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1.
Cytokine ; 182: 156729, 2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-39126768

RESUMO

BACKGROUND: Numerous studies have shown that various cytokines are important factors affecting bone mineral density (BMD), but the causality between the two remains uncertain. METHODS: Genetic variants associated with 41 circulating cytokines from a genome-wide association study (GWAS) in 8,293 Finns were used as instrumental variables (IVs) for a two-sample Mendelian randomization (MR) analysis. Inverse variance weighting (IVW) was employed as the primary method to investigate whether the 41 cytokines were causally associated with BMD at five different sites [total body bone mineral density (TB-BMD), heel bone mineral density (HE-BMD), forearm bone mineral density (FA-BMD), femoral neck bone mineral density (FN-BMD), and lumbar spine bone mineral density (LS-BMD)]. Weighted median and MR-Egger were chosen to further confirm the robustness of the results. We performed MR pleiotropy residual sum and outlier test (MR-PRESSO), MR-Egger regression, and Cochran's Q test to detect pleiotropy and sensitivity testing. RESULTS: After Bonferroni correction, two circulating cytokines had a strong causality with BMD at corresponding sites. Genetically predicted circulating hepatocyte growth factor (HGF) levels and HE-BMD were negatively correlated [ß (95 % CI) -0.035(-0.055, -0.016), P=0.00038]. Circulating macrophage inflammatory protein-1α (MIP-1α) levels and TB-BMD were negatively correlated [ß(95 %CI): -0.058(-0.092, -0.024), P=0.00074]. Weighted median and MR-Egger results were in line with the IVW results. We also found suggestive causal relationship (IVW P<0.05) between seven circulating cytokines and BMD at corresponding sites. No significant pleiotropy or heterogeneity was observed in our study. CONCLUSION: Our MR analyses indicated a causal effect between two circulating cytokines and BMD at corresponding sites (HGF and HE-BMD, MIP-1α and TB-BMD), along with suggestive evidence of a potential causality between seven cytokines and BMD at the corresponding sites. These findings would provide insights into the prevention and treatment of osteoporosis, especially immunoporosis.


Assuntos
Densidade Óssea , Citocinas , Estudo de Associação Genômica Ampla , Fator de Crescimento de Hepatócito , Análise da Randomização Mendeliana , Humanos , Densidade Óssea/genética , Citocinas/sangue , Masculino , Feminino , Fator de Crescimento de Hepatócito/sangue , Fator de Crescimento de Hepatócito/genética , Polimorfismo de Nucleotídeo Único/genética , Pessoa de Meia-Idade , Colo do Fêmur/metabolismo , Quimiocina CCL3/sangue , Quimiocina CCL3/genética
2.
Cell Transplant ; 33: 9636897241264912, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39076075

RESUMO

Wound healing is a complex process, which involves three stages: inflammation, proliferation, and remodeling. Inflammation is the first step; thus, immune factors play an important regulatory role in wound healing. In this study, we focused on a chemokine, C-C motif chemokine ligand 3 (CCL3), which is often upregulated for expression during wound healing. We compared cutaneous wound healing at the histological, morphological, and molecular levels in the presence and absence of CCL3. The results showed that the wound healing rate in the wild-type and CCL3-/- + CCL3 mice was faster than that of CCL3-/- mice (P < 0.01), and application of CCL3 to wounds increased the healing rate. In the process of wound healing, the degree of reepithelialization and the rate of collagen deposition in the wound of CCL3-/- mice were significantly lower than those of wild-type mice (P < 0.01). The number of macrophages and the expression levels of tumor necrosis factor(TNF)-α and transforming growth factor (TGF)-ß1 in the wounds of wild-type mice were much higher than those of the CCL3-/- mice. Removal of macrophages and CCL3-/- mice share similar phenotypes. Therefore, we infer that the wound healing requires the participation of macrophages, and CCL3 may play an important regulatory role through recruiting macrophages to the wound sites.


Assuntos
Quimiocina CCL3 , Macrófagos , Cicatrização , Animais , Quimiocina CCL3/metabolismo , Quimiocina CCL3/genética , Cicatrização/fisiologia , Macrófagos/metabolismo , Camundongos , Pele/patologia , Pele/metabolismo , Pele/lesões , Camundongos Endogâmicos C57BL , Fator de Crescimento Transformador beta1/metabolismo , Camundongos Knockout , Fator de Necrose Tumoral alfa/metabolismo , Masculino
3.
Connect Tissue Res ; 65(4): 330-342, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-39067006

RESUMO

BACKGROUND: Osteoarthritis (OA) is a degenerative disease that affects synovial joints and leads to significant pain and disability, particularly in older adults. Infiltration of macrophages plays a key role in the progression of OA. However, the mechanisms underlying macrophage recruitment in OA are not fully understood. METHODS: The Serglycin (SRGN) expression pattern was analyzed, along with its association with macrophage infiltration in OA, using bioinformatic methods. SRGN expression in chondrocytes was altered by small interfering RNA (siRNA) and plasmids. Conditioned media (CM) was obtained from transfected chondrocytes to establish a co-culture model of chondrocytes and THP-1 derived macrophages. The impact of SRGN on macrophage recruitment was evaluated using a transwell assay. Furthermore, the regulatory effect of SRGN on CCL3 was validated through qPCR, WB, and ELISA experiments. RESULTS: In OA patients, the upregulation of SRGN positively correlated with K-L grade and macrophage infiltration. It was found that SRGN expression and secretion were up-regulated in OA and that it can promote macrophage migration in vitro. Further investigation showed that SRGN affects macrophage migration by regulating the expression of CCL3. CONCLUSION: SRGN in chondrocytes plays a role in promoting the recruitment of THP-1 derived macrophages in vitro by regulating production of CCL3.


