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Because of the tumor's recurrence and significant metastasis, the standard single-therapy paradigm has failed to meet clinical requirements. Recently, researchers have focused their emphasis on phototherapy and immunogenic cell death (ICD) techniques. In response to the current problems of immunotherapy, a multifunctional drug delivery nanosystem (PDA-IMQ@CaCO3-blinatumomab, PICB) was constructed by using high physiological compatibility of polydopamine (PDA) and calcium carbonate (CaCO3). Toll-like receptor 7 (TLR7) agonist imiquimod (IMQ) and bispecific antibody (BsAb) blinatumomab were loaded onto PDA-CaCO3 nanoparticles (NPs). The findings revealed that the system exhibited the advantages of good dispersion, high stability, excellent physiological compatibility, low toxicity, and high drug loading rate. Compared to the control group, it resulted in a 2.4-fold decrease in FOXP3+ regulatory T-cells within the tumor and a 5.0-fold increase in CD4+ effector T-cells, and promoted the production of damage-related molecular patterns to reinvigorate the ICD effect. PICB had a strong inhibitory effect on tumor growth in 4T1 tumor-bearing mice, and has no toxicity to other organs. Therefore, the multifunctional drug delivery nanosystem constructed in this study could effectively exert the properties of various components in vivo, fully demonstrate the synergistic effect between immunotherapy and photothermal therapy, thus significantly improving the tumor therapeutic efficacy, and has a promising clinical application.
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To solve the problems existing in the clinical application of hypericin (Hyp) and tirapazamine (TPZ), a nano-drug delivery system with synergistic anti-tumor functions was constructed using mesoporous silica nanoparticles (MSN) and sodium alginate (SA). The system exhibited excellent stability, physiological compatibility and targeted drug release performance in tumor tissues. In the in vitro and in vivo experiments, Hyp released from MSN killed tumor cells through photodynamic therapy (PDT). The degree of hypoxia in the tumor tissue site was exacerbated, enabling TPZ to fully exert its anti-tumor activity. Our studies suggested that the synergistic effects between the components of the nano-drug delivery system significantly improve the anti-tumor properties of Hyp and TPZ.
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Alginatos , Antracenos , Nanopartículas , Perileno , Dióxido de Silicio , Tirapazamina , Microambiente Tumoral , Dióxido de Silicio/química , Microambiente Tumoral/efectos de los fármacos , Alginatos/química , Animales , Humanos , Tirapazamina/química , Tirapazamina/farmacología , Nanopartículas/química , Perileno/análogos & derivados , Perileno/química , Perileno/farmacología , Ratones , Antracenos/química , Línea Celular Tumoral , Fotoquimioterapia , Antineoplásicos/química , Antineoplásicos/farmacología , Antineoplásicos/uso terapéutico , Ácido Glucurónico/química , Ácidos Hexurónicos/química , Sistemas de Liberación de Medicamentos , Liberación de Fármacos , Porosidad , Fármacos Fotosensibilizantes/química , Fármacos Fotosensibilizantes/farmacología , Fármacos Fotosensibilizantes/uso terapéutico , Sinergismo Farmacológico , Sistema de Administración de Fármacos con Nanopartículas/químicaRESUMEN
The morbidity and death rates of calcified aortic valves|calcific aortic valve (CAV) disease (CAVD) remain high for its limited therapeutic choices. Here, we investigated the function, therapeutic potential, and putative mechanisms of Enoyl coenzyme A hydratase 1 (ECH1) in CAVD by various in vitro and in vivo experiments. Single-cell sequencing revealed that ECH1 was predominantly expressed in valve interstitial cells and was significantly reduced in CAVs. Overexpression of ECH1 reduced aortic valve calcification in ApoE-/- mice treated with high cholesterol diet, while ECH1 silencing had the reverse effect. We also identified Wnt5a, a noncanonical Wnt ligand, was also altered when ECH1 expression was modulated. Mechanistically, we found that ECH1 exerted anti-calcific actions through suppressing Wnt signaling, since CHIR99021, a Wnt agonist, may significantly lessen the protective impact of ECH1 overexpression on the development of valve calcification. ChIP and luciferase assays all showed that ECH1 overexpression prevented Runx2 binding to its downstream gene promoters (osteopontin and osteocalcin), while CHIR99021 neutralized this protective effect. Collectively, our findings reveal a previously unrecognized mechanism of ECH1-Wnt5a/Ca2+ regulation in CAVD, implying that targeting ECH1 may be a potential therapeutic strategy to prevent CAVD development.
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BACKGROUND: Rab1A not only regulates eukaryotic secretion, autophagy and intracellular traffic, but also extensively participates in the development of cancer. Thus, we collected data to investigate the clinical value of Rab1A in cancers. METHODS: English web database was searched for appropriate studies. The role of Rab1A in cancer patients was evaluated by combining hazard ratios and odds ratios. RESULTS: There were 15 studies in 14 articles, including 1,791 cancer patients. The results showed that upregulated Rab1A led to poor prognosis in cancer patients (pooled HR = 2.545, 95% CI = 1.924 - 3.367, p < 0.001). Notably, a high level of Rab1A was associated with a poorer prognosis than patients with a low level of Rab1A in digestive system cancer (pooled HR = 2.484, 95% CI = 1.796 - 3.437, p < 0.001). In order to explore the possible carcinogenic mechanism, we further analyzed and confirmed that high expression of Rab1A was associated with worse histologic grade, deeper tumor invasion, higher TNM stage, positive LN metastasis, positive neural invasion, positive vascular invasion, and larger tumor size (p < 0.05). CONCLUSIONS: Rab1A overexpression was associated with poor prognosis and adverse clinicopathological parameters in cancer patients and had the potential to be a target for future cancer therapy.
