RESUMO
PURPOSE: Tamoxifen, a widely used drug for breast cancer treatment, is associated with adverse effects on the liver, including the development of fatty liver. This study aimed to investigate the potential protective effect of caffeine against tamoxifen-induced fatty liver in Wistar rats. METHODS: Rats were divided into normal control, tamoxifen + saline, and tamoxifen + caffeine. Plasma samples were assessed for biochemical markers related to oxidative stress, inflammation, liver function, and cell damage. Additionally, liver histopathology was examined to quantify the extent of fatty infiltration. RESULTS: In the tamoxifen + saline group, elevated levels of plasma malondialdehyde (MDA), tumor necrosis factor-alpha (TNF-α), alanine aminotransferase (ALT), cytokeratin 18, and soluble ST2 were observed compared to the normal control group, indicating increased oxidative stress, inflammation, and liver injury (p < 0.01). Moreover, histopathological examination revealed a significant increase in fatty infiltration (p < 0.001). However, in the tamoxifen + caffeine group, these markers were markedly reduced (p < 0.05, p < 0.01), and fatty infiltration was significantly mitigated (p < 0.001). CONCLUSIONS: The findings suggest that caffeine administration attenuates tamoxifen-induced fatty liver in rats by ameliorating oxidative stress, inflammation, liver injury, and cell damage. Histopathological evidence further supports the protective role of caffeine. This study highlights the potential of caffeine as a therapeutic intervention to counter tamoxifen-induced hepatic complications, contributing to the optimization of breast cancer treatment strategies.
Assuntos
Cafeína , Fígado Gorduroso , Malondialdeído , Estresse Oxidativo , Ratos Wistar , Tamoxifeno , Animais , Cafeína/farmacologia , Cafeína/uso terapêutico , Tamoxifeno/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Malondialdeído/análise , Fígado Gorduroso/induzido quimicamente , Fígado Gorduroso/prevenção & controle , Fígado Gorduroso/tratamento farmacológico , Feminino , Fígado/efeitos dos fármacos , Fígado/patologia , Alanina Transaminase/sangue , Ratos , Antineoplásicos Hormonais/farmacologia , Fator de Necrose Tumoral alfa/sangue , Fator de Necrose Tumoral alfa/análise , Biomarcadores/sangue , Biomarcadores/análise , Modelos Animais de DoençasRESUMO
Osteosarcoma (OS) is the most frequent primary malignant bone tumour, whose heterogeneity represents a major challenge for common antitumour therapies. Inflammatory cytokines are known to be necessary for OS progression. Therefore, to optimise therapy, it is important to discover reliable biomarkers by identifying the mechanism generating OS and investigating the inflammatory pathways that support the undifferentiated state. In this work, we highlight the differences of epigenetic activities of IL-1ß and TNFα, and the susceptibility of TET-1 enzymatic inhibition, in tumour progression of three different OS cell lines. Investigating DNA methylation of IL-6 promoter and determining its expression, we found that TET enzymatic inhibition influences proliferation induced by inflammatory cytokines in OS cell lines. Moreover, Bobcat 339 treatment blocks IL-1ß epigenetic action on IL-6 promoter, while only partially those of TNFα as well as inhibits IL-1ß-dependent epithelial-mesenchymal transition (EMT) process, but only partially those of TNFα. In conclusion, this work highlights that IL-1ß and TNFα have different effects on DNA demethylation in OS cell lines, making DNA methylation a potential biomarker of disease. Specifically, in IL-1ß treatment, TET-1 inhibition completely blocks tumour progression, while in TNFα actions, it is only partially effective. Given that these two inflammatory pathways can be therapeutic targets for treating these tumours, knowledge of their distinct epigenetic behaviours can be useful for developing precise and specific therapeutic strategies for this disease.
Assuntos
Metilação de DNA , Epigênese Genética , Interleucina-1beta , Osteossarcoma , Proteínas Proto-Oncogênicas , Fator de Necrose Tumoral alfa , Humanos , Interleucina-1beta/genética , Interleucina-1beta/farmacologia , Epigênese Genética/efeitos dos fármacos , Epigênese Genética/genética , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/farmacologia , Metilação de DNA/genética , Metilação de DNA/efeitos dos fármacos , Linhagem Celular Tumoral , Proteínas Proto-Oncogênicas/genética , Osteossarcoma/genética , Osteossarcoma/tratamento farmacológico , Progressão da Doença , Regiões Promotoras Genéticas/efeitos dos fármacos , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Transição Epitelial-Mesenquimal/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Oxigenases de Função Mista/genética , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/genética , Interleucina-6/genética , Neoplasias Ósseas/genética , Neoplasias Ósseas/tratamento farmacológico , Neoplasias Ósseas/patologiaRESUMO
OBJECTIVE: Tumour necrosis factor (TNF)-α is a proinflammatory marker and has been shown to affect mitochondrial function in different tissues. We investigated the effect on adipose tissue (AT) inflammation and mitochondrial respiration in patients with hidradenitis suppurativa (HS) after 12 weeks of treatment with adalimumab, a TNF-α inhibitor. METHODS: We sampled blood and an AT biopsy from 13 patients with HS and 10 control subjects after an overnight fast. The patients were retested after at least 12 weeks of treatment with adalimumab (40 mg/week). We measured macrophage content and mitochondrial respiration in the AT and interleukin (IL)-1ß, IL-6, IL-10, high-sensitivity C-reactive protein (hsCRP), interferon-γ, TNF-α, adiponectin and leptin in plasma. Clinical scores and Dermatology Quality of Life Index (DLQI) were assessed. RESULTS: We found a higher anti-inflammatory macrophage content (CD206+) in the patient group compared with the control group, but no differences between before and after the intervention. No difference in mitochondrial respiration was observed. We observed higher plasma IL-6 and hsCRP concentrations in patients with HS compared to controls, with no differences before and after the intervention. The difference between controls and HS patients was abolished after the intervention. HS patients improved their DLQI after the intervention with no change in clinical scores. CONCLUSION: Treatment with adalimumab in patients with HS does not alter AT inflammation or mitochondrial respiratory capacity; however, we did see a higher content of anti-inflammatory macrophages in the patient group compared with the control group.
