scholarly journals The effect of the WKYMVm peptide on promoting mBMSC secretion of exosomes to induce M2 macrophage polarization through the FPR2 pathway

2021 ◽  
Vol 16 (1) ◽  
Author(s):  
Wenbo Zhao ◽  
Junxian Hu ◽  
Qingyi He
Keyword(s):  
Western Blotting ◽  
Macrophage Polarization ◽  
Bone Defects ◽  
The Other ◽  
Formyl Peptide Receptor ◽  
Cell Counting ◽  
M2 Macrophage ◽  
M2 Macrophages ◽  
Lysosomal Activity ◽  
M2 Macrophage Polarization

Abstract Background When multicystic vesicles (precursors of exosomes) are formed in cells, there are two results. One is decomposition by lysosomes, and the other is the generation of exosomes that are transported out through the transmembrane. On the other hand, M2 macrophages promote the formation of local vascularization and provide necessary support for the repair of bone defects. To provide a new idea for the treatment of bone defects, the purpose of our study was to investigate the effect of WKYMVm (Trp-Lys-Tyr-Met-Val-D-Met-NH2) peptide on the secretion of exosomes from murine bone marrow-derived MSCs (mBMSCs) and the effect of exosomes on the polarization of M2 macrophages. Methods The WKYMVm peptide was used to activate the formyl peptide receptor 2 (FPR2) pathway in mBMSCs. First, we used Cell Counting Kit-8 (CCK-8) to detect the cytotoxic effect of WKYMVm peptide on mBMSCs. Second, we used western blotting (WB) and quantitative real-time polymerase chain reaction (qRT-PCR) to detect the expression of interferon stimulated gene 15 (ISG15) and transcription factor EB (TFEB) in mBMSCs. Then, we detected lysosomal activity using a lysozyme activity assay kit. Third, we used an exosome extraction kit and western blotting to detect the content of exosomes secreted by mBMSCs. Fourth, we used immunofluorescence and western blotting to count the number of polarized M2 macrophages. Finally, we used an inhibitor to block miRNA-146 in exosomes secreted by mBMSCs and counted the number of polarized M2 macrophages. Results We first found that the WKYMVm peptide had no toxic effect on mBMSCs at a concentration of 1 μmol/L. Second, we found that when the FPR2 pathway was activated by the WKYMVm peptide in mBMSCs, ISG15 and TFEB expression was decreased, leading to increased secretion of exosomes. We also found that lysosomal activity was decreased when the FPR2 pathway was activated by the WKYMVm peptide in mBMSCs. Third, we demonstrated that exosomes secreted by mBMSCs promote the polarization of M2 macrophages. Moreover, all these effects can be blocked by the WRWWWW (WRW4, H-Trp-Arg-Trp-Trp-Trp-Trp-OH) peptide, an inhibitor of the FPR2 pathway. Finally, we confirmed the effect of miRNA-146 in exosomes secreted by mBMSCs on promoting the polarization of M2 macrophages. Conclusion Our findings demonstrated the potential value of the WKYMVm peptide in promoting the secretion of exosomes by mBMSCs and eventually leading to M2 macrophage polarization. We believe that our study could provide a research basis for the clinical treatment of bone defects.

scholarly journals The Effect of WKYMVm Peptide on Promoting mBMSCs Secrete Exosomes to Make M2 Macrophages Polarization Through FPR2 Pathway

2020 ◽  
Author(s):  
Wenbo Zhao ◽  
Junxian Hu ◽  
QingYi He
Keyword(s):  
Western Blotting ◽  
Bone Defects ◽  
The Other ◽  
Formyl Peptide Receptor ◽  
M2 Macrophages ◽  
Murine Bone Marrow ◽  
Macrophages Polarization ◽  
Formyl Peptide ◽  
Transcription Factor Eb ◽  
Interferon Stimulated Gene 15

