Previous studies have suggested that polypeptides extracted from milk, soybean, fish, eggs, and meat possess potential anti-inflammatory effects. To date, few studies have reported the anti-inflammatory function of st...Previous studies have suggested that polypeptides extracted from milk, soybean, fish, eggs, and meat possess potential anti-inflammatory effects. To date, few studies have reported the anti-inflammatory function of sturgeon peptides and their underlying mechanisms are unknown. The current study was therefore to determine the anti-inflammatory potential of sturgeon peptides with lipopolysaccharide (LPS)-induced RAW264.7 inflammatory model. Pepsin hydrolysate (PeH) was purified by ultrafiltration and Sephadex G-15 gel filtration chromatography. PeH significantly reduced the inflammatory mediator (NO) and inflammatory cytokines (IL-6, TNF-α and IL-1β) expression in a dose-dependent manner. Moreover, the purified sturgeon peptide (F2) possessed strong antioxidant potential and effectively inhibited DPPH and ABTS free radicals. F2 significantly suppressed the expression of MAPK, IκBα, and NF-κB p65, indicating that F2 exerted anti-inflammatory influence by the inhibition of MAPK and NF-κB pathways.展开更多
B-3 exopolysaccharide is extracted from the Antarctic psychrophilic bacterium Psychrobacter sp. B-3. We have previously shown that it activates macrophages and affects their immunoregulatory activities. To determine w...B-3 exopolysaccharide is extracted from the Antarctic psychrophilic bacterium Psychrobacter sp. B-3. We have previously shown that it activates macrophages and affects their immunoregulatory activities. To determine what genes are affected during this process, we detected the genes differentially expressed in cells of RAW264.7 macrophages treated with B-3 exopolysaccharide by transcriptomic analysis. B-3 exopolysaccharide treatment caused differential expression of 420 genes, of which 178 were up-regulated and 242 were down-regulated. These genes were shown to be involved in many aspects of cell function, mainly metabolism and immunity. Genes were enriched in multiple immune-related pathways, and the most significantly enriched genes were involved in antigen processing and presentation pathways. The pathway in which differentially expressed genes were the most significantly enriched was the metabolic pathway; specifically, the expression of many metabolic enzyme genes was altered by B-3 exopolysaccharide treatment. Additionally, the genes involved in metabolisms of amino acids, carbohydrates, lipids and nucleotides, varied to certain degrees. B-3 exopolysaccharide, therefore, appears to directly affect the immune function of RAW264.7 macrophages as an immunostimulant, or to indirectly change intracellular metabolism. This is the first study to determine the effect of an Antarctic psychrophilic bacterial exopolysaccharide on RAW264.7 macrophages. Our findings provide an important reference for research into the regulation of macrophage immune function by different polysaccharides.展开更多
It has been observed that the consumption of litchi often causes symptoms characterized by itching or sore throat,gum swelling,oral cavity ulcers and even fever and inflammation,which significantly impair the quality ...It has been observed that the consumption of litchi often causes symptoms characterized by itching or sore throat,gum swelling,oral cavity ulcers and even fever and inflammation,which significantly impair the quality of life of a large population.Using the RAW264.7 cell line,a step-by-step strategy was used to screen for the components in litchi fruits that elicited adverse reactions.The adverse reaction fractions were identified by mass spectrometry and analyzed using the SMART program,and