Assuntos
Adalimumab , Tecido Adiposo , Hidradenite Supurativa , Inflamação , Mitocôndrias , Humanos , Adalimumab/uso terapêutico , Adalimumab/farmacologia , Masculino , Hidradenite Supurativa/tratamento farmacológico , Hidradenite Supurativa/metabolismo , Feminino , Adulto , Mitocôndrias/metabolismo , Tecido Adiposo/metabolismo , Inflamação/tratamento farmacológico , Pessoa de Meia-Idade , Respiração Celular/efeitos dos fármacos , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Macrófagos/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Nephrotoxicity occurs when the body is exposed to certain drugs or toxins. When kidney damage occurs, the kidney fails to eliminate excess urine and waste. Solanesol (C45H74O) is a tri-sesquiterpenoid alcohol first isolated from tobacco, and it is widely distributed in plants of the Solanaceae family. Solanesol (SNL) is an intermediate in the synthesis of coenzyme Q10 (CoQ10), an antioxidant which protects nerve cells. This study investigated the protective effect of SNL at doses of 30 and 60 mg/kg in gentamicin-induced nephrotoxicity in Wistar albino rats. Animals were distributed into six groups and administered 100 mg/kg gentamicin-intraperitoneal injection for 14 days. Biochemical assessments were performed on kidney homogenate, blood, and serum. Treatment with SNL was shown as lower serum levels of creatinine, blood urea nitrogen (BUN), thiobarbituric acid reactive substances (TBARS), and Tumor necrosis factor alpha)TNF-α ((p < .001). It also restored reduced glutathione (GSH) and mitochondrial complex enzymatic activity as protective measures against gentamicin-induced nephrotoxicity. SNL were shown to reduce inflammation and oxidative stress markers (p < .001). Histological findings furtherly augmented the protective effects of SNL. Long-term SNL therapy also restored mitochondrial electron transport chain complex enzymes, such as complex-I (p < .001). In conclusion, these findings suggest that SNL can represent a protective therapeutic option for drug-induced nephrotoxicity, a long-term adverse effect of aminoglycoside antibiotics such as gentamicin.
Assuntos
Gentamicinas , Rim , Estresse Oxidativo , Ratos Wistar , Ubiquinona , Gentamicinas/toxicidade , Animais , Ubiquinona/análogos & derivados , Ubiquinona/farmacologia , Ubiquinona/uso terapêutico , Ratos , Estresse Oxidativo/efeitos dos fármacos , Masculino , Rim/efeitos dos fármacos , Rim/patologia , Rim/metabolismo , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Antioxidantes/farmacologia , Antioxidantes/uso terapêutico , Nefropatias/induzido quimicamente , Nefropatias/patologia , Nefropatias/prevenção & controle , Nefropatias/metabolismo , Glutationa/metabolismo , Creatinina/sangue , Fator de Necrose Tumoral alfa/metabolismo , Fator de Necrose Tumoral alfa/sangue , Nitrogênio da Ureia Sanguínea , Terpenos/farmacologia , Terpenos/uso terapêutico , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Antibacterianos/toxicidadeRESUMO
Introduction: Cytokine release syndrome (CRS) is one of the leading causes of mortality in patients with COVID-19 caused by the SARS-CoV-2 coronavirus. However, the mechanism of CRS induced by SARS-CoV-2 is vague. Methods: Using spike protein combined with IL-2, IFN-γ, and TNF-α to stimulate human peripheral blood mononuclear cells (PBMCs) to secrete CRS-related cytokines, the content of cytokines in the supernatant was detected, and the effects of NK, T, and monocytes were analyzed. Results: This study shows that dendritic cells loaded with spike protein of SARS-CoV-2 stimulate T cells to release much more interleukin-2 (IL-2,) which subsequently cooperates with spike protein to facilitate PBMCs to release IL-1ß, IL-6, and IL-8. These effects are achieved via IL-2 stimulation of NK cells to release tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ), as well as T cells to release IFN-γ Mechanistically, IFN-γ and TNF-α enhance the transcription of CD40, and the interaction of CD40 and its ligand stabilizes the membrane expression of toll-like receptor 4 (TLR4) that serves as a receptor of spike protein on the surface of monocytes. As a result, there is a constant interaction between spike protein and TLR4, leading to continuous activation of nuclear factor-κ-gene binding (NF-κB). Furthermore, TNF-α also activates NF-κB signaling in monocytes, which further cooperates with IFN-γ and spike protein to modulate NF-κB-dependent transcription of CRS-related inflammatory cytokines. Discussion: Targeting TNF-α/IFN-γ in combination with TLR4 may represent a promising therapeutic approach for alleviating CRS in individuals with COVID-19.