Assuntos
Quimiocina CCL3 , Condrócitos , Macrófagos , Osteoartrite , Proteínas de Transporte Vesicular , Humanos , Osteoartrite/patologia , Osteoartrite/metabolismo , Osteoartrite/genética , Quimiocina CCL3/metabolismo , Quimiocina CCL3/genética , Macrófagos/metabolismo , Macrófagos/patologia , Condrócitos/metabolismo , Condrócitos/patologia , Masculino , Proteínas de Transporte Vesicular/metabolismo , Proteínas de Transporte Vesicular/genética , Proteoglicanas/metabolismo , Feminino , Pessoa de Meia-Idade , Células THP-1 , Idoso , Movimento Celular
4.
PLoS One ; 19(7): e0304590, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39052611

RESUMO

BACKGROUND: Lipopolysaccharides (LPS) is well known to manifest a miscarriage-inducing effector during early pregnancy and activate macrophage to induce M1 macrophage polarization. However, the role of macrophage polarization in LPS-related miscarriage-inducing effect is not apparent. METHODS: In this work, gene expression changes and the percentage of M1/M2 macrophages and monocytes in LPS-induced miscarried uterus were firstly analyzed by RNA sequencing (RNA-seq) and Flow Cytometry. To explore the origin that contributes to M1/M2 macrophage differentiation, the expression of monocyte chemotactic protein (MCP-1), CCL3, and CCL4, chemokines related to monocyte/macrophage migration, was tested by quantitative real time PCR (qRT-PCR). RESULTS: We found that percentage of M1 macrophages rose, while the percentage of M2 macrophages declined down in the injected mice uterus. Meanwhile, the percentage of M1 and M2 macrophages showed no significant difference in the spleens of LPS injected mice compared to PBS injected control mice. Expression of Mcp-1, Ccl3, and Ccl4 and numbers of monocytes were remarkably up-regulated in LPS-induced miscarried mice uterus. CONCLUSION: These results indicated that polarization and proportion changes of macrophage in the uterus may contribute to miscarriage. Our work provides new evidence correlating the aberrant regulation of M1/M2 macrophage polarization with deleterious miscarriage-inducing effects. This will help us understand the roles of critical immune cell differentiation in maintaining normal pregnancy.


Assuntos
Aborto Espontâneo , Lipopolissacarídeos , Macrófagos , Útero , Feminino , Animais , Macrófagos/metabolismo , Macrófagos/imunologia , Lipopolissacarídeos/farmacologia , Camundongos , Útero/imunologia , Útero/metabolismo , Gravidez , Aborto Espontâneo/imunologia , Quimiocina CCL2/metabolismo , Quimiocina CCL2/genética , Diferenciação Celular , Monócitos/metabolismo , Monócitos/imunologia , Quimiocina CCL3/metabolismo , Quimiocina CCL3/genética , Polaridade Celular , Quimiocina CCL4/metabolismo , Quimiocina CCL4/genética
5.
Front Immunol ; 15: 1362012, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38698846

RESUMO

Objectives: Knee osteoarthritis (KOA) and certain inflammatory cytokines (such as interleukin 1 [IL-1] and tumor necrosis factor alpha [TNF-a]) are related; however, the causal relationship remains unclear. Here, we aimed to assess the causal relationship between 41 inflammatory cytokines and KOA using Mendelian randomization (MR). Methods: Two-sample bidirectional MR was performed using genetic variation data for 41 inflammatory cytokines that were obtained from European Genome-Wide Association Study (GWAS) data (n=8293). KOA-related genetic association data were also obtained from European GWAS data (n=40,3124). Inverse variance weighting (IVW), MR, heterogeneity, sensitivity, and multiple validation analyses were performed. Results: Granulocyte colony-stimulating factor (G-CSF) or colony-stimulating factor 3 (CSF-3) levels were negatively associated with the risk of developing KOA (OR: 0.93, 95%CI:0.89-0.99, P=0.015). Additionally, macrophage inflammatory protein-1 alpha (MIP-1A/CCL3) was a consequence of KOA (OR: 0.72, 95%CI:0.54-0.97, P=0.032). No causal relationship was evident between other inflammatory cytokines and KOA development. Conclusion: This study suggests that certain inflammatory cytokines may be associated with KOA etiology. G-CSF exerts an upstream influence on KOA development, whereas MIP-1A (CCL-3) acts as a downstream factor.


