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1.
β‐carotene is one of the most abundant carotenoids, has potential anti‐inflammatory effect, it has been reported that β‐carotene could suppress LPS‐induced inflammatory responses by inhibiting nuclear factor kappa B (NF‐κB) translocation, but the more detailed molecular mechanisms underlying the anti‐inflammatory action of β‐carotene remain to be fully understood. In this study, we investigated the influence of β‐carotene on the activation of JAK2/STAT3, MAPK, and NF‐κB signaling pathway induced by LPS in RAW264.7 cells and peritoneal macrophages. Cells were treated with different concentrations of β‐carotene for 3 hr after LPS treatment for 24 hr. The mRNA expression and the release of IL‐1β, IL‐6, and TNF‐α were evaluated by RT‐PCR and ELISA, and the level of signaling proteins of JAK2/STAT3, MAPK, and NF‐κB signaling pathway were detected by Western blot. The results showed that β‐carotene significantly suppressed (p < 0.05) LPS‐induced release of IL‐1β, IL‐6, and TNF‐α and their mRNA expression. LPS‐induced JAK2/STAT3, IκB/NF‐κB p65, JNK/p38 MAPK signal activation were significantly attenuated (p < 0.05) by β‐carotene in a dose‐dependent manner. In conclusion, β‐carotene could attenuate LPS‐induced inflammation via inhibition of the NF‐κB, JAK2/STAT3, and JNK/p38 MAPK signaling pathways in macrophages.  相似文献   

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NK‐lysins are cationic peptides that play important roles in host protection, and are an important constituent of innate immunity. We identified nine single‐nucleotide polymorphisms (SNPs) in the NK‐lysin open reading frame (ORF) from 32 Japanese quails in six strains: A, B, ND, K, P, and Y. The G to A substitution at nucleotide position 272 in the ORF resulted in a Gly (G) to Asp (D) amino acid substitution (Cj31G and Cj31D alleles). The Cj31D allele was detected in P (frequency 0.76) and Y (frequency 0.03) strains. We compared the antimicrobial activities of four synthetic peptides from the helix 2‐loop‐helix 3 region of avian NK‐lysins against Escherichia coli: Cj31G and Cj31D from quail and Gg29N and Gg29D from chicken. The antimicrobial activities of the four peptides decreased in the following order: Gg29N > Cj31G > Gg29D > Cj31D (P < 0.05). Although there were no differences in the predicted secondary structure of the Cj31G and Cj31D, the net charge of the Cj31G was higher than that of Cj31D. These data indicated that the antimicrobial activity of CjNKL is influenced by net charge, similar to that which has been observed in chicken. © 2015 Japanese Society of Animal Science  相似文献   

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PCR扩增鸡L-FABP基因5′侧翼区约2kb的DNA片段,进行克隆并测序,构建了鸡L-FABP基因报告基因系列缺失载体,瞬时转染进入人肝癌细胞系,利用双荧光素酶报告基因系统测定了荧光素酶活性。在线分析软件发现鸡L-FABP基因启动子区存在HNF-1、SREBP-1、AP-1、C/EBP、Oct-1、TATA、CCAAT、GATA-1等调控元件,没有发现CpG岛。报告基因结果表明鸡L-FABP基因启动子-2 076bp/-20bp区域具有最强的启动子活性,-522bp/-20bp区域启动子活性最弱;C/EBPα可以显著的抑制鸡L-FABP基因的表达,这些结果为深入研究鸡L-FABP的表达调控机制奠定了基础。  相似文献   

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Adiponectin is a protein hormone secreted exclusively by adipocytes that plays an important role in the modulation of glucose and lipid metabolism. To investigate the effect of adiponectin on lipid metabolism in chicken, rosiglitazone (agonist of adiponectin) and dexamethasone (inhibitor of adiponectin) were used to treat 23‐day‐old broilers in vivo. To verify the functionality of adiponectin on fat deposition, chicken pre‐adipocytes were cultured in the medium containing 10 μg/ml adiponectin. Serum adiponectin and lipids and fat distribution were analysed. Oil Red O staining was used to determine lipid deposition in adipocytes. The expression levels of adiponectin, adiponectin receptors (AdipoR) and lipid metabolism–related genes in different tissues and pre‐adipocytes were measured using real‐time PCR, and the abundance of lipid metabolism–related proteins was measured by Western blot. Rosiglitazone increased serum adiponectin concentration and the expression levels of adiponectin and adiponectin receptor 1 (AdipoR1) in tissues and significantly decreased levels of serum lipids and fat deposition. Rosiglitazone significantly increased the expression levels of adipose triglyceride lipase (ATGL) and AdipoR1 and decreased the expression levels of fatty acid synthase (FAS). Dexamethasone had the converse effects compared with rosiglitazone. Oil red O staining results showed a marked decrease in fat deposition in cells treated with adiponectin. In adipocytes, adiponectin could decrease the expression levels of CCAAT/enhancer‐binding protein α (C/EBPα) and FAS and increased the expression levels of ATGL and AdipoR1. These results indicate that adiponectin has a remarkable effect on impairment of adipocyte differentiation, which contributes to the negative regulation of fat deposition in chicken.  相似文献   