Assuntos
COVID-19 , Síndrome da Liberação de Citocina , Interleucina-2 , SARS-CoV-2 , Glicoproteína da Espícula de Coronavírus , Linfócitos T , Humanos , Glicoproteína da Espícula de Coronavírus/imunologia , Interleucina-2/metabolismo , Interleucina-2/imunologia , COVID-19/imunologia , SARS-CoV-2/imunologia , SARS-CoV-2/fisiologia , Síndrome da Liberação de Citocina/imunologia , Linfócitos T/imunologia , Linfócitos T/metabolismo , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Interferon gama/metabolismo , Interferon gama/imunologia , Receptor 4 Toll-Like/metabolismo , NF-kappa B/metabolismo , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Citocinas/metabolismo , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Increased MMP-9 expression in the tumor microenvironment (TME) plays a crucial role in the extracellular matrix remodeling to facilitate cancer invasion and metastasis. However, the mechanism of MMP-9 upregulation in TME remains elusive. Since TGF-ß and TNF-α levels are elevated in TME, we asked whether these two agents interacted to induce/augment MMP-9 expression. Using a well-established MDA-MB-231 breast cancer model, we found that the synergy between TGF-ß and TNF-α led to MMP-9 upregulation at the transcriptional and translational levels, compared to treatments with each agent alone. Our in vitro findings are corroborated by co-expression of elevated MMP-9 with TGF-ß and TNF-α in human breast cancer tissues. Mechanistically, we found that the MMP-9 upregulation driven by TGF-ß/TNF-α cooperativity was attenuated by selective inhibition of the TGF-ßRI/Smad3 pathway. Comparable outcomes were observed upon inhibition of TGF-ß-induced phosphorylation of Smad2/3 and p38. As expected, the cells defective in Smad2/3 or p38-mediated signaling did not exhibit this synergistic induction of MMP-9. Importantly, the inhibition of histone methylation but not acetylation dampened the synergistic MMP-9 expression. Histone modification profiling further identified the H3K36me2 as an epigenetic regulatory mark of this synergy. Moreover, TGF-ß/TNF-α co-stimulation led to increased levels of the transcriptionally permissive dimethylation mark at H3K36 in the MMP-9 promoter. Comparable outcomes were noted in cells deficient in NSD2 histone methyltransferase. In conclusion, our findings support a cooperativity model in which TGF-ß could amplify the TNF-α-mediated MMP-9 production via chromatin remodeling and facilitate breast cancer invasion and metastasis.
Assuntos
Neoplasias da Mama , Regulação Neoplásica da Expressão Gênica , Metaloproteinase 9 da Matriz , Metástase Neoplásica , Fator de Crescimento Transformador beta , Fator de Necrose Tumoral alfa , Humanos , Metaloproteinase 9 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/genética , Neoplasias da Mama/patologia , Neoplasias da Mama/metabolismo , Neoplasias da Mama/genética , Fator de Necrose Tumoral alfa/metabolismo , Feminino , Fator de Crescimento Transformador beta/metabolismo , Linhagem Celular Tumoral , Histonas/metabolismo , Metilação , Transdução de Sinais , Microambiente TumoralRESUMO
OBJECTIVES: Carcinoembryonic-antigen-related cell-adhesion molecule 1 (CEACAM1) is an adhesion molecule that acts as a coinhibitory receptor in the immune system. We previously demonstrated that CEACAM1 is predominantly expressed on peripheral blood neutrophils in patients with RA. The aim of the present study was to investigate the effects of Janus kinase inhibitors (JAKi) on cytokine-activated human neutrophils and CEACAM1 expression. METHODS: Peripheral blood neutrophils were obtained from healthy subjects. Isolated neutrophils were stimulated with tumor necrosis factor-alpha (TNF-α) or granulocyte-macrophage colony-stimulating factor (GM-CSF) in the presence or absence of JAKi. The expression of CEACAM1 in peripheral blood neutrophils was analyzed by flow cytometry. Protein phosphorylation of signal transducer and activator of transcription (STAT)1, STAT3, and STAT5 was assessed by western blot using phospho-specific antibodies. RESULTS: We found that TNF-α-induced CEACAM1 expression was marginally suppressed after pretreatment with pan-JAK inhibitor, tofacitinib. Moreover, TNF-α induced STAT1 and STAT3 phosphorylation at the late stimulation phase (4 to 16 h). The expressions of CEACAM1 on neutrophils were markedly up-regulated by GM-CSF not by interleukin (IL)-6 stimulation. All JAKi inhibited GM-CSF-induced CEACAM1 expressions on neutrophils, however, the inhibitory effects of baricitinib were larger compared to those of tofacitinib or filgotinib. Moreover, CEACAM1 was marginally upregulated in interferon (IFN)-γ stimulated neutrophils. Similarly, JAKi inhibited IFN-γ-induced CEACAM1 expressions on neutrophils. CONCLUSIONS: We demonstrated that JAKi prevent GM-CSF-induced CEACAM1 expression in neutrophils, and JAKi-induced inhibition depends on their selectivity against JAK isoforms. These findings suggest that JAKi can modulate the expression of CEACAM1 in cytokine-activated neutrophils, thereby limiting their activation.