Abstract Background: When the multicystic vesicles (precursor of exosomes) are formed in cells, there are two results. One is to be decomposed by lysosomes and the other one is to become exosomes which are transported out through transmembrane. On the other hand, M2 macrophages can promote the formation of local vascularization and provide necessary support for the repair of bone defects. In order to provide a new idea for the treatment of bone defects, the purpose of our study is to investigate the effect of WKYMVm peptide on the secretion of exosomes from murine bone marrow-derived MSCs （mBMSCs） and the effect of exosomes on the polarization of M2 macrophages. Methods: WKYMVm peptide was used to activate the Formyl Peptide Receptor 2 (FPR2) pathway in mBMSCs. At first, we used CCK-8 kit to detect the cytotoxic effect of WKYMVm Peptide on mBMSCs. Secondly, we used western blotting and Quantitative real-time Polymerase Chain Reaction （Qrt-PCR） to detect the expression of Interferon Stimulated Gene 15 （ISG15） and Transcription Factor EB （TFEB） in mBMSCs. Thirdly, we used exosomes extraction kit and western blotting to detect the content of exosomes secreted by mBMSCs. Finally, we used the immunofluorescence and western blotting to count the number of M2 macrophages polarization.Results: We firstly found that WKYMVm peptide had no toxic effect on mBMSCs at the concentration of 1μmol/L. Secondly, we found that when the FPR2 pathway was activated by WKYMVm peptide in mBMSCs, the expression of ISG15 and TFEB were decreased, leaded to the secretion of exosomes increased. Finally, we proved that the exosomes secreted by mBMSCs could promote the polarization of M2 macrophages. Moreover, all these effects can be blocked by WRWWWW (WRW4) peptide, an inhibitor of FPR2 pathway.Conclusion: Our findings demonstrated the potential value of WKYMVm peptide in promoting the secretion of exosomes by mBMSCs and eventually leading to M2 macrophages polarization. We believed that our study could provide a research basis for the clinical treatment of bone defects.

scholarly journals ILC2 Proliferated by IL-33 Stimulation Alleviates Acute Colitis in Rag1-/- Mouse through Promoting M2 Macrophage Polarization

2020 ◽  
Vol 2020 ◽  
pp. 1-13
Author(s):  
Yong You ◽  
Xiaoqing Zhang ◽  
Xiao Wang ◽  
Dan Yue ◽  
Fanxiang Meng ◽  
...  
Keyword(s):  
Body Weight ◽  
Macrophage Polarization ◽  
Epithelial Barrier ◽  
M2 Macrophage ◽  
Surface Markers ◽  
M2 Macrophages ◽  
Immunological Function ◽  
Innate Lymphoid Cell ◽  
Acute Colitis ◽  
M2 Macrophage Polarization

This study was to identify functions of ILC2, a newly found innate lymphoid cell which mainly locates in mucosa organs like lungs and intestines, in IBD. We injected rIL-33 protein to C57/BL6 mouse to explore how IL-33 induces ILC2 proliferation. The results showed that ILC2 reached a proliferation peak at day 5 and expressed multiple surface markers like CD127, C-kit, CD69, CD44, ST2, CD27, DR3, MHCII, and CD90.2. ILC2 also expressed high quantity of IL-13 and IL-5 and few IL-17A which indicates a potentially immunological function in IBD development. Afterwards, we transferred sort purified ILC2 to Rag1-/- mouse given DSS to induce acute colitis in order to explore the innate function of ILC2. Data showed that ILC2 alleviates DSS-induced acute innate colitis by repairing epithelial barrier and restore body weight. Furthermore, we found that ILC2 can cause macrophages polarizing to M2 macrophages in the gut. Therefore, we concluded that ILC2 played a therapeutic role in mouse acute colitis.

scholarly journals Lung Tumor Cell-Derived Exosomes Promote M2 Macrophage Polarization

Cells ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 1303 ◽  
Author(s):  
Alexandra Pritchard ◽  
Sultan Tousif ◽  
Yong Wang ◽  
Kenneth Hough ◽  
Saad Khan ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Tumor Cells ◽  
Bone Marrow Cells ◽  
Lung Tumor ◽  
Macrophage Polarization ◽  
M2 Macrophage ◽  
M2 Macrophages ◽  
M2 Macrophage Polarization ◽  
The Impact ◽  
M2 Phenotype