Assuntos
Citocinas , Estudo de Associação Genômica Ampla , Análise da Randomização Mendeliana , Osteoartrite do Joelho , Polimorfismo de Nucleotídeo Único , Humanos , Quimiocina CCL3/genética , Quimiocina CCL3/sangue , Citocinas/genética , Citocinas/sangue , Predisposição Genética para Doença , Mediadores da Inflamação/metabolismo , Osteoartrite do Joelho/genética
6.
Biochem Biophys Res Commun ; 722: 150156, 2024 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-38797155

RESUMO

Osteosarcoma, considered as the primary cause of malignant bone tumors in children, necessitates novel therapeutic strategies to enhance overall survival rates. KAT7, a histone acetyltransferase, exerts pivotal functions in gene transcription and immune modulation. In light of this, our study identified a significant upregulation of KAT7 in the mRNA and protein levels in human osteosarcoma, boosting cell proliferation in vivo and in vitro. In addition, KAT7-mediated H3K14ac activation induced MMP14 transcription, leading to increased expression and facilitation of osteosarcoma cell metastasis. Subsequent bioinformatics analyses highlighted a correlation between KAT7 and adaptive immune responses, indicating CCL3 as a downstream target of KAT7. Mechanistically, STAT1 was found to transcriptionally upregulate CCL3 expression. Furthermore, overexpression of KAT7 suppressed CCL3 secretions, whereas knockdown of KAT7 enhanced its release. Overall, these findings underscore the oncogenic role of KAT7 in regulating immune responses for osteosarcoma treatment.


Assuntos
Neoplasias Ósseas , Quimiocina CCL3 , Regulação Neoplásica da Expressão Gênica , Histona Acetiltransferases , Osteossarcoma , Fator de Transcrição STAT1 , Transdução de Sinais , Animais , Humanos , Camundongos , Neoplasias Ósseas/genética , Neoplasias Ósseas/metabolismo , Neoplasias Ósseas/patologia , Linhagem Celular Tumoral , Proliferação de Células/genética , Quimiocina CCL3/metabolismo , Quimiocina CCL3/genética , Histona Acetiltransferases/metabolismo , Histona Acetiltransferases/genética , Camundongos Nus , Osteossarcoma/genética , Osteossarcoma/metabolismo , Osteossarcoma/patologia , Fator de Transcrição STAT1/metabolismo , Fator de Transcrição STAT1/genética
7.
Fish Shellfish Immunol ; 149: 109614, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38710342

RESUMO

Chemokines are critical molecules involved in immune reaction and immune system homeostasis, and some chemokines play a role in antiviral immunity. It is not known if the C-C motif chemokine ligand 3 (CCL3), a member of the CC chemokine family, possesses antiviral properties in fish. In this study, a ccl3 was cloned from the mandarin fish (Siniperca chuatsi), and it has an open reading frame (ORF) of 276 base pairs, which are predicted to encode a 91-amino acid peptide. Mandarin fish CCL3 revealed conserved sequence features with four cysteine residues and closely relationships with the CCL3s from other vertebrates based on the sequence alignment and phylogenetic analysis. The transcripts of ccl3 were notably enriched in immune-related organs, such as spleen and gills in healthy mandarin fish, and the ccl3 was induced in the isolated mandarin fish brain (MFB) cells following infection with infectious spleen and kidney necrosis virus (ISKNV). Moreover, in MFB cells, overexpression of CCL3 induced immune factors, such as IL1ß, TNFα, MX, IRF1 and IFNh, and exhibited antiviral activity against ISKNV. This study sheds light on the immune role of CCL3 in immune response of mandarin fish, and its antiviral defense mechanism is of interest for further investigation.


Assuntos
Sequência de Aminoácidos , Infecções por Vírus de DNA , Doenças dos Peixes , Proteínas de Peixes , Imunidade Inata , Iridoviridae , Perciformes , Filogenia , Alinhamento de Sequência , Animais , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Proteínas de Peixes/química , Doenças dos Peixes/imunologia , Doenças dos Peixes/virologia , Perciformes/imunologia , Perciformes/genética , Infecções por Vírus de DNA/imunologia , Infecções por Vírus de DNA/veterinária , Iridoviridae/fisiologia , Alinhamento de Sequência/veterinária , Imunidade Inata/genética , Regulação da Expressão Gênica/imunologia , Quimiocina CCL3/genética , Quimiocina CCL3/imunologia , Clonagem Molecular , Perfilação da Expressão Gênica/veterinária , Sequência de Bases
8.
Cell Commun Signal ; 22(1): 263, 2024 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-38730482

RESUMO

BACKGROUND: Helicobacter pylori (H. pylori) is the predominant etiological agent of gastritis and disrupts the integrity of the gastric mucosal barrier through various pathogenic mechanisms. After H. pylori invades the gastric mucosa, it interacts with immune cells in the lamina propria. Macrophages are central players in the inflammatory response, and H. pylori stimulates them to secrete a variety of inflammatory factors, leading to the chronic damage of the gastric mucosa. Therefore, the study aims to explore the mechanism of gastric mucosal injury caused by inflammatory factors secreted by macrophages, which may provide a new mechanism for the development of H. pylori-related gastritis. METHODS: The expression and secretion of CCL3 from H. pylori infected macrophages were detected by RT-qPCR, Western blot and ELISA. The effect of H. pylori-infected macrophage culture medium and CCL3 on gastric epithelial cells tight junctions were analyzed by Western blot, immunofluorescence and transepithelial electrical resistance. EdU and apoptotic flow cytometry assays were used to detect cell proliferation and apoptosis levels. Dual-luciferase reporter assays and chromatin immunoprecipitation assays were used to study CCL3 transcription factors. Finally, gastric mucosal tissue inflammation and CCL3 expression were analyzed by hematoxylin and eosin staining and immunohistochemistry. RESULTS: After H. pylori infection, CCL3 expressed and secreted from macrophages were increased. H. pylori-infected macrophage culture medium and CCL3 disrupted gastric epithelial cells tight junctions, while CCL3 neutralizing antibody and receptor inhibitor of CCL3 improved the disruption of tight junctions between cells. In addition, H. pylori-infected macrophage culture medium and CCL3 recombinant proteins stimulated P38 phosphorylation, and P38 phosphorylation inhibitor improved the disruption of tight junctions between cells. Besides, it was identified that STAT1 was a transcription factor of CCL3 and H. pylori stimulated macrophage to secret CCL3 through the JAK1-STAT1 pathway. Finally, after mice were injected with murine CCL3 recombinant protein, the gastric mucosal injury and inflammation were aggravated, and the phosphorylation level of P38 was increased. CONCLUSIONS: In summary, our findings demonstrate that H. pylori infection stimulates macrophages to secrete CCL3 via the JAK1-STAT1 pathway. Subsequently, CCL3 damages gastric epithelial tight junctions through the phosphorylation of P38. This may be a novel mechanism of gastric mucosal injury in H. pylori-associated gastritis.