Assuntos
Antígenos CD , Moléculas de Adesão Celular , Fator Estimulador de Colônias de Granulócitos e Macrófagos , Inibidores de Janus Quinases , Neutrófilos , Pirimidinas , Fator de Necrose Tumoral alfa , Humanos , Neutrófilos/metabolismo , Neutrófilos/imunologia , Neutrófilos/efeitos dos fármacos , Moléculas de Adesão Celular/metabolismo , Antígenos CD/metabolismo , Pirimidinas/farmacologia , Inibidores de Janus Quinases/farmacologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Fosforilação/efeitos dos fármacos , Piperidinas/farmacologia , Pirróis/farmacologia , Ativação de Neutrófilo/efeitos dos fármacos , Citocinas/metabolismo , Transdução de Sinais/efeitos dos fármacosRESUMO
BACKGROUND: Endothelial dysfunction is an integral pathophysiologic mechanism in sickle cell disease (SCD), and can lead to many complications. Sleep-disordered breathing (SDB) is a SCD complication with diverse incidence and pathophysiology. This study aimed to determine the prevalence of SDB in children with SCD and to assess its relation to endothelial dysfunction. METHODS: Sixty children with SCD and 60 healthy controls were enrolled. The levels of TNF-α, IL-6, and IL-17A were evaluated in the entire cohort using enzyme-linked immunosorbent assay (ELISA) kits. Polysomnography (PSG) was performed for all SCD patients after completion of the Pediatric Sleep Questionnaire (PSQ). RESULTS: TNF-α, IL-6, and IL-17A levels were significantly greater in children with SCD than in controls (p-values < 0.001, < 0.001, and 0.006, respectively). The PSQ revealed symptoms suggestive of SDB in 50 children with SCD (83.3%), and PSG revealed obstructive sleep apnea (OSA) in 44 children with SCD (73.3%); 22 patients had mild OSA, and 22 had moderate-to-severe OSA according to the apnea-hypopnea index (AHI). TNF-α was significantly greater in SCD children who reported heavy or loud breathing, trouble breathing or struggle to breathe, and difficulty waking up in the morning (p-values = 0.002, 0.002, and 0.031, respectively). The IL-6 levels were significantly greater in SCD children who stopped growing normally (p-value = 0.002). The levels of IL-6 and IL-17A were significantly greater in SCD children with morning headaches (p-values = 0.007 and 0.004, respectively). CONCLUSION: Children with SCD showed a high prevalence of SDB with significantly elevated levels of markers of endothelial function, highlighting the interplay of SDB and endothelial dysfunction in SCD.
Assuntos
Anemia Falciforme , Endotélio Vascular , Interleucina-6 , Polissonografia , Síndromes da Apneia do Sono , Fator de Necrose Tumoral alfa , Humanos , Anemia Falciforme/complicações , Anemia Falciforme/fisiopatologia , Masculino , Feminino , Criança , Síndromes da Apneia do Sono/fisiopatologia , Síndromes da Apneia do Sono/epidemiologia , Síndromes da Apneia do Sono/complicações , Egito/epidemiologia , Interleucina-6/sangue , Fator de Necrose Tumoral alfa/sangue , Estudos de Casos e Controles , Endotélio Vascular/fisiopatologia , Interleucina-17/sangue , Prevalência , Adolescente , Biomarcadores/sangue , Estudos TransversaisRESUMO
Phosphodiesterase 4 (PDE4) inhibitor is associated with a broad-spectrum anti-inflammatory mechanism. However, securing clinically efficacious doses with sufficient safety margins remains challenging due to class specific adverse events that are often unavoidable in the clinic. ART-648 is an orally available PDE4 inhibitor being developed for the treatment of inflammatory diseases. According to the estimated clinical doses based on an in vitro whole-blood assay, a phase I study was designed. The purpose of this phase I study was to assess the safety, tolerability, pharmacokinetics (PK), and pharmacodynamics (PD) following single and multiple administration of ART-648 in healthy subjects. PD was assessed by suppression of lipopolysaccharide-induced TNFα release in ex vivo whole-blood assay. In the single rising dose study, ART-648 was safe and well tolerated with a dose-proportional increase in exposures up to 4 mg. Single doses of ART-648 demonstrated dose-dependent PD response, indicating target engagement at 2-8 mg doses. In the multiple rising dose study, doses up to 4 mg BID after careful titration were well tolerated, while doses up to 6 mg BID were tolerated not in all but the majority of subjects. In conclusion, ART-648 exhibits a favorable PK profile with robust target engagement at clinically safe and tolerated doses identified in healthy subjects.
Assuntos
Relação Dose-Resposta a Droga , Voluntários Saudáveis , Inibidores da Fosfodiesterase 4 , Humanos , Inibidores da Fosfodiesterase 4/farmacocinética , Inibidores da Fosfodiesterase 4/administração & dosagem , Inibidores da Fosfodiesterase 4/efeitos adversos , Masculino , Adulto , Feminino , Pessoa de Meia-Idade , Adulto Jovem , Método Duplo-Cego , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Lipopolissacarídeos/administração & dosagem , Administração Oral , Sulfonamidas , para-AminobenzoatosRESUMO
Mycobacterium abscessus (Mab) is an opportunistic nontuberculous mycobacterium responsible of difficult-to-treat pulmonary infections in vulnerable patients, such as those suffering from Cystic Fibrosis (CF), where it represents a major cause of morbidity and mortality. Additionally, due to the intrinsic extensive antimicrobial resistance spectrum displayed by this species and the side effects reported for some available antibiotics, the therapeutic management of such infections remains extremely difficult. In the present study, we show that phosphatidylserine liposomes (PS-L) enhance intracellular mycobacterial killing of Mab infected human macrophages with functional or pharmacologically inhibited cystic fibrosis conductance regulator (CFTR), by a mechanism involving phagosome acidification and reactive oxygen species (ROS) production. Additionally, PS-L significantly reduce proinflammatory response of Mab infected macrophages in terms of NF-kB activation and TNF-α production, irrespective of CFTR inhibition. Altogether, these results represent the proof of concept for a possible future development of PS-L as a therapeutic strategy against difficult-to-treat Mab infection.