a sequence alignment of the homologous proteins was performed.MTT tests were used to determine the cytotoxicity of a litchi protein extract in RAW264.7 macrophages,and real-time PCR was applied to analyze the expression of inflammatory genes in the RAW264.7 cells treated with lipopolysaccharide or the litchi protein extract.The results showed that the litchi water-soluble protein extract could increase the production of the pro-inflammatory mediators IL-1β,iNOS and COX-2,and the anti-inflammatory mediator HO-1 in the RAW264.7 cell line.The 14-3-3-like proteins GF14 lambda,GF14 omega and GF14 upsilon were likely the candidate proteins that caused the adverse effects.展开更多
Xiaoli Wang,Xiaorong Hu,Huiqing Yan,Zhaocheng Ma and Xiuxin Deng.Horticulture Research(2017)4,17059;doi:10.1038/hortres.2017.59;Publishedonline25October2017.Correction to:Horticulture Research(2016)3,16017;doi:10.1038...Xiaoli Wang,Xiaorong Hu,Huiqing Yan,Zhaocheng Ma and Xiuxin Deng.Horticulture Research(2017)4,17059;doi:10.1038/hortres.2017.59;Publishedonline25October2017.Correction to:Horticulture Research(2016)3,16017;doi:10.1038/hortres.2016.17;Published online 04 May 2016.Since the publication of this article,the authors have noticed an error in Figure 6,the correct Figure 6 should be.展开更多
Aim Recent study have reported 2-Methoxy-6-acetyl-7-methyljuglone （MAM） for its potential antimicro- bial, neuroprotective and anticancer activity. However, the anti-inflammation effects of MAM remains to be elucida...Aim Recent study have reported 2-Methoxy-6-acetyl-7-methyljuglone （MAM） for its potential antimicro- bial, neuroprotective and anticancer activity. However, the anti-inflammation effects of MAM remains to be elucida- ted. We investigated the anti-inflammation activity of MAM. Methods RAW 264.7 macrophages were exposed to LPS with or without MAM. Inducible nitric oxide synthase （iNOS） expression and signaling molecules activated by LPS were evaluated. Results LPS-induced iNOS expression and nitric oxide （NO） expression was suppressed by MAM. MAM attenuated p38MAPK in cells treated with LPS. In addition, MAM caused an increase in MKP-1 expres- sion, which could suppress p38MAPK phosphorylation. Conclusions MAM may activate MKP-1, which then de- phosphorylates p38MAPK, resulting in iNOS down-regulation in LPS-stimulated RAW264.7 macrophages. The pres- ent study indicate that MAM may possess the potential to alleviate LPS-associated inflammatory disorders.展开更多
背景:溶质载体家族1成员5(solute carrier family 1 member 5,SLC1A5)在多种疾病中发挥了潜在作用,但确切作用机制尚不清楚。构建稳定的SLC1A5过表达和敲低细胞模型可为深入研究SLC1A5在疾病中的确切作用机制以及发现潜在治疗靶点提供...背景:溶质载体家族1成员5(solute carrier family 1 member 5,SLC1A5)在多种疾病中发挥了潜在作用,但确切作用机制尚不清楚。构建稳定的SLC1A5过表达和敲低细胞模型可为深入研究SLC1A5在疾病中的确切作用机制以及发现潜在治疗靶点提供有力的实验工具。目的:构建小鼠SLC1A5过表达和敲低的慢病毒载体,以建立稳定转染的RAW264.7细胞株,为深入探讨SLC1A5在炎症中的作用提供实验基础。方法:根据SLC1A5基因序列设计合成引物并使用聚合酶链反应扩增该基因片段。将目的基因定向接入经Age I/Nhe I酶切的载体质粒GV492中构建重组慢病毒质粒,对阳性克隆进一步筛选后测序比对结果;pHelper1.0质粒载体、pHelper2.0质粒载体、目的质粒载体与293T细胞共同培养并转染,获得慢病毒原液进行包装和滴度测定;在此基础上,通过体外培养RAW264.7细胞,确定嘌呤霉素工作质量浓度;不同滴度的慢病毒分别与RAW264.7细胞共同培养,根据荧光强度确定转染效率;用嘌呤霉素挑选出稳定转染细胞,实时荧光定量聚合酶链反应和蛋白免疫印迹方法检测稳定转染细胞株的SLC1A5基因和蛋白表达水平。结果与结论:(1)测序序列与目的序列一致提示重组慢病毒载体构建成功;(2)过表达SLC1A5慢病毒的滴度为1×10~9 TU/mL,敲低SLC1A5慢病毒的滴度为3×10~9 TU/mL;(3)确定RAW264.7细胞嘌呤霉素工作质量浓度为3μg/mL;(4)过表达/敲低SLC1A5慢病毒转染RAW264.7细胞的最佳条件皆为HiTransG P转染增强液且感染复数值等于50;(5)过表达SLC1A5稳转细胞株中SLC1A5基因和蛋白的表达量明显上调,而敲低SLC1A5稳转细胞株中SLC1A5基因和蛋白的表达量显著下调。结果表明,成功构建了小鼠SLC1A5过表达和敲低的慢病毒载体并获得稳定转染的RAW264.7细胞株。展开更多