6.
Liver‐expressed antimicrobial peptide 2 (LEAP‐2) is a cationic peptide that plays an important role in innate immunity for host defense. The aim of this study was to characterize the LEAP‐2 gene in the Japanese quail (Coturnix japonica). Japanese quail LEAP‐2 (CjLEAP‐2) was identified from the Japanese quail draft genome database by a local BLAST analysis using chicken LEAP‐2 (GgLEAP‐2). The exon‐intron structure of CjLEAP‐2, analyzed from three quails, is composed of three exons, as is the chicken LEAP‐2 homolog (GgLEAP‐2). An analysis of the coding sequence revealed that CjLEAP‐2 is 231 bp long, like GgLEAP‐2, and 93% identical to GgLEAP‐2 at the nucleic acid level. The predicted amino acid sequence of CjLEAP‐2 contained the liver‐expressed antimicrobial peptide 2‐precursor domain and four cysteine residues characteristic of the LEAP‐2 protein. The amino acid sequence of the mature peptide of CjLEAP‐2 was 100% identical to that of GgLEAP‐2. We confirmed that CjLEAP‐2 was transcribed in at least seven tissues, including the digestive system. Additionally, the mature peptide region of CjLEAP‐2 exhibited no polymorphisms in 99 quails from six strains. Taken together, these findings indicate that CjLEAP‐2 is non‐polymorphic and therefore, it likely plays an important role in the innate immunity of quail as it does in chicken.  相似文献   

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Krüppel‐like factor 7 (KLF7) has been extensively studied in mammalian species, but its role in birds is still unclear. In the current study, cloning and sequencing showed that the full‐length coding region of chicken KLF7 (Gallus gallus KLF7, gKLF7) was 891 bp long, encoding 296 amino acids. In addition, real‐time RT‐PCR analysis showed that gKLF7 was broadly expressed in all 15 chicken tissues selected, and its expression was significantly different in spleen, proventriculus, abdominal fat, brain, leg muscle, gizzard and heart between fat and lean broilers at 7 weeks of age. Additionally, one novel single nucleotide polymorphism (SNP), XM_426569.3: c. A141G, was identified in the second exon of gKLF7. Association analysis showed that this locus was significantly associated with fatness traits in Arbor Acres broiler random population and the eighth generation of Northeast Agricultural University broiler lines divergently selected for abdominal fat content (NEAUHLF) population (P < 0.05). These results suggest that gKLF7 might be a candidate gene for chicken fatness traits.  相似文献   

13.
Au, A. Y., Hasenwinkel, J. M., Frondoza, C. G. Silybin inhibits interleukin‐1β‐induced production of pro‐inflammatory mediators in canine hepatocyte cultures. J. vet. Pharmacol. Therap. 34 , 120–129. Hepatocytes are highly susceptible to cytokine stimulation and are fundamental to liver function. We established primary canine hepatocyte cultures to study effects of anti‐inflammatory agents with hepatoprotective properties. Hepatocyte cultures were incubated with control media alone, silybin (SB), or the more bioavailable silybin–phosphatidylcholine complex (SPC), followed by activation with interleukin‐1 beta (IL‐1β; 10 ng/mL). Inflammatory response was measured by prostaglandin E2 (PGE2), interleukin‐8 (IL‐8), and monocyte chemotactic protein‐1 (MCP‐1) production and also nuclear factor‐kappa B (NF‐κB) translocation. Hepatocyte cultures continued production of the phenotypic marker albumin for more than 7 days in culture. IL‐1β exposure increased PGE2, IL‐8, and MCP‐1 production, which was paralleled by NF‐κB translocation from the cytoplasm to the nucleus. Pretreatment with SB and SPC significantly inhibited IL‐1β‐induced production of pro‐inflammatory markers and attenuated NF‐κB nuclear translocation. We demonstrate for the first time that primary canine hepatocyte cultures can be maintained in culture without phenotypic loss. The observation that hepatocyte cultures respond to pro‐inflammatory IL‐1β activation indicates hepatocytes as primary cellular targets of extrinsic IL‐1β. The ability of SB and SPC to inhibit hepatocyte culture activation by IL‐1β reinforces the notion of their hepatoprotective effects. Our primary canine hepatocyte culture model facilitates identification of hepatoprotective agents and their mechanism of action.  相似文献   