Assuntos
Lipossomos , Macrófagos , Infecções por Mycobacterium não Tuberculosas , Mycobacterium abscessus , Fagossomos , Fosfatidilserinas , Espécies Reativas de Oxigênio , Humanos , Mycobacterium abscessus/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Lipossomos/metabolismo , Macrófagos/microbiologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Macrófagos/imunologia , Fagossomos/microbiologia , Fagossomos/metabolismo , Fosfatidilserinas/metabolismo , Infecções por Mycobacterium não Tuberculosas/microbiologia , Fator de Necrose Tumoral alfa/metabolismo , Regulador de Condutância Transmembrana em Fibrose Cística/metabolismo , NF-kappa B/metabolismo , Fibrose Cística/microbiologiaRESUMO
BACKGROUND: The pathogenesis of osteoarthritis (OA) involves the progressive degradation of articular cartilage. Exosomes derived from mesenchymal stem cells (MSC-EXOs) have been shown to mitigate joint pathological injury by attenuating cartilage destruction. Optimization the yield and therapeutic efficacy of exosomes derived from MSCs is crucial for promoting their clinical translation. The preconditioning of MSCs enhances the therapeutic potential of engineered exosomes, offering promising prospects for application by enabling controlled and quantifiable external stimulation. This study aims to address these issues by employing pro-inflammatory preconditioning of MSCs to enhance exosome production and augment their therapeutic efficacy for OA. METHODS: The exosomes were isolated from the supernatant of infrapatellar fat pad (IPFP)-MSCs preconditioned with a pro-inflammatory factor, TNF-α, and their production was subsequently quantified. The exosome secretion-related pathways in IPFP-MSCs were evaluated through high-throughput transcriptome sequencing analysis, q-PCR and western blot analysis before and after TNF-α preconditioning. Furthermore, exosomes derived from TNF-α preconditioned IPFP-MSCs (IPFP-MSC-EXOsTNF-α) were administered intra-articularly in an OA mouse model, and subsequent evaluations were conducted to assess joint pathology and gait alterations. The expression of proteins involved in the maintenance of cartilage homeostasis within the exosomes was determined through proteomic analysis. RESULTS: The preconditioning with TNF-α significantly enhanced the exosome secretion of IPFP-MSCs compared to unpreconditioned MSCs. The potential mechanism involved the activation of the PI3K/AKT signaling pathway in IPFP-MSCs by TNF-α precondition, leading to an up-regulation of autophagy-related protein 16 like 1(ATG16L1) levels, which subsequently facilitated exosome secretion. The intra-articular administration of IPFP-MSC-EXOsTNF-α demonstrated superior efficacy in ameliorating pathological changes in the joints of OA mice. The preconditioning of TNF-α enhanced the up-regulation of low-density lipoprotein receptor-related protein 1 (LRP1) levels in IPFP-MSC-EXOsTNF-α, thereby exerting chondroprotective effects. CONCLUSION: TNF-α preconditioning constitutes an effective and promising method for optimizing the therapeutic effects of IPFP-MSCs derived exosomes in the treatment of OA.
Assuntos
Exossomos , Células-Tronco Mesenquimais , Osteoartrite , Fator de Necrose Tumoral alfa , Exossomos/metabolismo , Animais , Células-Tronco Mesenquimais/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Camundongos , Osteoartrite/terapia , Osteoartrite/metabolismo , Tecido Adiposo/citologia , Camundongos Endogâmicos C57BL , Masculino , Modelos Animais de Doenças , Cartilagem Articular/metabolismo , Transplante de Células-Tronco Mesenquimais/métodos , Células Cultivadas , HumanosRESUMO
Mesenchymal stromal/stem cells (MSC) have emerged as a promising therapeutic avenue for treating autoimmune diseases, eliciting considerable interest and discussion regarding their underlying mechanisms. This study revealed the distinctive ability of human umbilical cord MSC to aggregate within the lymph nodes of mice afflicted with autoimmune diseases, but this phenomenon was not observed in healthy mice. The specific distribution is driven by the heightened expression of the CCL21-CCR7 axis in mice with autoimmune diseases, facilitating the targeted homing of MSC to the lymph nodes. Within the lymph nodes, MSC exhibit a remarkable capacity to modulate Th17 cell function, exerting a pronounced anti-inflammatory effect. Transplanted MSC stimulates the secretion of L-amino-acid oxidase (LAAO), a response triggered by elevated levels of tumor necrosis factor-α (TNF-α) in mice with autoimmune diseases through the NF-κB pathway. The presence of LAAO is indispensable for the efficacy of MSC, as it significantly contributes to the inhibition of Th17 cells. Furthermore, LAAO-derived indole-3-pyruvic acid (I3P) serves as a potent suppressor of Th17 cells by activating the aryl hydrocarbon receptor (AHR) pathway. These findings advance our understanding of the global immunomodulatory effects exerted by MSC, providing valuable information for optimizing therapeutic outcomes.