Alkaloids are the main active constituents of the nearly ripe fruits of Euodia rutaecarpa(Juss.)Benth.,which possesses antitumor,anti-inflammatory,analgesic and antimicrobial activities.In this study,we aimed to asses...Alkaloids are the main active constituents of the nearly ripe fruits of Euodia rutaecarpa(Juss.)Benth.,which possesses antitumor,anti-inflammatory,analgesic and antimicrobial activities.In this study,we aimed to assess the potential anti-inflammatory effects of six main components,evodiamine,rutaecarpine,dehydroevodiamine,evocarpine,dihydroevocarpine and 1-methyl-2-undecyl-4-(1H)-quinolone,in the nearly ripe fruits of E.rutaecarpa.In the co-culture system consisting of the human colorectal adenocarcinoma(Caco-2)cells and RAW264.7 macrophages,inflammation was induced in RAW264.7 cells with 2μg/mL lipopolysaccharide.The results indicated that evodiamine,rutaecarpine,and dehydroevodiamine significantly down-regulated the mRNA expressions of pro-inflammatory cytokines(Il-6,Il-1βand Tnf-α)and inflammatory mediators,such as cyclooxygenase-2(Cox-2)and inducible nitric oxide synthase(iNos).Besides,the drug administration group showed a higher transepithelial electrical resistance and a lower tight junction permeability compared with the model group.Taken together,the tested compounds possessed anti-inflammatory effects.Our findings laid the foundation for further research on the anti-inflammatory mechanism of the nearly ripe fruits of E.rutaecarpa.展开更多
Objective:To explore the anti-inflammatory effect of the traditional Chinese medicine Zhikang capsule(ZKC)on lipopolysaccharide(LPS)-induced RAW264.7 cells.Methods:Safe concentrations of ZKC(0.175,0.35,and 0.7 mg/mL)w...Objective:To explore the anti-inflammatory effect of the traditional Chinese medicine Zhikang capsule(ZKC)on lipopolysaccharide(LPS)-induced RAW264.7 cells.Methods:Safe concentrations of ZKC(0.175,0.35,and 0.7 mg/mL)were used after the half-maximal inhibitory concentration(IC_(50))of RAW264.7 cells was calculated through the CCK-8 assay.In addition,the optimal intervention duration of ZKC(0.7 mg/mL)on RAW264.7 cells was determined to be 6 h,since all proinflammatory mediators[tumor necrosis factor-alpha(TNF-α),interleukin-1 beta(IL-1β),inteleukin-6(IL-6),cyclooxygenase-2(COX-2),inducible nitric oxide synthase(iNOS),and monocyte chemotactic protein-1(MCP-1)]had a decreasing tendency and relatively down-regulated mRNA expression levels as compared with other durations(4,8,and 12 h).RAW264.7 cells were pretreated with ZKC at various concentrations(0.175,0.35 and 0.7 mg/mL)for 6 h and then stimulated with LPS(1 μg/mL)for an additional 12 h.Results:In terms of inflammation,ZKC could reverse LPS-induced upregulation of TNF-α,IL-1β,IL-6,COX-2,iNOS,and MCP-1 at both the mRNA and protein levels in RAW264.7 cells in a dose-dependent manner.In terms of the NF-κB signaling pathway,ZKC could reduce phosphorylated p65 and promote M2 polarization of RAW264.7 cells under LPS stimulation in a dose-dependent manner.Moreover,ZKC exhibited a protective effect on macrophages from apoptosis.Conclusion:ZKC exhibited obvious antiinflammatory and anti-apoptotic effects on LPS-induced RAW264.7 cells at the cellular level,and a weakened NF-κB signaling pathway may be a potential significant target.展开更多
The anti-inflammatory activity of tea polyphenols(TPs)in RAW264.7 macrophages stimulated by lipopolysaccharide(LPS)was investigated in this paper.RAW264.7 macrophages were treated with different concentrations of TP(0...The anti-inflammatory activity of tea polyphenols(TPs)in RAW264.7 macrophages stimulated by lipopolysaccharide(LPS)was investigated in this paper.RAW264.7 macrophages were treated with different concentrations of TP(0,12.5,25,50,100,and 200μg/mL)and then stimulated by LPS.Another blank control group was set up.The expression of pro-inflammatory cytokines associated with the nuclear factor-kappa B(NF-κB)signaling pathway was investigated before and after TP treatment.Pretreatment of RAW264.7 cells with TP decreased the expression of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and interleukin 1 beta(IL-1β)pro-inflammatory cytokines.In addition,TP inhibited the phosphorylation of p65 and IκB by blocking the phosphorylation and the degradation of NF-κB inhibitor protein.In conclusion,TP exerts anti-inflammatory effects by regulating the release of inflammatory mediators via the NF-κB signaling pathway.展开更多