Cellular cross-talk within the tumor microenvironment (TME) by exosomes is known to promote tumor progression. Tumor promoting macrophages with an M2 phenotype are suppressors of anti-tumor immunity. However, the impact of tumor-derived exosomes in modulating macrophage polarization in the lung TME is largely unknown. Herein, we investigated if lung tumor-derived exosomes alter transcriptional and bioenergetic signatures of M0 macrophages and polarize them to an M2 phenotype. The concentration of exosomes produced by p53 null H358 lung tumor cells was significantly reduced compared to A549 (p53 wild-type) lung tumor cells, consistent with p53-mediated regulation of exosome production. In co-culture studies, M0 macrophages internalized tumor-derived exosomes, and differentiated into M2 phenotype. Importantly, we demonstrate that tumor-derived exosomes enhance the oxygen consumption rate of macrophages, altering their bioenergetic state consistent with that of M2 macrophages. In vitro co-cultures of M0 macrophages with H358 exosomes demonstrated that exosome-induced M2 polarization may be p53 independent. Murine bone marrow cells and bone marrow-derived myeloid-derived suppressor cells (MDSCs) co-cultured with lewis lung carcinoma (LLC)-derived exosomes differentiated to M2 macrophages. Collectively, these studies provide evidence for a novel role for lung tumor-exosomes in M2 macrophage polarization, which then offers new therapeutic targets for immunotherapy of lung cancer.

scholarly journals Montelukast, a Cysteinyl Leukotriene Receptor 1 Antagonist, Induces M2 Macrophage Polarization and Inhibits Murine Aortic Aneurysm Formation

2019 ◽  
Vol 2019 ◽  
pp. 1-11 ◽  
Author(s):  
Yohei Kawai ◽  
Yuji Narita ◽  
Aika Yamawaki-Ogata ◽  
Akihiko Usui ◽  
Kimihiro Komori
Keyword(s):  
Aortic Aneurysm ◽  
Macrophage Polarization ◽  
Enzyme Linked Immunosorbent Assay ◽  
M2 Macrophage ◽  
M2 Macrophages ◽  
Gene Expressions ◽  
Arginase 1 ◽  
M2 Macrophage Polarization

Background. The pathogenesis of abdominal aortic aneurysm (AAA) is characterized by atherosclerosis with chronic inflammation in the aortic wall. Montelukast is a selective cys-LT 1 receptor antagonist that can suppress atherosclerotic diseases. We evaluated the in vitro properties of montelukast and its in vivo activities in an angiotensin II–infused apolipoprotein E–deficient (apoE−/−) AAA mouse model. Methods. The mouse monocyte/macrophage cell line J774A.1 was used in vitro. M1 macrophages were treated with montelukast, and gene expressions of inflammatory cytokines were measured. Macrophages were cultured with montelukast, then gene expressions of arginase-1 and IL (interleukin)-10 were assessed by quantitative polymerase chain reaction, arginase-1 was measured by fluorescence-activated cell sorting, and IL-10 concentration was analyzed by enzyme-linked immunosorbent assay. In vivo, one group (Mont, n=7) received oral montelukast (10 mg/kg/day) for 28 days, and the other group (Saline, n=7) was given normal Saline as a control for the same period. Aortic diameters, activities of matrix metalloproteinases (MMPs), cytokine concentrations, and the number of M2 macrophages were analyzed. Results. Relative to control, montelukast significantly suppressed gene expressions of MMP-2, MMP-9, and IL-1β, induced gene expressions of arginase-1 and IL-10, enhanced the expression of the arginase-1 cell surface protein, and increased the protein concentration of IL-10. In vivo, montelukast significantly decreased aortic expansion (Saline vs Mont; 2.44 ± 0.15 mm vs 1.59 ± 0.20 mm, P<.01), reduced MMP-2 activity (Saline vs Mont; 1240 μM vs 755 μM, P<.05), and induced infiltration of M2 macrophages (Saline vs Mont; 7.51 % vs 14.7 %, P<.05). Conclusion. Montelukast induces M2 macrophage polarization and prevents AAA formation in apoE−/− mice.