Assuntos
Quimiocina CCL3 , Mucosa Gástrica , Infecções por Helicobacter , Helicobacter pylori , Macrófagos , Helicobacter pylori/fisiologia , Quimiocina CCL3/metabolismo , Quimiocina CCL3/genética , Animais , Mucosa Gástrica/metabolismo , Mucosa Gástrica/patologia , Mucosa Gástrica/microbiologia , Macrófagos/metabolismo , Macrófagos/microbiologia , Camundongos , Infecções por Helicobacter/metabolismo , Infecções por Helicobacter/patologia , Homeostase , Camundongos Endogâmicos C57BL , Humanos , Apoptose , Proliferação de Células , Masculino , Células RAW 264.7
9.
Bone Res ; 12(1): 34, 2024 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-38816384

RESUMO

Degenerated endplate appears with cheese-like morphology and sensory innervation, contributing to low back pain and subsequently inducing intervertebral disc degeneration in the aged population.1 However, the origin and development mechanism of the cheese-like morphology remain unclear. Here in this study, we report lumbar instability induced cartilage endplate remodeling is responsible for this pathological change. Transcriptome sequencing of the endplate chondrocytes under abnormal stress revealed that the Hippo signaling was key for this process. Activation of Hippo signaling or knockout of the key gene Yap1 in the cartilage endplate severed the cheese-like morphological change and disc degeneration after lumbar spine instability (LSI) surgery, while blocking the Hippo signaling reversed this process. Meanwhile, transcriptome sequencing data also showed osteoclast differentiation related gene set expression was up regulated in the endplate chondrocytes under abnormal mechanical stress, which was activated after the Hippo signaling. Among the discovered osteoclast differentiation gene set, CCL3 was found to be largely released from the chondrocytes under abnormal stress, which functioned to recruit and promote osteoclasts formation for cartilage endplate remodeling. Over-expression of Yap1 inhibited CCL3 transcription by blocking its promoter, which then reversed the endplate from remodeling to the cheese-like morphology. Finally, LSI-induced cartilage endplate remodeling was successfully rescued by local injection of an AAV5 wrapped Yap1 over-expression plasmid at the site. These findings suggest that the Hippo signaling induced osteoclast gene set activation in the cartilage endplate is a potential new target for the management of instability induced low back pain and lumbar degeneration.


Assuntos
Quimiocina CCL3 , Via de Sinalização Hippo , Degeneração do Disco Intervertebral , Vértebras Lombares , Osteoclastos , Transdução de Sinais , Degeneração do Disco Intervertebral/patologia , Degeneração do Disco Intervertebral/metabolismo , Degeneração do Disco Intervertebral/genética , Animais , Osteoclastos/metabolismo , Osteoclastos/patologia , Vértebras Lombares/patologia , Quimiocina CCL3/genética , Quimiocina CCL3/metabolismo , Camundongos , Cartilagem/patologia , Cartilagem/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Serina-Treonina Quinases/genética , Instabilidade Articular/patologia , Instabilidade Articular/genética , Condrócitos/metabolismo , Condrócitos/patologia , Proteínas de Sinalização YAP/metabolismo , Masculino , Camundongos Endogâmicos C57BL
10.
J Exp Med ; 221(7)2024 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-38661718

RESUMO

Chemokines guide immune cells during their response against pathogens and tumors. Various techniques exist to determine chemokine production, but none to identify cells that directly sense chemokines in vivo. We have generated CCL3-EASER (ErAse, SEnd, Receive) mice that simultaneously report for Ccl3 transcription and translation, allow identifying Ccl3-sensing cells, and permit inducible deletion of Ccl3-producing cells. We infected these mice with murine cytomegalovirus (mCMV), where Ccl3 and NK cells are critical defense mediators. We found that NK cells transcribed Ccl3 already in homeostasis, but Ccl3 translation required type I interferon signaling in infected organs during early infection. NK cells were both the principal Ccl3 producers and sensors of Ccl3, indicating auto/paracrine communication that amplified NK cell response, and this was essential for the early defense against mCMV. CCL3-EASER mice represent the prototype of a new class of dual fluorescence reporter mice for analyzing cellular communication via chemokines, which may be applied also to other chemokines and disease models.