This study aimed to characterize the chemical composition of a new sulfated polysaccharide from the red alga Gracilaria chouae and evaluate its activation effects on RAW264.7 macrophages.It showed that the obtained G....This study aimed to characterize the chemical composition of a new sulfated polysaccharide from the red alga Gracilaria chouae and evaluate its activation effects on RAW264.7 macrophages.It showed that the obtained G.chouae polysaccharide(GCP-3A)was a sulfated acidic polysaccharide with a molecular weight of 11.87 kDa.展开更多
文摘Previous studies have suggested that polypeptides extracted from milk, soybean, fish, eggs, and meat possess potential anti-inflammatory effects. To date, few studies have reported the anti-inflammatory function of sturgeon peptides and their underlying mechanisms are unknown. The current study was therefore to determine the anti-inflammatory potential of sturgeon peptides with lipopolysaccharide (LPS)-induced RAW264.7 inflammatory model. Pepsin hydrolysate (PeH) was purified by ultrafiltration and Sephadex G-15 gel filtration chromatography. PeH significantly reduced the inflammatory mediator (NO) and inflammatory cytokines (IL-6, TNF-α and IL-1β) expression in a dose-dependent manner. Moreover, the purified sturgeon peptide (F2) possessed strong antioxidant potential and effectively inhibited DPPH and ABTS free radicals. F2 significantly suppressed the expression of MAPK, IκBα, and NF-κB p65, indicating that F2 exerted anti-inflammatory influence by the inhibition of MAPK and NF-κB pathways.
基金The Important National Science&Technology Specific Projects under contract No.2011ZX8001-003the National Natural Science Fundation of China under contract No.40706053Chinese Polar Environment Comprehensive Investigation&Assessment Programs under contract No.CHINARE2017-01-05
文摘B-3 exopolysaccharide is extracted from the Antarctic psychrophilic bacterium Psychrobacter sp. B-3. We have previously shown that it activates macrophages and affects their immunoregulatory activities. To determine what genes are affected during this process, we detected the genes differentially expressed in cells of RAW264.7 macrophages treated with B-3 exopolysaccharide by transcriptomic analysis. B-3 exopolysaccharide treatment caused differential expression of 420 genes, of which 178 were up-regulated and 242 were down-regulated. These genes were shown to be involved in many aspects of cell function, mainly metabolism and immunity. Genes were enriched in multiple immune-related pathways, and the most significantly enriched genes were involved in antigen processing and presentation pathways. The pathway in which differentially expressed genes were the most significantly enriched was the metabolic pathway; specifically, the expression of many metabolic enzyme genes was altered by B-3 exopolysaccharide treatment. Additionally, the genes involved in metabolisms of amino acids, carbohydrates, lipids and nucleotides, varied to certain degrees. B-3 exopolysaccharide, therefore, appears to directly affect the immune function of RAW264.7 macrophages as an immunostimulant, or to indirectly change intracellular metabolism. This is the first study to determine the effect of an Antarctic psychrophilic bacterial exopolysaccharide on RAW264.7 macrophages. Our findings provide an important reference for research into the regulation of macrophage immune function by different polysaccharides.