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The female sex hormone estrogen exerts anti‐inflammatory effects. The G‐protein‐coupled estrogen receptor (GPER) has been recently identified as a novel membrane‐type estrogen receptor that can mediate non‐genomic estrogenic effects on many cell types. We previously demonstrated that GPER inhibits tumor necrosis factor alpha‐induced expression of interleukin 6 (IL‐6) through repression of nuclear factor‐kappa B (NF‐κB) promoter activity using human breast cancer cells. Although several reports have indicated that GPER suppresses Toll‐like receptor‐induced inflammatory cytokine expression in macrophages, the molecular mechanisms of the inhibition of cytokine production via GPER remain poorly understood. In the present study, we examined GPER‐mediated inhibition of IL‐6 expression induced by lipopolysaccharide (LPS) stimulation in a mouse macrophage cell line. We found that the GPER agonist G‐1 inhibited LPS‐induced IL‐6 expression in macrophage cells, and this inhibition was due to the repression of NF‐κB promoter activity by GPER. G‐1 treatment also decreased the phosphorylation of inhibitor of κB kinases. Among the mitogen‐activated protein kinases, the phosphorylation of c‐jun N‐terminal kinase (JNK) was increased by G‐1. These findings delineate the novel mechanism of the inhibition of LPS‐induced IL‐6 through GPER‐activated JNK‐mediated negative regulation of the NF‐κB pathway in murine macrophage cells, which links anti‐inflammatory effects to estrogen.  相似文献   

15.
The duck growth hormone encoding gene and its promoter region were amplified by polymerase chain reaction (PCR). A total of 5.25 kb were cloned and sequenced. Duck growth hormone (GH) consists of five exons and four introns and is structurally similar to mammalian and chicken GH gene. Although the distal region of duck GH promoter showed no similarity to chicken and turkey promoters, the proximal region of the promoter contained two putative Pit‐1 binding sequences, and showed similarity to chicken and turkey GH promoters. Genetic variation was detected at five positions of the promoter region. The results of this study indicate that the expression of duck GH is likely regulated in a similar manner to that of chicken GH via enhancer‐type cis‐acting elements and the presence of genetic variation in the duck GH gene may be applicable to marker‐assisted selection.  相似文献   

16.
This study was conducted to investigate the immunomodulatory effect of a water‐soluble polysaccharide extracted from Artemisia argyi (AAP) in vitro. The effect was assessed in peripheral blood leucocytes (PBLs) of broilers, which were incubated with different AAP concentrations (0, 25, 50, 100, and 200 μg/ml) for 24 hr at 37°C in a 5% CO2 incubator. The results showed that, compared with the control group, immunoglobulin M (IgM) concentration was increased in the supernatant of the 100 μg/ml AAP‐treated group (p < .05), and immunoglobulin G (IgG) concentration was increased in the supernatant of the 200 μg/ml of AAP group (p < .05). In terms of cytokine production, production of interleukin‐1beta (IL‐1β), interleukin‐6 (IL‐6) and tumour necrosis factor‐alpha (TNF‐α) in the supernatant was enhanced in the AAP group in a dose‐dependent function, as well as enhanced mRNA expressions were showed in the cells (p < .05). The highest concentration of these three cytokines was observed in different AAP groups (IL‐1β for 25 μg/ml of AAP, IL‐6 for 100, and 200 μg/ml of AAP, and TNF‐α for 100 μg/ml of AAP respectively). The concentration of nitric oxide (NO) was increased when using AAP at the concentration of 100 μg/ml (p < .05) as compared to the control group. No significant effects on inducible nitric oxide synthase, Toll‐like receptor 4 (TLR4), myeloid differentiation factor 88 and nuclear factor Kappa B (NF‐κB) mRNA level were observed at each concentration of AAP. In conclusion, we found that AAP can specifically promote the production of immunoglobulins (IgM and IgG), cytokines (IL‐1β, IL‐6 and TNF‐α), as well as the NO concentration in vitro, but not through the activation of the TLR4/NF‐κB signalling pathway.  相似文献   

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The objective of the present study was to conduct an adipogenic evaluation of different roughage sources and feeding levels during ruminant adipocyte differentiation in vitro. Six wether sheep were divided into a timothy hay feeding group (TFG, n = 3) and an Italian ryegrass straw feeding group (IFG, n = 3). The sheep were fed high‐roughage (HR), medium roughage (MR) and low‐roughage (LR) diets in a one‐way layout design each over a 6‐day period. Sheep serum samples collected on the last day of each dietary treatment were added to an adipogenic induction medium for differentiation of preadipocytes derived from sheep subcutaneous adipose tissue. The cytoplasmic lipid accumulations in the TFG serum‐treated preadipocytes were significantly higher than those of the IFG‐serum treated preadipocytes on day 12. Messenger RNA expression of CCAAT/enhancer‐binding protein (C/EBP)‐α, C/EBP‐β, C/EBP‐δ, fatty‐acid‐binding protein (aP2) and stearoyl‐coenzyme A desaturase (SCD) were regulated by each serum treatment. This study shows that different roughage source diets and roughage‐to‐concentrate ratio diets can regulate adipocyte differentiation via ruminant blood composition.  相似文献   

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