Ting Yi; Zhiyin Liu; Haokun Jia; Qiongzhi Liu; Jianjiao Peng
Abstract
Background: The imbalance between M1 and M2 macrophage activation is closely associated with the pathogenesis of inflammatory bowel diseases (IBDs). Sulforaphane (SFN) plays an important role in the treatment of inflammatory diseases.Objective: To investigate the effect of SFN on macrophage polarization ...
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Background: The imbalance between M1 and M2 macrophage activation is closely associated with the pathogenesis of inflammatory bowel diseases (IBDs). Sulforaphane (SFN) plays an important role in the treatment of inflammatory diseases.Objective: To investigate the effect of SFN on macrophage polarization and its underlying regulatory mechanism.Methods: Mouse bone marrow-derived macrophages (BMDMs) were treated with SFN and an Nrf2 inhibitor, Brusatol. M1 macrophages were induced by LPS and IFN-γ stimulation, whereas M2 macrophages were induced by stimulation with IL-4 and IL-13. LPS-stimulated BMDMs were co-cultured with Caco-2 cells. Flow cytometry, qRT-PCR, and Western blot were performed to assess macrophage polarization. Cell function was assessed using CCK8 assay, transepithelial electrical resistance (TEER) assay, and biochemical analysis.Results: Higher concentrations of SFN resulted in better intervention effects, with an optimal concentration of 10 μM. SFN decreased the levels of IL-12, IL-6, and TNF-α, as well as the percentages of CD16/32 in M1 BMDMs. At the same time, SFN increased the levels of YM1, Fizz1, and Arg1 as well as the percentages of CD206+ cells in M2 BMDMs. In addition, SFN enhanced the accumulation of Nrf2, NQO1, and HO-1 in M1 BMDMs, and the downregulation of Nrf2 reversed the regulatory effect of SFN on M1/M2 macrophages. LPS-stimulated BMDMs induced Caco-2 cell damage, which was partially alleviated by SFN.Conclusion: Our findings indicate that SFN may act as an Nrf2 agonist to regulate macrophage polarization from M1 to M2. Furthermore, SFN may represent a potential protective ingredient against IBD.
Enna Liu; Zheng Li; Yan Zhang; Kuisheng Chen
Abstract
Background: Macrophage polarization plays a critical role in determining the inflammatory states. Hepcidin is a key negative regulator of iron homeostasis and functions. Although hepcidin has been shown to affect ferroportin expression in macrophages, whether it affects macrophage polarization is still ...
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Background: Macrophage polarization plays a critical role in determining the inflammatory states. Hepcidin is a key negative regulator of iron homeostasis and functions. Although hepcidin has been shown to affect ferroportin expression in macrophages, whether it affects macrophage polarization is still largely unknown. Objective: To address whether hepcidin induces macrophage polarization. Methods: The expression of iNOS and CD206, and the ratio of IFN-γ vs IL-4 in THP-1 derived macrophages upon hepcidin stimulation were evaluated. Further detected was the percentage of CD16+ M1, CD23+ M1, CD10+ M2 and CCL22+ M2 cells in monocyte derived macrophages. Results: M1 associated molecules were increased in hepcidin-treated cells, yet M2 associated molecules were increased when hepcidin was neutralized. Concomitantly, we observed a significant increase in IRF3 phosphorylation in hepcidin-stimulated cells. However, STAT6 phosphorylation with hepcidin was neutralized. Conclusion: Hepcidin is able to induce macrophage polarization towards M1 type, and might be utilized as a potential M1 macrophage agonist in clinical practice.
Gege Li; Jiahui Pan; Qiuling Tang; Xinchan Liu; Liuran Wang; Yang Meng; Weixian Yu
Abstract
Background: C5areceptor antagonistPMX205 is a synthetic hexapeptidecapable of blocking C5a-C5a receptor (C5aR) axis by simulating C5a active C-terminal amino acid residues. This hexapeptide presents good anti-inflammatory effects in a myriad inflammation models. The anti-inflammatory effect of PMX205 ...
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Background: C5areceptor antagonistPMX205 is a synthetic hexapeptidecapable of blocking C5a-C5a receptor (C5aR) axis by simulating C5a active C-terminal amino acid residues. This hexapeptide presents good anti-inflammatory effects in a myriad inflammation models. The anti-inflammatory effect of PMX205 on periodontitis is yet to be fully fathomed. Objective: To examine the anti-inflammatory effects of PMX205 on RAW264.7 murine macrophages exposed togingipain extracts and Porphyromonas gingivalis (P. gingivalis). Methods: MTT assay was carried out so as to specify the cytotoxicity of PMX205. RAW264.7 cells were co-cultured in vitro with gingipain extracts or P. gingivalis to simulate the periodontitis inflammatory milieu. Real-time quantitative PCR, ELISA and Griess assay were performed in order to detect tumor necrosis factor-α (TNF-α), IL-6, IL-23, nitric oxide (NO), IL-10, transforming growth factor-β1 (TGF-β1), andarginase-1 (Arg-1). Furthermore, phagocytosis assay was done to evaluate the phagocytic capacity of RAW 264.7 cells. Finally, western blot analysis was conducted to evaluate myeloid differentiation factor 88 (MyD88). Results: PMX205 increased the expression levels of bacteriostatic substances (NO and IL-23) and anti-inflammatory cytokines (TGF-β1, IL-10 and Arg-1); however, it reduced the expression levels of proinflammatory cytokines TNF-α and IL-6once RAW 264.7 macrophages were stimulated via gingipain extracts or P. gingivalis. In addition, PMX205 promoted the macrophage phagocytosis and down-regulated protein expression of MyD88. Conclusion: PMX205 has recognizable anti-inflammatory effects in RAW 264.7 cell inflammation model, a finding which probably opens doors to future investigations on new targets for the prevention and treatment of chronic periodontitis.