基金This work was supported by the Special Fund for Agro-scientific Research in the Public Interest(No.201403036)the Fundamental Research Funds for the Central Universities(2014PY030)+1 种基金the National Basic Research Program of China(2011CB100600)the National NSF of China and the Foundation for the Key Program of the Ministry of Education,China(311029).
文摘It has been observed that the consumption of litchi often causes symptoms characterized by itching or sore throat,gum swelling,oral cavity ulcers and even fever and inflammation,which significantly impair the quality of life of a large population.Using the RAW264.7 cell line,a step-by-step strategy was used to screen for the components in litchi fruits that elicited adverse reactions.The adverse reaction fractions were identified by mass spectrometry and analyzed using the SMART program,and a sequence alignment of the homologous proteins was performed.MTT tests were used to determine the cytotoxicity of a litchi protein extract in RAW264.7 macrophages,and real-time PCR was applied to analyze the expression of inflammatory genes in the RAW264.7 cells treated with lipopolysaccharide or the litchi protein extract.The results showed that the litchi water-soluble protein extract could increase the production of the pro-inflammatory mediators IL-1β,iNOS and COX-2,and the anti-inflammatory mediator HO-1 in the RAW264.7 cell line.The 14-3-3-like proteins GF14 lambda,GF14 omega and GF14 upsilon were likely the candidate proteins that caused the adverse effects.
文摘Xiaoli Wang,Xiaorong Hu,Huiqing Yan,Zhaocheng Ma and Xiuxin Deng.Horticulture Research(2017)4,17059;doi:10.1038/hortres.2017.59;Publishedonline25October2017.Correction to:Horticulture Research(2016)3,16017;doi:10.1038/hortres.2016.17;Published online 04 May 2016.Since the publication of this article,the authors have noticed an error in Figure 6,the correct Figure 6 should be.
文摘Aim Recent study have reported 2-Methoxy-6-acetyl-7-methyljuglone （MAM） for its potential antimicro- bial, neuroprotective and anticancer activity. However, the anti-inflammation effects of MAM remains to be elucida- ted. We investigated the anti-inflammation activity of MAM. Methods RAW 264.7 macrophages were exposed to LPS with or without MAM. Inducible nitric oxide synthase （iNOS） expression and signaling molecules activated by LPS were evaluated. Results LPS-induced iNOS expression and nitric oxide （NO） expression was suppressed by MAM. MAM attenuated p38MAPK in cells treated with LPS. In addition, MAM caused an increase in MKP-1 expres- sion, which could suppress p38MAPK phosphorylation. Conclusions MAM may activate MKP-1, which then de- phosphorylates p38MAPK, resulting in iNOS down-regulation in LPS-stimulated RAW264.7 macrophages. The pres- ent study indicate that MAM may possess the potential to alleviate LPS-associated inflammatory disorders.
