床价值。文章介绍了细胞因子的检测方法、与免疫抑制相关的细胞因子及细胞因子检测在研究动物免疫抑制病中的价值。  相似文献   

细胞因子在奶牛乳房炎生物学防治中的应用     

王玲  李宏胜  陈炅然  苗小楼  王正兵  尚若锋 《中国畜牧兽医》2011,38(12):173-176

鉴于细胞因子对乳房免疫机能独特有效的操纵和调节,在奶牛乳房炎控制中具有一定独特优势,作者就细胞因子的生物学活性、作用机制和特点进行了综述,并重点对国内外应用奶牛重组细胞因子防治奶牛乳房炎进行了总结、分析,有望为控制奶牛乳房炎提供安全、有效、实用的生物学防治途径。  相似文献   

Interferon-gamma, interleukin-4 and interleukin-10 production by T helper cells reveals intact Th1 and regulatory TR1 cell activation and a delay of the Th2 cell response in equine neonates and foals     

Bettina Wagner  Alexandra Burton    Dorothy Ainsworth 《Veterinary research》2010,41(4)

Cytokines produced by T helper (Th) cells are important in orchestrating the immune response during health and disease. Recent reports indicated that cytokine mRNA expression in foals is often quantitatively lower than that of adult horses suggesting that foal T cells are not fully mature. Here, peripheral blood mononuclear cells from foals and adult horses were stimulated with phorbol 12-myristate 13-acetate and analyzed for intracellular interferon-gamma (IFN-γ), interleukin-4 (IL-4) and IL-10 production, representing the Th1, Th2 and regulatory T_R1 cell phenotypes respectively, by flow cytometry. In agreement with previous reports, all three cytokines were quantitatively reduced in foals compared to adults. However, the balance between Th1 and Th2 cytokines (IFN-γ/IL-4 ratio) showed a clear Th1-biased response in foals by 6 and 12 weeks of life, while similar IFN-γ/IL-10 ratios were found in foals and adult horses. By day 5 after birth, intracellular IFN-γ production by foal CD4⁺ and CD8⁺ T cells resembled that in adults. Overall, IL-4 production was low in foals. IL-4⁺ cells peaked at day 5 of age when IL-4 was mainly produced by IgE⁺ cells. Relative percentages of IL-4⁺ Th2 cells were significantly lower in foals at all time points. The data suggested that equine neonates and young foals have an impaired Th2 response, that the immune response of foals is Th1 biased, that IFN-γ production by Th and cytotoxic T cells is qualitatively similar to adult horses, and regulatory IL-10 production by T cells is developmentally mature in foals during the first three months of life.  相似文献   

Induction of apoptotic lesions in liver and lymphoid tissues and modulation of cytokine mRNA expression by acute exposure to deoxynivalenol in piglets     

Osamu Mikami  Hiroyuki Yamaguchi  Hideo Murata  Yasuyuki Nakajima  Shigeru Miyazaki 《Journal of veterinary science (Suw?n-si, Korea)》2010,11(2):107-113

Six 1-month-old piglets were intravenously injected with deoxynivalenol (DON) at the concentration of 1 mg/kg body weight, with three pigs each necropsied at 6 and 24 h post-injection (PI) for investigation of hepatotoxicity and immunotoxicity with special attention to apoptotic changes and cytokine mRNA expression. Histopathological examination of the DON-injected pigs revealed systemic apoptosis of lymphocytes in lymphoid tissues and hepatocytes. Apoptosis of lymphocytes and hepatocytes was confirmed by the TdT-mediated dUTP-biotin nick end-labeling (TUNEL) method and immunohistochemical staining against single-stranded DNA and cleaved caspase-3. The number of TUNEL-positive cells in the thymus and Peyer''s patches of the ileum was increased at 24 h PI compared to 6 h PI, but the peak was at 6 h PI in the liver. The mRNA expression of interleukin (IL)-1β, IL-6, IL-18, and tumor necrosis factor (TNF)-α in the spleen, thymus and mesenteric lymph nodes were determined by semi-quantitative RT-PCR, and elevated expression of IL-1β mRNA at 6 h PI and a decrease of IL-18 mRNA at 24 h PI were observed in the spleen. IL-1β and IL-6 mRNA expressions increased significantly at 6 h PI in the thymus, but TNF-α decreased at 6 h PI in the mesenteric lymph nodes. These results show the apoptosis of hepatocytes suggesting the hepatotoxic potential of DON, in addition to an immunotoxic effect on the modulation of proinflammatory cytokine genes in lymphoid organs with extensive apoptosis of lymphocytes induced by acute exposure to DON in pigs.  相似文献   