Assuntos
Comunicação Celular , Quimiocina CCL3 , Modelos Animais , Biossíntese de Proteínas , Transcrição Gênica , Animais , Camundongos , Comunicação Celular/imunologia , Quimiocina CCL3/genética , Quimiocina CCL3/imunologia , Técnicas de Introdução de Genes , Camundongos Transgênicos , Muromegalovirus , Biossíntese de Proteínas/efeitos dos fármacos , Biossíntese de Proteínas/imunologia , Transcrição Gênica/imunologia , Células Matadoras Naturais/imunologia , Interferon beta/farmacologia , Infecções por Herpesviridae/imunologia
11.
Int J Mol Sci ; 25(8)2024 Apr 18.
Artigo em Inglês | MEDLINE | ID: mdl-38674048

RESUMO

Inflammation processes of the central nervous system (CNS) play a vital role in the pathogenesis of several neurological and psychiatric disorders like depression. These processes are characterized by the activation of glia cells, such as microglia. Clinical studies showed a decrease in symptoms associated with the mentioned diseases after the treatment with anti-inflammatory drugs. Therefore, the investigation of novel anti-inflammatory drugs could hold substantial potential in the treatment of disorders with a neuroinflammatory background. In this in vitro study, we report the anti-inflammatory effects of a novel hexacyclic peptide-peptoid hybrid in lipopolysaccharide (LPS)-stimulated BV2 microglial cells. The macrocyclic compound X15856 significantly suppressed Interleukin 6 (IL-6), tumor necrosis factor-α (TNF-α), c-c motif chemokine ligand 2 (CCL2), CCL3, C-X-C motif chemokine ligand 2 (CXCL2), and CXCL10 expression and release in LPS-treated BV2 microglial cells. The anti-inflammatory effects of the compound are partially explained by the modulation of the phosphorylation of p38 mitogen-activated protein kinases (MAPK), p42/44 MAPK (ERK 1/2), protein kinase C (PKC), and the nuclear factor (NF)-κB, respectively. Due to its remarkable anti-inflammatory properties, this compound emerges as an encouraging option for additional research and potential utilization in disorders influenced by inflammation, such as depression.


Assuntos
Anti-Inflamatórios , Lipopolissacarídeos , Microglia , Microglia/efeitos dos fármacos , Microglia/metabolismo , Animais , Camundongos , Anti-Inflamatórios/farmacologia , Linhagem Celular , Peptoides/farmacologia , Peptoides/química , Interleucina-6/metabolismo , NF-kappa B/metabolismo , Quimiocina CCL2/metabolismo , Quimiocina CCL2/genética , Peptídeos/farmacologia , Peptídeos/química , Fator de Necrose Tumoral alfa/metabolismo , Quimiocina CXCL2/metabolismo , Citocinas/metabolismo , Doenças Neuroinflamatórias/tratamento farmacológico , Doenças Neuroinflamatórias/metabolismo , Quimiocina CCL3/metabolismo , Quimiocina CCL3/genética , Compostos Macrocíclicos/farmacologia , Compostos Macrocíclicos/química
12.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 40(3): 215-221, 2024 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-38512031

RESUMO

Objective To compare the functional differences between bone marrow derived macrophages and peritoneal macrophages, which may provide the basis for the selection of macrophages in immunological research and immunoregulatory drug evaluation. Methods Marophage colony-stimulating factor (M-CSF) was used to induce the differentiation of bone marrow monocytes into macrophages, and thioglycolate medium was used to induce peritonitis to obtain peritoneal macrophages. After both macrophages being stimulated by zymosan, LPS, R848 and CpG respectively, mRNA levels of tumor necrosis factor α(TNF-α), interleukin 6(IL-6), macrophage inflammatory protein 1α(MIP-1α), monocyte chemoattractant protein 1(MCP-1) were measured by Real-time fluorescent quantitative PCR and the concentrations of secreted TNF-α, IL-6, MIP-1α and MCP-1 were detected by ELISA. In addition, the expression of costimulatory molecules CD80, CD86, CD40 and histocompatibility complex II (MHC II) on the cell surface was analyzed by flow cytometry. Results After inducing by different TLR ligands, mRNA expression levels of inflammatory cytokines and chemokines were increased in both macrophages. The secretion of TNF-α, IL-6, MIP-1α and MCP-1 in peritoneal macrophages and the expression of CD86 and MHC II on the surface of peritoneal macrophages were significantly higher than those of bone marrow derived macrophages. Conclusion There are significant differences in the expression of inflammatory factors, chemokines, costimulatory molecules, and histocompatibility complex between bone marrow derived macrophages and peritoneal macrophages. Peritoneal macrophages have more complete macrophage function and is more suitable for immunological research and immunomodulatory drug evaluation.


Assuntos
Medula Óssea , Cavidade Peritoneal , Animais , Camundongos , Quimiocina CCL3/genética , Interleucina-6 , Fator de Necrose Tumoral alfa , Macrófagos , Antígeno B7-1 , Antígenos CD40 , RNA Mensageiro
13.
Braz Dent J ; 34(3): 129-135, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37466520

RESUMO

This case-control study evaluated the gene expression levels of interleukin (IL)-4, macrophage inflammatory protein type 1 alpha (MIP-1α), and metalloproteinase (MMP)-9, factors involved in the formation of giant cells in healthy peri-implant tissue and peri-implantitis. Thirty-five subjects (15 healthy and 20 with peri-implantitis), who met the inclusion and exclusion criteria, were included in this study. The peri-implant tissue biopsies were subjected to total RNA extraction, DNAse treatment, and cDNA synthesis. Subsequently, the reaction of real-time PCR was performed to evaluate the gene expression levels of IL-4, MIP-1α, and MMP-9 concerning the reference gene. IL-4 gene expression showed higher (18-fold) values in the Peri-Implantitis Group of Patients when compared with the Healthy (Control) Group (p<0.0001). Although MIP- 1α and MMP-9 gene expression levels were higher in diseased implants, they showed no significant differences (p=0.06 and p=0.2337), respectively. Within the limitations of this study, the results showed that in tissues affected by peri-implantitis, only levels of Il-4 were increased when compared with tissues in the control group.