基金National Key R&D Program of China(Grant No.2018YFC1704500,2018YFC1704506)the National Natural Science Foundation of China(Grant No.81773865)。
文摘Alkaloids are the main active constituents of the nearly ripe fruits of Euodia rutaecarpa(Juss.)Benth.,which possesses antitumor,anti-inflammatory,analgesic and antimicrobial activities.In this study,we aimed to assess the potential anti-inflammatory effects of six main components,evodiamine,rutaecarpine,dehydroevodiamine,evocarpine,dihydroevocarpine and 1-methyl-2-undecyl-4-(1H)-quinolone,in the nearly ripe fruits of E.rutaecarpa.In the co-culture system consisting of the human colorectal adenocarcinoma(Caco-2)cells and RAW264.7 macrophages,inflammation was induced in RAW264.7 cells with 2μg/mL lipopolysaccharide.The results indicated that evodiamine,rutaecarpine,and dehydroevodiamine significantly down-regulated the mRNA expressions of pro-inflammatory cytokines(Il-6,Il-1βand Tnf-α)and inflammatory mediators,such as cyclooxygenase-2(Cox-2)and inducible nitric oxide synthase(iNos).Besides,the drug administration group showed a higher transepithelial electrical resistance and a lower tight junction permeability compared with the model group.Taken together,the tested compounds possessed anti-inflammatory effects.Our findings laid the foundation for further research on the anti-inflammatory mechanism of the nearly ripe fruits of E.rutaecarpa.
文摘Objective:To explore the anti-inflammatory effect of the traditional Chinese medicine Zhikang capsule(ZKC)on lipopolysaccharide(LPS)-induced RAW264.7 cells.Methods:Safe concentrations of ZKC(0.175,0.35,and 0.7 mg/mL)were used after the half-maximal inhibitory concentration(IC_(50))of RAW264.7 cells was calculated through the CCK-8 assay.In addition,the optimal intervention duration of ZKC(0.7 mg/mL)on RAW264.7 cells was determined to be 6 h,since all proinflammatory mediators[tumor necrosis factor-alpha(TNF-α),interleukin-1 beta(IL-1β),inteleukin-6(IL-6),cyclooxygenase-2(COX-2),inducible nitric oxide synthase(iNOS),and monocyte chemotactic protein-1(MCP-1)]had a decreasing tendency and relatively down-regulated mRNA expression levels as compared with other durations(4,8,and 12 h).RAW264.7 cells were pretreated with ZKC at various concentrations(0.175,0.35 and 0.7 mg/mL)for 6 h and then stimulated with LPS(1 μg/mL)for an additional 12 h.Results:In terms of inflammation,ZKC could reverse LPS-induced upregulation of TNF-α,IL-1β,IL-6,COX-2,iNOS,and MCP-1 at both the mRNA and protein levels in RAW264.7 cells in a dose-dependent manner.In terms of the NF-κB signaling pathway,ZKC could reduce phosphorylated p65 and promote M2 polarization of RAW264.7 cells under LPS stimulation in a dose-dependent manner.Moreover,ZKC exhibited a protective effect on macrophages from apoptosis.Conclusion:ZKC exhibited obvious antiinflammatory and anti-apoptotic effects on LPS-induced RAW264.7 cells at the cellular level,and a weakened NF-κB signaling pathway may be a potential significant target.
文摘The anti-inflammatory activity of tea polyphenols(TPs)in RAW264.7 macrophages stimulated by lipopolysaccharide(LPS)was investigated in this paper.RAW264.7 macrophages were treated with different concentrations of TP(0,12.5,25,50,100,and 200μg/mL)and then stimulated by LPS.Another blank control group was set up.The expression of pro-inflammatory cytokines associated with the nuclear factor-kappa B(NF-κB)signaling pathway was investigated before and after TP treatment.Pretreatment of RAW264.7 cells with TP decreased the expression of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and interleukin 1 beta(IL-1β)pro-inflammatory cytokines.In addition,TP inhibited the phosphorylation of p65 and IκB by blocking the phosphorylation and the degradation of NF-κB inhibitor protein.In conclusion,TP exerts anti-inflammatory effects by regulating the release of inflammatory mediators via the NF-κB signaling pathway.
基金This work was supported by the National Natural Science Foundation of China(No.81572989)the International Academic Cooperation of Science and Technology Committee of Shanghai,China(No.18430721100).
文摘This study aimed to characterize the chemical composition of a new sulfated polysaccharide from the red alga Gracilaria chouae and evaluate its activation effects on RAW264.7 macrophages.It showed that the obtained G.chouae polysaccharide(GCP-3A)was a sulfated acidic polysaccharide with a molecular weight of 11.87 kDa.