FPR2 Participates in Epithelial Ovarian Cancer (EOC) Progression Through RhoA Mediated M2 Macrophage Polarization: An Invitro Experimental Study

2020 ◽  
Author(s):  
Xiaohui Xie ◽  
Juan He ◽  
Yaqiong Liu ◽  
Weiwei Chen ◽  
Kun Shi
Keyword(s):  
Ovarian Cancer ◽  
Western Blot ◽  
Cell Lines ◽  
Macrophage Polarization ◽  
Ectopic Expression ◽  
Ovarian Cancer Cells ◽  
M2 Macrophage ◽  
M2 Macrophages ◽  
Invasion And Metastasis ◽  
M2 Macrophage Polarization

Abstract Background: In our previous study, we found Formyl peptide receptor 2 (FPR2) promoted the invasion and metastasis of EOC and it could be a prognostic marker for EOC. In this study, we aimed to study the possible mechanism of FPR2 in promoting EOC progression.Methods: The FPR2 ectopic expression and knockdown EOC cell lines as well as their control cell lines were established and the expression change of RhoA in each cell lines was evaluated by RT-qPCR and Western-blot. Wound healing and Transwell assays were performed to detect the migrational ability of EOCs that affected by FPR2 and RhoA. The supernatant of each EOC cell lines were used to co-culture with the macrophages, and tested the M1 and M2 macrophges biomarkers by flow cytometry. THP-1 cell line was also indcued to differentiated to M1 and M2 macrophages, FPR2 and RhoA expression in each macrophage cell lines were detected by RT-qPCR and Western-blot. Results: RhoA expression was significantly increased in EOCs along with the overexpression of FPR2, which showed a positive correlation by Pearson correlation analysis. FPR2 ectopic expression would contribute to the migrational ability of EOCs, and RhoA inhibitor (C3 transferase) would impare EOCs migration. Furthermore, FPR2 stimulated the secretion of Th2 cytokines by EOCs, which induced macrophages differentiate to M2 phenotype, while RhoA inhibitor stimulate the secretion of Th1 cytokines and induce macrophages differentiate to M1 phenotype. Moreover, compared with M1 macrophages and THP-1 cells, FPR2 and RhoA expression were significantly up-regulated in M2 macrophages.Conclusion: FPR2 stimulated M2 macrophage polarization and promote invasion and metastasis of ovarian cancer cells through RhoA.

scholarly journals Hexapeptide induces M2 macrophage polarization via the JAK1/STAT6 pathway to promote angiogenesis in bone repair

2021 ◽  
Author(s):  
Xinyun Han ◽  
Junxian Hu ◽  
Wenbo Zhao ◽  
Hongwei Lu ◽  
Jingjin Dai ◽  
...  
Keyword(s):  
Bone Repair ◽  
Macrophage Polarization ◽  
M2 Macrophage ◽  
M2 Macrophages ◽  
Promising Alternative ◽  
Defect Repair ◽  
M2 Macrophage Polarization ◽  
Underlying Mechanisms ◽  
Main Regulator

Abstract Angiogenesis is essential for successful bone defect repair. In normal tissue repair, the physiological inflammatory response is the main regulator of angiogenesis through the activity of macrophages and the cytokines secreted by them. In particular, M2 macrophages which secrete high levels of PDGF-BB are typically considered to promote angiogenesis. A hexapeptide [WKYMVm, (Trp-Lys-Tyr-Met-Val-D-Met-NH2)] has been reported to modulate inflammatory activities. However, the underlying mechanisms by which WKYMVm regulates macrophages remain unclear. In this study, the possible involvement by which WKYMVm induces the polarization of macrophages and affects their behaviors was evaluated. In vitro results showed that macrophages were induced to an M2 rather than M1 phenotype and the M2 phenotype was enhanced by WKYMVm through activation of the JAK1/STAT6 signaling pathway. It was also found that WKYMVm played an important role in the PDGF-BB production increase and proangiogenic abilities in M2 macrophages. Consistent with the results in vitro, the elevated M2/M0 ratio induced by WKYMVm enhanced the formation of new blood vessels in a femoral defect mouse model. In summary, these findings suggest that WKYMVm could be a promising alternative strategy for angiogenesis in bone repair by inducing M2 macrophage polarization.