Assuntos
Implantes Dentários , Peri-Implantite , Humanos , Peri-Implantite/genética , Peri-Implantite/metabolismo , Peri-Implantite/patologia , Quimiocina CCL3/genética , Interleucina-4/genética , Estudos de Casos e Controles , Metaloproteinase 9 da Matriz/genética , Expressão Gênica
14.
Acta Haematol ; 146(4): 277-286, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37015191

RESUMO

INTRODUCTION: Both microenvironmental signals from surrounding cells and changes in the genome of leukemic cells play essential role in the development of chronic lymphocytic leukemia. Nurse-like cells (NLCs) are one of the important elements of the microenvironment of CLL cells. The key role in the interactions of leukemic cells with NLCs is played by chemokines, which may interfere with the programmed cell death process in the leukemic lymphocytes. The aim of our study was analysis of selected microenvironmental factors having a potential impact on the leukemic cells survival, as well as their association with clinical, cytogenetic, and molecular parameters. For this study, we selected three types of molecules which can modulate microenvironment: chemokines IL-8 and CCL3 (which are classically secreted to extracellular matrix), soluble forms of adhesion molecules JAG1 and CD163, and secreted form of endogenous protein BIRC5. We assessed their expression in the serum of CLL patients as well as in medium of long-term NLCs cultures. METHODS: Long-term cell culture was prepared from mononuclear cells derived from the blood of 34 patients with CLL. Number of NLCs cells was evaluated, under a light inverted microscope. The concentration of IL-8, CCL3, sBIRC5, sCD163, and sJAG1 in culture medium and serum was assessed by enzyme-linked immunosorbent assays. RESULTS: There were significant differences in the concentration of IL-8, sBIRC5, CCL3, sCD163, and sJAG1 between the patient's blood serum and the culture medium. The concentrations of IL-8, CCL3, and JAG1 were higher in the culture medium, which confirmed the role of the microenvironment in the production of these proteins. In addition, the concentration of CCL3 chemokine in both patient's blood serum and in the culture medium correlated with the number of NLCs and with known prognostic factors in the course of CLL, e.g., Rai stage, WBC, expression of ZAP-70, CD38, and CD5/19. CONCLUSION: The microenvironment of CLL cells, which includes NLCs, plays an important role in the pathogenesis of CLL. The CCL3 chemokine seems to be a good factor representing microenvironment of CLL cells. Chronic lymphocytic leukemia is a complex and very heterogeneous disease; therefore, its progress should be considered both in the context of genetic changes and the interaction with microenvironmental cells.


Assuntos
Leucemia Linfocítica Crônica de Células B , Humanos , Quimiocina CCL3/genética , Quimiocina CCL3/metabolismo , Ensaio de Imunoadsorção Enzimática , Interleucina-8 , Leucemia Linfocítica Crônica de Células B/diagnóstico , Leucemia Linfocítica Crônica de Células B/genética , Leucemia Linfocítica Crônica de Células B/metabolismo , Prognóstico , Microambiente Tumoral/genética
15.
Front Immunol ; 14: 1048790, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36993968

RESUMO

COVID-19 induces chromatin remodeling in host immune cells, and it had previously been shown that vitamin B12 downregulates some inflammatory genes via methyl-dependent epigenetic mechanisms. In this work, whole blood cultures from moderate or severe COVID-19 patients were used to assess the potential of B12 as adjuvant drug. The vitamin normalized the expression of a panel of inflammatory genes still dysregulated in the leukocytes despite glucocorticoid therapy during hospitalization. B12 also increased the flux of the sulfur amino acid pathway, that regulates the bioavailability of methyl. Accordingly, B12-induced downregulation of CCL3 strongly and negatively correlated with the hypermethylation of CpGs in its regulatory regions. Transcriptome analysis revealed that B12 attenuates the effects of COVID-19 on most inflammation-related pathways affected by the disease. As far as we are aware, this is the first study to demonstrate that pharmacological modulation of epigenetic markings in leukocytes favorably regulates central components of COVID-19 physiopathology.


Assuntos
COVID-19 , Metilação de DNA , Epigênese Genética , Mediadores da Inflamação , Leucócitos , Vitamina B 12 , Vitamina B 12/farmacologia , Vitamina B 12/uso terapêutico , COVID-19/genética , COVID-19/imunologia , Metilação de DNA/efeitos dos fármacos , Inflamação/tratamento farmacológico , Inflamação/genética , Inflamação/imunologia , Humanos , Masculino , Feminino , Pessoa de Meia-Idade , Idoso , Mediadores da Inflamação/metabolismo , Leucócitos/efeitos dos fármacos , Leucócitos/metabolismo , Quimiocina CCL3/genética , Transcriptoma , Regulação para Baixo
16.
Eur J Trauma Emerg Surg ; 49(2): 1035-1045, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36227355