Effects of rosiglitazone on expression of SOCS1/SOCS3 and inflammatory reaction in RAW 264.7 cell-derived foam cells     

LI Fei  YUAN Yong  DONG Yu-gang  DONG Jian-ting  FENG Li  DENG Zhi-hua 《园艺学报》2013,29(2):261-265

AIM: To investigate whether perioxisome proliferator-activated receptor γ (PPARγ) ligand rosiglitazone regulates suppressor of cytokine signaling 1 (SOCS1) and SOCS3 expression as well as pro-inflammatory/anti-inflammatory responses in RAW 264.7 cell-derived foam cells. METHODS: The concentrations of TNF-α, IL-6 and IL-10 in the cultured supernatant of RAW 264.7 cell-derived foam cells were detected by ELISA, and the ratios of TNF-α/IL-10 and IL-6/IL-10 were calculated. RT-PCR and Western blotting were used to analyze the effects of rosiglitazone on the expression of SOCS1 and SOCS3 at mRNA and protein levels. RESULTS: The concentrations of TNF-α, IL-6 and IL-10, and ratios of TNF-α/IL-10 and IL-6/IL-10 in foam cell group were obviously higher than those in control group, but the concentrations of the above factors in oxidized low-density lipoprotein (ox-LDL) +rosiglitazone group were apparently lower than those in foam cell group. The expression of SOCS1 and SOCS3 at mRNA and protein levels in oxLDL+rosiglitazone group was apparently higher than that in control and foam cell group. CONCLUSION: PPARγ ligand rosiglitazone up-regulates the expression of SOCS1 and SOCS3 at mRNA and protein levels and regulates the balance of pro-inflammatory/anti-inflammatory responses in RAW 264.7 cell-derived foam cells.  相似文献   

副猪嗜血杆菌外膜蛋白P5对豚鼠树突状细胞成熟及功能影响     

佘志成  范红结  李春玲  王贵平  贾爱卿  杨冬霞 《畜牧兽医学报》2011,42(7)

为研究副猪嗜血杆菌(Haemophilus parasuis,Hps)外膜蛋白P5对豚鼠骨髓来源的树突状细胞(Dendritic cell,DC)刺激淋巴细胞增殖功能及细胞因子分泌的影响,以GM-CSF、IL-4联合诱导骨髓细胞生成未成熟DC,加入不同剂量副猪嗜血杆菌外膜蛋白P5,观察DC形态,检测混合淋巴细胞反应,ELISA法测IL-6、IL-12p70、TNF-α分泌情况.结果显示:经副猪嗜血杆菌外膜蛋白P5刺激后,可获得具有典型树突状突起形态的DC;经5μg·mL-1P5刺激的DC较对照IL-6、TNF-α分泌量极显著增加(P＜0.01),IL-12的分泌量有所增加(P＞0.05);50 μg·mL-1P5刺激的DC较对照IL-6分泌量极显著增加(P＜0.01),IL-12p70和TNF-α分泌量均极显著下降(P＜0.01).诱导后的DC刺激同种异体T淋巴细胞增殖的能力极显著增强(P＜0.01).本研究证明P5能影响骨髓细胞来源的DC的分化、成熟及功能的发挥,且不同剂量的P5对DC的成熟程度影响有差异.  相似文献   