Assuntos
L-Aminoácido Oxidase , Linfonodos , Células-Tronco Mesenquimais , Células Th17 , Animais , Células-Tronco Mesenquimais/metabolismo , Células Th17/imunologia , Células Th17/metabolismo , L-Aminoácido Oxidase/metabolismo , Linfonodos/metabolismo , Camundongos , Humanos , Camundongos Endogâmicos C57BL , Receptores CCR7/metabolismo , Receptores de Hidrocarboneto Arílico/metabolismo , Receptores de Hidrocarboneto Arílico/genética , NF-kappa B/metabolismo , Transplante de Células-Tronco Mesenquimais , Transdução de Sinais , Doenças Autoimunes/imunologia , Doenças Autoimunes/patologia , Fator de Necrose Tumoral alfa/metabolismo , Quimiocina CCL21/metabolismoRESUMO
Batroxobin, isolated from Bothrops moojeni, is a defibrinogenating agent used as a thrombin-like serine protease against fibrinogen for improving microcirculation. Here, we investigated whether, and if so, how batroxobin acts in concert with NK cells in terms of anti-tumor effects. CD3+/CD56+ NK cells were isolated and cultured from C57BL/6 mouse spleen. NK cells' viability was tested via Lactate dehydrogenase (LDH) assay. Lewis lung cancer cell (1*107 cell/ml) was used to build animal models. All animals were divided into five groups and treated with Batroxobin and NK cells respectively. HE staining was used to detect the pathological morphology of tumor tissue. The contents of fibrinogen and TNF-α in serum were determined by ELISA. The protein expression levels of MMP2, MMP9, VEGF and CD44 in tumor tissues were detected by Western Blot or immunohistochemistry. Compared with Control group, Tumor growth was not significantly affected in the group treated with Batroxobin or NK cells alone, However, tumor growth was significantly inhibited in the NK cell combined with the Batroxobin group. Serum levels of Fbg and TNF-αin mice treated with Batroxobin combined with NK cells dropped significantly, bringing them closer to normal levels. WB results showed that the expression levels of MMP2/9, VEGF and CD44 in Batroxobin combined with NK cell group also significantly decreased. Batroxobin combined with adoptive immunotherapy with NK cells significantly inhibited the growth of Lewis lung cancer in mice.
Assuntos
Batroxobina , Carcinoma Pulmonar de Lewis , Fibrinogênio , Imunoterapia Adotiva , Células Matadoras Naturais , Neoplasias Pulmonares , Camundongos Endogâmicos C57BL , Fator de Necrose Tumoral alfa , Animais , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/efeitos dos fármacos , Imunoterapia Adotiva/métodos , Neoplasias Pulmonares/imunologia , Neoplasias Pulmonares/terapia , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/tratamento farmacológico , Carcinoma Pulmonar de Lewis/imunologia , Carcinoma Pulmonar de Lewis/patologia , Carcinoma Pulmonar de Lewis/terapia , Batroxobina/farmacologia , Fibrinogênio/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Metaloproteinase 2 da Matriz/metabolismo , Camundongos , Fator A de Crescimento do Endotélio Vascular/metabolismo , Receptores de Hialuronatos/metabolismo , Linhagem Celular TumoralRESUMO
The pathogenesis of acute lung injury is complex. Studies have demonstrated the role of neutrophil extracellular traps (NETs) in the process of lipopolysaccharide (LPS)-induced acute lung injury (ALI). However, the underlying mechanism remains unclear. In this study, the regulation of Nrf2 in the formation of NETs, which was pathogenic in LPS-induced ALI, was identified by analyzing the levels of Cit-H3, lung function, lung tissue pathology, lung wet/dry ratio, the inflammatory cells, cytokines and proteins in the bronchoalveolar lavage fluid (BALF) and in addition, the activity of lung myeloperoxidase (MPO) was also measured. Results showed that the levels of Cit-H3 measured by western blot in Nrf2-knockout (KO) mice were higher compared with the WT mice after LPS stimulation. To further investigate the NETs formation was pathogenic during LPS-induced ALI, the Nrf2-KO mice were treated with DNase I. Results showed that DNase I improved lung function and lung tissue pathology and significantly reduced lung wet/dry ratio and proteins in the BALF. Besides, DNase I also attenuated the infiltration of inflammatory cells and the cytokines (TNF-α, IL-1ß) production in the BALF and the activity of lung MPO. Therefore, these results together indicate that Nrf2 may intervene in the release of NETs during LPS-induced ALI in mice.
Assuntos
Lesão Pulmonar Aguda , Líquido da Lavagem Broncoalveolar , Armadilhas Extracelulares , Lipopolissacarídeos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fator 2 Relacionado a NF-E2 , Animais , Lesão Pulmonar Aguda/metabolismo , Lesão Pulmonar Aguda/induzido quimicamente , Fator 2 Relacionado a NF-E2/metabolismo , Camundongos , Armadilhas Extracelulares/metabolismo , Líquido da Lavagem Broncoalveolar/química , Masculino , Peroxidase/metabolismo , Neutrófilos/metabolismo , Pulmão/metabolismo , Pulmão/patologia , Interleucina-1beta/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Desoxirribonuclease I/metabolismo , Citocinas/metabolismo , Western BlottingRESUMO
BACKGROUND: The Coronavirus Disease 2019 (COVID-19) pandemic has led to significant global morbidity and mortality. Understanding the genetic factors that influence disease outcomes can provide critical insights into pathogenesis and potential therapeutic targets. OBJECTIVE: This study aimed to investigate the potential correlation between single nucleotide polymorphisms (SNPs) in Interleukin 12 Subunit Alpha (IL-12A), Interleukin 12 Subunit Beta (IL-12B), Interleukin 6 (IL-6), and Tumor Necrosis Factor (TNF) genes and the severity as well as susceptibility to COVID-19 among Moroccan patients. PATIENTS AND METHODS: Next-Generation sequencing (NGS) was conducted on 325 Moroccan participants, 207 patients with PCR-confirmed Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection and 118 controls. Among these patients, 51% presented moderate to severe symptoms requiring hospitalization, while 49% were asymptomatic or experienced mild symptoms and did not require hospitalization. Statistical analysis was performed using codominant, dominant, and recessive logistic regression models to assess correlations with the severity and susceptibility to COVID-19 infection. RESULTS: No association was found between SNPs of IL-12A, IL-12B, IL-6 or TNF and COVID-19 severity and susceptibility. However, our results unveiled a noteworthy association with IL-6 rs2069840, which exhibited a negative correlation (OR = 0.21, 95% CI = 0.07-0.69, p = .006), suggesting a protective effect against SARS-CoV-2 infection. CONCLUSION: Polymorphisms in IL-12A, IL-12B, IL-6, and TNF genes are not correlated to the severity and susceptibility of COVID-19.