RESUMO

BACKGROUND: The miRNA profile is changed after burn or sepsis and is involved in regulating inflammatory reactions. However, the function and molecular mechanism of miRNAs in regulating burn sepsis-induced acute kidney injury (AKI) are still unclear. METHODS: In this study, animal and cell sepsis models were established after burned rats were injected with lipopolysaccharide (LPS) or NRK-52E cells treated with LPS, respectively. Cytokine expression, inflammatory cell infiltration, serum creatinine (Scr) and kidney injury molecule-1 (KIM-1) levels were analysed after the indicated treatments. RESULTS: Burn sepsis increased the expression of inflammatory factors (TNF-α and IL-1ß) and chemokines (MIP-1α, MIP-2 and MCP-1). Moreover, burn sepsis promoted macrophage and neutrophil infiltration into the kidney and upregulated the levels of Scr and KIM-1 in the kidney and urine. Ectopic expression of miR-181c significantly reduced LPS-induced TLR4 protein expression, suppressed KIM-1 mRNA levels and subsequently inhibited the activation of inflammatory genes (TNF-α and IL-1ß) and chemokine genes (MIP-1α, MIP-2 and MCP-1). CONCLUSIONS: Our results demonstrated that miR-181c could suppress TLR4 expression, reduce inflammatory factor and chemokine secretion, mitigate inflammatory cell infiltration into the kidney and downregulate KIM-1 expression, which might ultimately attenuate burn sepsis-induced AKI.


Assuntos
Injúria Renal Aguda , Queimaduras , MicroRNAs , Sepse , Animais , Ratos , Injúria Renal Aguda/etiologia , Queimaduras/complicações , Quimiocina CCL3/genética , Quimiocinas , Lipopolissacarídeos , MicroRNAs/genética , MicroRNAs/metabolismo , Sepse/complicações , Sepse/genética , Sepse/metabolismo , Receptor 4 Toll-Like/genética , Fator de Necrose Tumoral alfa/metabolismo
17.
JCI Insight ; 8(1)2023 01 10.
Artigo em Inglês | MEDLINE | ID: mdl-36378535

RESUMO

The central physiological role of the bone marrow renders bone marrow stromal cells (BMSCs) particularly sensitive to aging. With bone aging, BMSCs acquire a differentiation potential bias in favor of adipogenesis over osteogenesis, and the underlying molecular mechanisms remain unclear. Herein, we investigated the factors underlying age-related changes in the bone marrow and their roles in BMSCs' differentiation. Antibody array revealed that CC chemokine ligand 3 (CCL3) accumulation occurred in the serum of naturally aged mice along with bone aging phenotypes, including bone loss, bone marrow adiposity, and imbalanced BMSC differentiation. In vivo Ccl3 deletion could rescue these phenotypes in aged mice. CCL3 improved the adipogenic differentiation potential of BMSCs, with a positive feedback loop between CCL3 and C/EBPα. CCL3 activated C/EBPα expression via STAT3, while C/EBPα activated CCL3 expression through direct promoter binding, facilitated by DNA hypomethylation. Moreover, CCL3 inhibited BMSCs' osteogenic differentiation potential by blocking ß-catenin activity mediated by ERK-activated Dickkopf-related protein 1 upregulation. Blocking CCL3 in vivo via neutralizing antibodies ameliorated trabecular bone loss and bone marrow adiposity in aged mice. This study provides insights regarding age-related bone loss and bone marrow adiposity pathogenesis and lays a foundation for the identification of new targets for senile osteoporosis treatment.


Assuntos
Osteogênese , Osteoporose , Camundongos , Animais , Osteogênese/fisiologia , Adiposidade , Medula Óssea/patologia , Ligantes , Diferenciação Celular , Osteoporose/metabolismo , Obesidade/complicações , Quimiocina CCL3/genética
18.
Epilepsy Behav ; 137(Pt A): 108962, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-36356419

RESUMO

Neuroinflammation plays a protective role in the brain; however, in neurological diseases such as epilepsy, overactivated neuroinflammation, along with overexpression of inflammatory mediators, can cause neuronal tissue damage, which can trigger seizures due to loss of ionic or neurotransmitter homeostasis. Therefore, we aimed to evaluate mRNA expression levels of proinflammatory cytokines, early growth response factor 3 (Egr3), and GABA A receptors in the hippocampus of naive audiogenic mutant tremor mice, and stimulated tremor mice after a seizure. Gene expression of Il-1ß, Il-6, Tnf-α, Ccl2, Ccl3, Egr3, Gabra1, and Gabra4 from hippocampal samples of naive and stimulated tremor mice were measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Relative to resistant mice, Ccl3 gene expression was increased and Il6 was decreased in the hippocampus of naïve tremor mice. Thirty minutes after a seizure, Ccl3 and Il-1ß mRNA expression were decreased (p < 0.0001; p = 0.0034, respectively) while Il6 was increased (p = 0.0052) in stimulated tremor mice, relative to naïve animals. In addition, Egr3, Gabra1, and Gabra4 mRNA expression was decreased in the hippocampus of naive tremor mice, relative to resistant mice, which increased 30 minutes after a seizure (p = 0.0496; p = 0.0447, and p = 0.0011, respectively), relative to naïve animals. In conclusion, overexpression of Ccl3 in the hippocampus of naive tremor mice, followed by downregulation soon after seizure in stimulated tremor mice, could be involved in changes in the blood-brain barrier (BBB) permeability in epilepsy. Il-1ß may be involved in hippocampal downregulation of GABA A receptors of naive tremor mice, characterizing an important mechanism in audiogenic seizures triggering. Hippocampal alterations of proinflammatory cytokines, Egr3, and GABA A receptors in tremor mice reinforce them as an alternative tool to modeling temporal lobe epilepsy.