scholarly journals Extracellular nicotinamide phosphoribosyltransferase (NAMPT) promotes M2 macrophage polarization in chronic lymphocytic leukemia

Blood ◽  
2015 ◽  
Vol 125 (1) ◽  
pp. 111-123 ◽  
Author(s):  
Valentina Audrito ◽  
Sara Serra ◽  
Davide Brusa ◽  
Francesca Mazzola ◽  
Francesca Arruga ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Chronic Lymphocytic Leukemia ◽  
Macrophage Polarization ◽  
Lymphocytic Leukemia ◽  
T Cell Proliferation ◽  
M2 Macrophage ◽  
M2 Macrophages ◽  
Nicotinamide Phosphoribosyltransferase ◽  
Key Points ◽  
M2 Macrophage Polarization

Key Points CLL lymphocytes show high intracellular and extracellular NAMPT levels, further increased upon activation. eNAMPT prompts differentiation of CLL monocytes into M2 macrophages that sustain CLL survival and reduce T-cell proliferation.

scholarly journals FPR2 participates in epithelial ovarian cancer (EOC) progression through RhoA-mediated M2 macrophage polarization

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Xiaohui Xie ◽  
Juan He ◽  
Qiong Wang ◽  
Yaqiong Liu ◽  
Weiwei Chen ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Western Blot ◽  
Epithelial Ovarian Cancer ◽  
Cell Line ◽  
Cell Lines ◽  
Macrophage Polarization ◽  
M2 Macrophage ◽  
M2 Macrophages ◽  
Invasion And Metastasis ◽  
M2 Macrophage Polarization

Abstract Background In our previous study, we found that formyl peptide receptor 2 (FPR2) promoted the invasion and metastasis of epithelial ovarian cancer (EOC) and could be a prognostic marker for EOC. In this study, we aimed to study the possible mechanism of FPR2 in promoting EOC progression. Methods EOC cell lines with ectopic FPR2 expression and knockdown as well as their control cell lines were established, and the expression change of RhoA in each cell line was evaluated by real time quantitative polymerase chain reaction (RT-qPCR) and Western blot. Wound healing and Transwell assays were performed to detect the migratory ability of EOCs affected by FPR2 and RhoA. The supernatant of each EOC cell line was used to coculture with macrophages, and then we tested M1 and M2 macrophage biomarkers in the supernatants by flow cytometry. The THP-1 cell line was also induced to differentiate into M1 and M2 macrophages, and FPR2 and RhoA expression in each macrophage cell line was detected by RT-qPCR and Western blot. A tumour xenograft model was established with SKOV3 and SKOV3−shFPR2 cell lines, and tumour volumes and weights were recorded. Results RhoA expression was significantly increased in EOCs along with the overexpression of FPR2, which showed a positive correlation by Pearson correlation analysis. Ectopic FPR2 expression contributes to the migratory ability of EOCs, and a RhoA inhibitor (C3 transferase) impairs EOC migration. Furthermore, FPR2 stimulated the secretion of Th2 cytokines by EOCs, which induced macrophages to differentiate to the M2 phenotype, while a RhoA inhibitor stimulated the secretion of Th1 cytokines and induced macrophages to differentiate to the M1 phenotype. Moreover, compared with M1 macrophages and THP-1 cells, FPR2 and RhoA expression was significantly upregulated in M2 macrophages. Conclusion FPR2 stimulated M2 macrophage polarization and promoted invasion and metastasis of ovarian cancer cells through RhoA.

scholarly journals Hexapeptide induces M2 macrophage polarization via the JAK1/STAT6 pathway to promote angiogenesis in bone repair