Assuntos
COVID-19 , Predisposição Genética para Doença , Subunidade p35 da Interleucina-12 , Subunidade p40 da Interleucina-12 , Interleucina-6 , Polimorfismo de Nucleotídeo Único , Índice de Gravidade de Doença , Fator de Necrose Tumoral alfa , Humanos , COVID-19/genética , COVID-19/imunologia , COVID-19/virologia , Interleucina-6/genética , Masculino , Feminino , Pessoa de Meia-Idade , Fator de Necrose Tumoral alfa/genética , Subunidade p40 da Interleucina-12/genética , Adulto , Subunidade p35 da Interleucina-12/genética , SARS-CoV-2 , Marrocos , Idoso , Estudos de Casos e ControlesRESUMO
Nanofluidic hybrid membranes display distinct ionic current rectification (ICR) properties and provide high surface area for immobilizing probes on the outer surface, exhibiting great potential in detection of biomolecules. Herein, we fabricated MOFs/AAO hybrid membrane with aptamers functionalized on the outer surface for in situ detection of living cells released secretions. TNF-α (a small molecular protein secreted by macrophages) was used as a model. After TNF-α was specifically captured by aptamers on the membrane surface, the asymmetry of surface charge on the hybrid membrane was amplified, the ICR was increased from 3.89 to 18.85. According to the ICR change, TNF-α was sensitively measured with a detection limit of â¼0.49 pM, which was significantly lower than other reported methods. When the hybrid membrane was clamped in the middle of self-made device, PET membrane incubated macrophages was rolled up and inserted into the chamber to mimic cellular microenvironment. Macrophages released TNF-α could be real time monitored with ionic current, macrophages and normal cells could be effectively distinguished according to the released TNF-α level. Thus, we proposed a nanofluidic platform for accurately measuring cell secretions in an engineered cellular microenvironment with a direct manner, without the need for labels or amplification steps.
Assuntos
Técnicas Biossensoriais , Transporte de Íons , Macrófagos , Fator de Necrose Tumoral alfa , Técnicas Biossensoriais/métodos , Técnicas Biossensoriais/instrumentação , Macrófagos/metabolismo , Macrófagos/citologia , Fator de Necrose Tumoral alfa/metabolismo , Fator de Necrose Tumoral alfa/análise , Estruturas Metalorgânicas/química , Animais , Aptâmeros de Nucleotídeos/química , Humanos , Desenho de Equipamento , Células RAW 264.7 , Camundongos , Membranas Artificiais , Limite de DetecçãoRESUMO
We developed a model of inflammation and airway remodeling in C57 mice provoked by exosomes derived from bone marrow mesenchymal stem cells infected by respiratory syncytial virus (RSV). The mean size of control and infected exosomes in vitro were 167.9 and 118.5 nm, respectively. After induction of modeled pathology, the severity of airway inflammation and its remodeling were analyzed by histopathological methods. In addition, the blood levels of inflammatory factors IL-10, IL-17, transforming growth factor-ß (TGF-ß), and TNFα were assayed; in the lung tissues, the expression levels of MMP-2, MMP-9, α-smooth muscle actin (α-SMA), and TGF-ß were measured. In the developed model, the effects of RSV-induced and non-induced exosomes were compared with those of inactivated and non-inactivated RSV. Intranasal administration of RSV-induced exosomes decreased the levels of serum inflammatory factors IL-10 and IL-17 and increased the expression of serum proinflammatory cytokine TNFα. Increased levels of MMP-2, MMP-9, and α-SMA, enhanced expression of TGF-ß in the lung tissue, and pathological staining of the lung tissues indicated infiltration with inflammatory cells and luminal constriction. Thus, RSV-induced exosomes can provoke airway inflammation and remodeling in mice similar to RSV, while non-induced exosomes cannot produce such alterations.