紫花地丁总黄酮体外抗炎活性研究     

张静  邵永斌  谷新利  罗燕 《中国畜牧兽医》2020,47(4):1258-1266

试验研究了紫花地丁总黄酮（TFV）对脂多糖（LPS）诱导的小鼠RAW264.7巨噬细胞活力、细胞中炎症介质含量以及相关基因表达的影响,以期探讨其体外抗炎活性的作用。试验采用MTT法筛选出TFV对小鼠RAW264.7巨噬细胞活力具有促进作用的最佳添加浓度;用酶联免疫吸附法（ELISA）检测了TFV对LPS诱导的小鼠RAW264.7巨噬细胞释放到细胞培养液中NO、肿瘤坏死因子α（TNF-α）、白介素1β（IL-1β）、白介素6（IL-6）含量的影响;运用实时荧光定量PCR法检测了TFV对LPS诱导的炎性小鼠RAW264.7巨噬细胞TNF-α、诱导型一氧化氮合酶（iNOS）和环氧合酶2（COX-2）相对表达水平的影响;研究并分析了TFV的体外抗炎活性。试验结果表明,TFV在5~50 μg/mL浓度范围内能提高小鼠RAW264.7巨噬细胞的活力（P<0.05）;与LPS模型组比较,TFV能显著降低LPS诱导的小鼠RAW264.7巨噬细胞产生NO、TNF-α、IL-6、IL-1β的含量,并能显著降低LPS诱导的小鼠RAW264.7巨噬细胞内TNF-α、COX-2等炎症因子的mRNA表达量（P<0.05）。综上,TFV能显著下调LPS诱导的小鼠RAW264.7巨噬细胞IL-1β、IL-6、TNF-α等细胞因子的释放量和下调TNF-α、COX-2 mRNA的表达量,说明抑制促炎性细胞因子基因的表达可能是实现其抗炎作用的原因之一。  相似文献   

奶牛乳腺免疫细胞防御机理研究进展     

赵悦  童津津  熊本海  蒋林树 《动物营养学报》2020,(4):1563-1569

奶牛乳腺中的初级吞噬细胞、嗜中性粒细胞(PMN)和巨噬细胞构成了抵御病原体入侵的第1道防线。在健康奶牛的乳腺中,巨噬细胞占主导地位且充当哨兵的角色。当病原体侵入乳腺时,巨噬细胞及乳腺上皮细胞就会释放出直接将PMN迁移到该区域的趋化剂,使PMN从循环中迅速流入并吞噬和杀灭细菌,起到保护的作用。抵御病原体入侵的第2道防线是由记忆细胞和免疫球蛋白组成的网络,它们与第1道防线相互作用。随着分子生物学技术的发展,更好地了解炎症反应的调节机制可为研究和调节宿主与病原体的相互作用提供理论基础,因此本文就奶牛乳腺免疫细胞防御机理的研究进展进行了综述。  相似文献   

Immunoexpression of cytokine tumour necrosis factor-α suggesting its role in formation and regression of corpus luteum in Indian buffalo     

Kritima Kapoor  Opinder Singh  Devendra Pathak 《Reproduction in domestic animals》2020,55(10):1393-1403

Tumour necrosis factor-α (TNF-α) is a cytokine that plays multiple important roles in corpus luteum (CL). Immunolocalization of expression of TNF-α in CL of buffalo was studied in different stages of its development and regression. Corpus luteum of healthy buffaloes (24) was collected from local slaughterhouses and categorized into early (stage I, 1–5 days, n = 6), mid (stage II, 6–11 days, n = 6), late luteal (stage III, 12–16 days, n = 6) and regressing phase (stage IV, 17–20 days, n = 6). In earliest phase of cyclic CL, per cent immunoexpression of TNF-α was significantly (p < .05) lower as compared to all phases with its expression being restricted to few developing luteal cells, usually in neutrophils. A significantly (p < .05) higher number of neutrophils with TNF-α immunoexpression were observed as compared to mid-luteal phase that indicated its role in initiation of angiogenesis at this stage. TNF-α immunoexpression almost doubled in mid-luteal phase, but the number of neutrophils exhibiting TNF-α was significantly (p < .05) lower with respect to all phases of CL. Immunoexpression percentage in late luteal phase increased sharply being significantly (p < .05) higher than earlier two phases of CL. In regressing phase, per cent immunostaining was maximum with highly significant (p < .05) difference as compared to all other stages, observed in all degrading luteal cells, abundant immune cells, that is neutrophils and macrophages which finally led to apoptosis and phagocytosis. Immunoexpression of TNF-α in early luteal phases served its role in initiation of angiogenesis, and its intense expression in regressing phase of CL suggested a shift in its role to apoptosis and structural luteal regression signifying both luteotropic and luteolytic roles in buffalo. This is probably the first study of its kind in buffaloes.  相似文献