Shahid Waseem; Kashif-Ur-Rehman -; Ramesh Kumar; Tariq Mahmood
Volume 13, Issue 1 , March 2016, , Pages 1-8
Abstract
Background: Falciparum malaria is a severe health burden worldwide. Antigen presenting cells are reported to be affected by erythrocytic stage of the parasite. Malarial hemozoin (HZ), a metabolite of malaria parasite, has adjuvant properties and may play a role in the induction of immune response against ...
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Background: Falciparum malaria is a severe health burden worldwide. Antigen presenting cells are reported to be affected by erythrocytic stage of the parasite. Malarial hemozoin (HZ), a metabolite of malaria parasite, has adjuvant properties and may play a role in the induction of immune response against the parasite. Objective: To determine the immunological impact of hemozoin on the capacity of innate immune cells maturation. Methods: Plasmodium falciparum (F32 strain) was cultured in O+ blood group up to 18% parasitemia. Natural hemozoin was extracted from infected red blood cells. Murine bone marrow derived macrophages and myeloid dendritic cells were stimulated with 4 ߤg/mL or 40 ߤg/mL of synthetic hemozoin (β-hematin) or natural hemozoin. We assessed the immunomodulatory role of synthetic or natural hemozoin in vitro by flowcytometric analysis. Results: The maturation markers MHCII, CD80 and CD86 were significantly upregulated (p<0.05) on the surface of murine bone marrow derived macrophages or myeloid dendritic cells. Data confirmed the potential of macrophages or myeloid dendritic cells, through hemozoin activation, to establish an innate immune response against malaria parasites. Conclusion: Both synthetic and natural hemozoin are potent inducers of cellular immunity against malaria infection. However, natural hemozoin is a stronger inducer as compared to synthetic hemozoin.
Ali Shams Shahemabadi; Ahmad Zavaran Hosseini; Shapour Shaghasempour; Mohammad Reza Masjedi; Majid Rayani; Majid Shams; Nasrin Esphandyari; Majid Pouramiri
Volume 7, Issue 1 , March 2010, , Pages 57-63
Abstract
Background: Mycobacterium tuberculosis lipid antigens take part in pathogenicity of the bacterium but the response of monocytes/macrophages to these antigens in tubercu-losis is not well known. Objective: The aim of current investigation was to study the M. tuberculosis lipid antigens in tuberculosis ...
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Background: Mycobacterium tuberculosis lipid antigens take part in pathogenicity of the bacterium but the response of monocytes/macrophages to these antigens in tubercu-losis is not well known. Objective: The aim of current investigation was to study the M. tuberculosis lipid antigens in tuberculosis pathogenesis. Methods: In the present study M. tuberculosis lipid antigens were extracted. Monocytes and macrophages from mul-tidrug-resistant tuberculosis (MDR-TB), TB patients, asymptomatic healthy individuals with positive tuberculin skin test positive and healthy individuals with negative tubercu-lin skin test were collected using magnetic cell sorting. The cells were stimulated by M. tuberculosis total lipid antigens and IL-12 and IL-10 in their supernatants were meas-ured by enzyme-linked immunosorbent assay. Results: The IL-12 production by mono-cytes in response to M. tuberculosis total sonicate antigens in the MDR-TB patients did not show a considerable difference with the PPD positive healthy subjects, whereas in the active TB patients, IL-12 levels significantly decreased (p<0.05). IL-10 production by monocytes in TB patients in response to total lipid antigens showed a significant in-crease in comparison to MDR-TB patients and healthy individuals. Conclusion: In the MDR-TB patients, IL-10 and IL-12 production by monocytes in response to M. tubercu-losis lipid antigens are similar to the healthy subjects.
Kazem Ahmadi; Majid Riazipour
Volume 4, Issue 4 , December 2007, , Pages 220-226
Abstract
Background: The water-soluble extract of Ganoderma lucidum (Reishi) has been used as an immunomodulator to stimulate spleen cells proliferation and cytokine expression. Objective: To investigate the effect of Ganoderma lucidum (G. lucidum) on cytokine production by mice peritoneal macrophages. Methods: ...
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Background: The water-soluble extract of Ganoderma lucidum (Reishi) has been used as an immunomodulator to stimulate spleen cells proliferation and cytokine expression. Objective: To investigate the effect of Ganoderma lucidum (G. lucidum) on cytokine production by mice peritoneal macrophages. Methods: Mice peritoneal macrophages were prepared by intra-peritoneal injection of 5 ml cold PBS. Peritoneal macrophages were plated out at 1X106 cell/well in 1ml RPMI 1640 medium supplemented with 10%FCS, 50 μg streptomycin and 50U penicillin. Cells were incubated in the presence or absence of different concentrations of G. lucidum at 370C and 5% CO2 for 48 hours. Cell free medium was removed and used for cytokine assay by ELISA method (Bender med system). Results: The results showed no significant differences in cell viability at concentrations ranged from 0-40 μg/ml compared with control group. G. lucidum enhanced IL-1β, TNF-α and NO production in a concentration dependent manner. However, it is not clear if the enhancement of NO release is due to direct effect of G. lucidum on NO synthesis or by indirect endogenous modulation via cytokines. IL-12 release by peritoneal macrophages was also increased in response to different concentrations of G. lucidum, but maximum enhancement was induced in response to 5 μg/ml of G. lucidum (p<0.001). Conclusion: Our results indicate that G. lucidum at concentrations used has a positive effect on cytokine release and NO production by peritoneal macrophages. Therefore, it is concluded that G. lucidum at moderate concentrations improves macrophage function through cytokine and NO release.