Assuntos
Remodelação das Vias Aéreas , Modelos Animais de Doenças , Exossomos , Interleucina-10 , Interleucina-17 , Metaloproteinase 2 da Matriz , Células-Tronco Mesenquimais , Camundongos Endogâmicos C57BL , Infecções por Vírus Respiratório Sincicial , Fator de Necrose Tumoral alfa , Animais , Exossomos/metabolismo , Células-Tronco Mesenquimais/metabolismo , Camundongos , Infecções por Vírus Respiratório Sincicial/patologia , Infecções por Vírus Respiratório Sincicial/imunologia , Infecções por Vírus Respiratório Sincicial/virologia , Infecções por Vírus Respiratório Sincicial/metabolismo , Metaloproteinase 2 da Matriz/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Fator de Necrose Tumoral alfa/sangue , Interleucina-10/metabolismo , Interleucina-10/sangue , Interleucina-17/metabolismo , Pulmão/patologia , Pulmão/virologia , Pulmão/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Vírus Sinciciais Respiratórios/patogenicidade , Fator de Crescimento Transformador beta/metabolismo , Actinas/metabolismo , Inflamação/patologia , Inflamação/metabolismo , Células da Medula Óssea/metabolismo , FemininoRESUMO
This study aimed to investigate the effects and potential mechanism of Polygonati Rhizoma aqueous extract on chronic obstructive pulmonary disease(COPD) in rats. Forty-eight Sprague-Dawley rats were randomly assigned to the normal, model,Yupingfeng Granules(1. 5 g·kg~(-1)), and low-, medium-, and high-dose(0. 25, 0. 5, and 1 g·kg~(-1), respectively) Polygonati Rhizoma aqueous extract groups. The rat model of COPD was established by cigarette smoke inhalation for 8 weeks, and then the modeled rats received corresponding treatment for 4 weeks. The grip strength and fecal moisture content were measured, and the lung index was calculated. Enzyme-linked immunosorbent assay(ELISA) was employed to determine the levels of interleukin(IL)-6 and tumor necrosis factor(TNF)-α in the lung tissue. Hematoxylin-eosin(HE) staining and Masson staining were performed to assess the pathological changes in the lung tissue. Flow cytometry was used to analyze T lymphocytes and their subpopulations in the peripheral blood, and the immunofluorescence assay and Western blot were employed to measure the protein levels of Toll-like receptor 4(TLR4), phosphorylated nuclear factor-kappaB(p-NF-κB), NF-κB, phosphorylated inhibitory kappa B-α(p-IκBα), IκBα, IL-6,and TNF-α in the lung tissue. The results indicated that the treatment with Polygonati Rhizoma aqueous extract significantly reduced the fecal moisture content, enhanced the grip strength, and inhibited inflammatory infiltration and fibrosis in the lung tissue. The treatment increased the Th/Tc ratio and Th cell proportion and decreased the Tc cell proportion in the peripheral blood. Furthermore,the treatment down-regulated the expression levels of TLR4, IL-6, and TNF-α and the p-NF-κB/NF-κB and p-IκBα/IκBα ratios in the lung tissue. In conclusion, Polygonati Rhizoma aqueous extract can ameliorate lung tissue damage in the rat model of COPD by inhibiting the TLR4/NF-κB signaling pathway and the production of inflammatory mediators.
Assuntos
Medicamentos de Ervas Chinesas , Pulmão , NF-kappa B , Polygonatum , Doença Pulmonar Obstrutiva Crônica , Ratos Sprague-Dawley , Rizoma , Receptor 4 Toll-Like , Animais , Ratos , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Medicamentos de Ervas Chinesas/farmacologia , Medicamentos de Ervas Chinesas/administração & dosagem , Medicamentos de Ervas Chinesas/química , Masculino , Polygonatum/química , NF-kappa B/metabolismo , NF-kappa B/genética , Receptor 4 Toll-Like/metabolismo , Receptor 4 Toll-Like/genética , Pulmão/efeitos dos fármacos , Rizoma/química , Interleucina-6/genética , Interleucina-6/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Fator de Necrose Tumoral alfa/genética , HumanosRESUMO
This study aims to investigate the mechanism of Xuanbai Chengqi Decoction in treating acute lung injury(ALI) based on network pharmacology and animal experiments. The potential targets and signaling pathways of Xuanbai Chengqi Decoction in regulating ALI were predicted by network pharmacology. The rat model of ALI was constructed and administrated with different doses of Xuanbai Chengqi Decoction. The pathological changes in the lung tissue of rats were observed by hematoxylin-eosin(HE) staining. The levels of interleukin-6(IL-6), interleukin-1ß(IL-1ß), and tumor necrosis factor-α(TNF-α) in the peripheral blood were measured by enzyme-linked immunosorbent assay(ELISA). The mRNA and protein levels of factors in the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR) signaling pathway were determined by quantitative real-time PCR(qPCR) and Western blot, respectively. A total of 52 compounds from Xuanbai Chengqi Decoction were predicted to be involved in the treatment of ALI, including ß-sitosterol, emodin, stigmasterol, glabridin, and aloe-emodin, which corresponded to 112 targets,and 4 723 targets of ALI were predicted. The compounds and ALI shared 94 common targets. The key targets included TNF, IL-1ß,prostaglandin-endoperoxide synthase 2(PTGS2), and tumor protein 53(TP53). Lipids and atherosclerosis, p53 signaling pathway,IL-17 signaling pathway, and PI3K/Akt signaling pathway were mainly involved in the treatment. Animal experiments showed that compared with the model group, Xuanbai Chengqi Decoction alleviated the pathological changes in the lung tissue, lowered the serum levels of IL-6, IL-1ß, and TNF-α, down-regulated the mRNA and protein levels of PI3K, Akt, and mTOR, and reduced the p-PI3K/PI3K, p-Akt/Akt, and p-mTOR/mTOR ratios in ALI rats. The results showed that Xuanbai Chengqi Decoction exerted its therapeutic effects on ALI via multiple components, targets, and pathways. Meanwhile, Xuanbai Chengqi Decoction may reduce the inflammation and attenuate the lung injuries of ALI rats by inhibiting the PI3K/Akt/mTOR signaling pathway.