Assuntos
Epilepsia Reflexa , Receptores de GABA-A , Camundongos , Animais , Receptores de GABA-A/metabolismo , Tremor/metabolismo , Convulsões/genética , Hipocampo/metabolismo , Epilepsia Reflexa/genética , RNA Mensageiro , Quimiocina CCL3/genética , Quimiocina CCL3/metabolismo
19.
Exp Eye Res ; 224: 109213, 2022 11.
Artigo em Inglês | MEDLINE | ID: mdl-36063964

RESUMO

Bacillus cereus (B. cereus) endophthalmitis is a vision-threatening bacterial infection. Uncontrolled inflammatory responses are the hallmark of this disease which cause irreversible damage to the retina. We recently reported C-X-C chemokines as a vital modulators which impacted the pathogenesis of this disease. Here, we investigated the impact of two highly upregulated C-C chemokines, CCL2 and CCL3, on intraocular inflammation this disease. B. cereus was injected into the eyes of C57BL/6J (WT), CCL2-/-, and CCL3-/- mice to induce endophthalmitis. Infected eyes were examined for bacterial growth, retinal function, and inflammation. Bacterial growth in CCL2-/- and CCL3-/- mice were similar, but retained retinal function was greater in CCL2-/- and CCL3-/- eyes compared to that of C57BL/6J eyes. The retinal architecture of infected eyes of CCL2-/- mice were conserved for a longer period of time than in infected CCL3-/- eyes. Infected CCL2-/- and CCL3-/- eyes had less inflammation than did infected C57BL/6J eyes. Based on these results, we assessed the efficacies of intravitreal anti-CCL2 or anti-CCL3 with or without the antibiotic gatifloxacin. Compared to infected untreated eyes, there was significantly less inflammation and greater retention of retinal function in eyes treated with anti-CCL2 or anti-CCL3 with gatifloxacin. This study showed that B. cereus endophthalmitis in CCL2-/- mice had a better clinical outcome than in CCL3-/- mice. Intravitreal administration of anti-CCL2 and anti-CCL3 with gatifloxacin significantly reduced inflammation and provided protection of retinal function. These results suggest that CCL2 and CCL3 are prospective anti-inflammatory targets that should be tested along with other antibiotics for treating Bacillus and perhaps other forms of endophthalmitis.


Assuntos
Bacillus , Quimiocina CCL2 , Endoftalmite , Infecções Oculares Bacterianas , Uveíte , Animais , Camundongos , Antibacterianos/uso terapêutico , Bacillus cereus , Quimiocina CCL3/genética , Eletrorretinografia , Endoftalmite/tratamento farmacológico , Endoftalmite/microbiologia , Infecções Oculares Bacterianas/tratamento farmacológico , Infecções Oculares Bacterianas/microbiologia , Gatifloxacina/uso terapêutico , Inflamação , Camundongos Endogâmicos C57BL , Quimiocina CCL2/genética
20.
Basic Clin Pharmacol Toxicol ; 131(5): 406-419, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36029292

RESUMO

Patients with ulcerative colitis (UC) experience diarrhoea, hematochezia and abdominal pain. UC is a well-known health challenge affecting 200-250 per 100 000 individuals worldwide, with a similar prevalence in both sexes and elevated upon activation of gut immune responses. We evaluated the potential therapeutic effects of cycloastragenol in experimentally induced UC rats and examined the modulation of sphingosine kinase (SphK), macrophage inflammatory protein (MIP)-1α and miR-143. We treated UC rats with 30 mg/kg cycloastragenol and assessed gene and protein expression levels of SphK, MIP-1α, B-cell lymphoma 2 (BCL2), BCL2-associated X (BAX), miR-143, NF-κB, tumour necrosis factor (TNF)-α and active caspase-3. Colon sections were examined using electron microscopy; additional sections were stained with haematoxylin-eosin or immunostained with anti-TNF-α and anti-caspase-3 antibodies. Electron microscopy of UC specimens revealed dark distorted goblet cell nuclei with disarranged mucus granules and a nondistinct brush border with atypical microvilli. Haematoxylin-eosin staining showed damaged intestinal glands, severe haemorrhage and inflammatory cell infiltration. Cycloastragenol treatment improved the induced morphological changes. In UC rats, cycloastragenol significantly reduced expression levels of SphK, MIP-1α, BAX, NF-κB, TNF-α and active caspase-3, associated with BCL2 and miR-143 overexpression. Therefore, cycloastragenol protects against UC by modulating SphK/MIP-1α/miR-143, subsequently deactivating inflammatory and apoptotic pathways.


Assuntos
Colite Ulcerativa , MicroRNAs , Animais , Quimiocina CCL3/genética , Colite Ulcerativa/induzido quimicamente , Colite Ulcerativa/tratamento farmacológico , Amarelo de Eosina-(YS)/uso terapêutico , Feminino , Masculino , MicroRNAs/genética , NF-kappa B/metabolismo , Fosfotransferases (Aceptor do Grupo Álcool) , Ratos , Sapogeninas , Inibidores do Fator de Necrose Tumoral , Fator de Necrose Tumoral alfa/metabolismo , Proteína X Associada a bcl-2/genética
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