2021 ◽  
Author(s):  
Xinyun Han ◽  
Junxian Hu ◽  
Wenbo Zhao ◽  
Hongwei Lu ◽  
Jingjin Dai ◽  
...  
Keyword(s):  
Bone Repair ◽  
Macrophage Polarization ◽  
M2 Macrophage ◽  
M2 Macrophages ◽  
Promising Alternative ◽  
Defect Repair ◽  
M2 Macrophage Polarization ◽  
Underlying Mechanisms ◽  
Main Regulator

Angiogenesis is essential for successful bone defect repair. In normal tissue repair, the physiological inflammatory response is the main regulator of angiogenesis through the activity of macrophages and the cytokines secreted by them. In particular, M2 macrophages which secrete high levels of PDGF-BB are typically considered to promote angiogenesis. A hexapeptide [WKYMVm, (Trp-Lys-Tyr-Met-Val-D-Met-NH2)] has been reported to modulate inflammatory activities. However, the underlying mechanisms by which WKYMVm regulates macrophages remain unclear. In this study, the possible involvement by which WKYMVm induces the polarization of macrophages and affects their behaviors was evaluated. In vitro results showed that macrophages were induced to an M2 rather than M1 phenotype and the M2 phenotype was enhanced by WKYMVm through activation of the JAK1/STAT6 signaling pathway. It was also found that WKYMVm played an important role in the PDGF-BB production increase and proangiogenic abilities in M2 macrophages. Consistent with the results in vitro, the elevated M2/M0 ratio induced by WKYMVm enhanced the formation of new blood vessels in a femoral defect mouse model. In summary, these findings suggest that WKYMVm could be a promising alternative strategy for angiogenesis in bone repair by inducing M2 macrophage polarization.

scholarly journals Magnoliae Cortex Alleviates Muscle Wasting by Modulating M2 Macrophages in a Cisplatin-Induced Sarcopenia Mouse Model

2021 ◽  
Vol 22 (6) ◽  
pp. 3188
Author(s):  
Minwoo Hong ◽  
Ik-Hwan Han ◽  
Ilseob Choi ◽  
Nari Cha ◽  
Woojin Kim ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Macrophage Polarization ◽  
M2 Macrophage ◽  
Rapid Weight Loss ◽  
M2 Macrophages ◽  
Muscle Weight ◽  
Alternatively Activated Macrophages ◽  
Tnf Α ◽  
M2 Macrophage Polarization ◽  
Anti Inflammatory

Cachexia causes high mortality, low quality of life, and rapid weight loss in cancer patients. Sarcopenia, a condition characterized by the loss of muscle, is generally present in cachexia and is associated with inflammation. M2 macrophages, also known as an anti-inflammatory or alternatively activated macrophages, have been shown to play a role in muscle repair. Magnoliae Cortex (M.C) is a widely used medicinal herb in East Asia reported to have a broad range of anti-inflammatory activities; however, the effects of M.C on sarcopenia and on M2 macrophage polarization have to date not been studied. This study was designed to investigate whether the oral administration of M.C could decrease cisplatin-induced sarcopenia by modulating M2 macrophage polarization in mice. C57BL/6 mice were injected intraperitoneally with cisplatin (2.5 mg/kg) to mimic chemotherapy-induced sarcopenia. M.C extract (50, 100, and 200 mg/kg) was administered orally every 3 days (for a total of 12 times). M.C (100 and 200 mg/kg) significantly alleviated the cisplatin-induced loss of body mass, skeletal muscle weight, and grip strength. In addition, M.C increased the expression of M2 macrophage markers, such as MRC1, CD163, TGF-β, and Arg-1, and decreased the expression of M1-specific markers, including NOS2 and TNF-α, in skeletal muscle. Furthermore, the levels of like growth factor-1(IGF-1), as well as the number of M2a and M2c macrophages, significantly increased in skeletal muscle after M.C administration. M.C did not interfere with the anticancer effect of cisplatin in colon cancer. Our results demonstrated that M.C can alleviate cisplatin-induced sarcopenia by increasing the number of M2 macrophages. Therefore, our findings suggest that M.C could be used as an effective therapeutic agent to reverse or prevent cisplatin-induced sarcopenia.

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