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Neoplasias del Sistema Digestivo , Humanos , Pronóstico , Modelos de Riesgos Proporcionales , Biomarcadores de Tumor/metabolismoRESUMEN
Background: Controversy exists regarding the advantages and risks of off-pump vs. on-pump coronary artery bypass grafting (CABG) for patients with diabetes. We therefore compare the early clinical outcomes of off-pump vs. on-pump procedures for diabetic patients with three-vessel disease. Materials and methods: We conducted a retrospective analysis of clinical data obtained from 548 diabetic patients with three-vessel coronary artery disease who underwent isolated CABG between January 2016 and June 2020. To adjust the differences of baseline characteristics between the off-pump CABG (OPCAB) and on-pump CABG (ONCAB) groups, propensity score matching (PSM) was used. Following 1:1 matching, we selected 187 pairs of patients for further comparison of outcomes within the first 30 days after surgery. Results: The preoperative characteristics of the patients between the two groups were clinically comparable after PSM. The OPCAB group exhibited a significantly higher incidence of incomplete revascularization (27.3% vs. 14.4%; P = 0.002) compared with the ONCAB group. No differences were seen in mortality within 30 days between the matched groups (1.1% vs. 3.7%; P = 0.174). Notably, the OPCAB group had a lower risk of respiratory failure or infection (2.1% vs. 7.0%; P = 0.025), less postoperative stroke (1.1% vs. 4.8%; P = 0.032), and reduced postoperative ventilator assistance time (35.8 ± 33.7 vs. 50.9 ± 64.8; P = 0.005). Conclusion: OPCAB in diabetic patients with three-vessel disease is a safe procedure with reduced early stroke and respiratory complications and similar mortality rate, myocardial infarction, and renal failure requiring dialysis to conventional on-pump revascularization.
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Left ventricular assist device in combination with clenbuterol has been demonstrated to significantly improve heart function in patients with advanced heart failure. However, the roles of clenbuterol in mechanical unloading and its underlying mechanism are poorly understood. A rat abdominal heart transplantation model has been developed to mimic mechanical unloading of the heart. The recipient rats were randomly segregated into experimental groups for the daily administration of either saline (the "Trans" group; n = 13) or clenbuterol (2 mg/kg, the "Trans + CB" group; n = 12). Another group of 10 rats served as a treatment mimic control/sham animals (the "Sham" group). All interventions were performed via intraperitoneal injections once daily for 4 weeks. The Trans group animals exhibited myocardial atrophy and dysfunction with decreased expression levels of transient receptor potential channel 3 (TRPC3) and phospholipase C-ß1 (PLC-ß1) at 4 weeks post-transplantation. Administration of clenbuterol improved cardiac function, prevented myocardial atrophy, and restored expression of TRPC3 and PLC-ß1 in the unloaded hearts of the "Trans + CB" animals at 4 weeks post-transplantation. Silencing of the TRPC3 gene by siRNA inhibited the pro-hypertrophic effect of clenbuterol in the rat primary cardiomyocytes in vitro. Furthermore, U73122, an inhibitor of the PLC-ß1/diacylglycerol (DAG) pathway, significantly attenuated clenbuterol-induced upregulation of TRPC3 in cardiomyocytes. These findings suggest that the anti-atrophic effect of clenbuterol may be dependent on the upregulation of TRPC3 through the activation of the PLC-ß1/DAG pathway during mechanical unloading. The results of our study reveal a potential target for the prevention and treatment of mechanical unloading-induced myocardial atrophy.
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Clenbuterol , Canales de Potencial de Receptor Transitorio , Humanos , Ratas , Animales , Clenbuterol/farmacología , Clenbuterol/metabolismo , Regulación hacia Arriba , Función Ventricular Izquierda/fisiología , Miocitos Cardíacos/metabolismo , Atrofia Muscular , Miocardio/patologíaRESUMEN
OBJECTIVE: To construct a prognostic model using artificial neural network (ANN) approach, providing an idea for the prediction and diagnosis of cholangiocarcinoma (CCA). STUDY DESIGN: Experimental study. Place and Duration of the Study: Department of General Surgery, Zhenjiang Hospital, Zhenjiang Province, China, between January and March 2022. METHODOLOGY: Available datasets were obtained from the Gene Expression Omnibus (GEO) database to construct the train cohort and the test cohort of CCA, and screened out the differentially expressed genes (DEGs) of CCA. Next, an ANN model for CCA diagnosis was constructed based on the scores of the DEGs and evaluated its accuracy and efficiency using ROC curves. Finally, the immune infiltration and the function of extracellular matrix (ECM) protein SPACRL1 were analysed to reveal the characteristic alterations in CCA. RESULTS: This analysis revealed 166 DEGs, mainly concentrated in the ECM organisation, neutrophil activation and other pathways. Then a set of 17 CCA disease signature genes scores were obtained to build an ANN prediction model and the ROC curve was plotted. The AUC in the train group (0.980) indicated that the accuracy of the diagnosis model is extremely high. Finally, there was a significant increase of B cells naïve (p=0.025), tregs (p=0.004), and macrophages M1 (p<0.001) in the tumour-microenvironment of CCA, while SPARCL1 was a protective factor on disease-specific survival (DSS) in CCA (p=0.009). CONCLUSION: This study has developed an accurate prediction model for CCA diagnosis, and identified SPARCL1 as pivotal factor in CCA by modulating the tumour immune-microenvironment. KEY WORDS: Cholangiocarcinoma, Artificial neural network, Immune microenvironment, Bioinformatics, Prognosis model, SPARCL1.
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Neoplasias de los Conductos Biliares , Colangiocarcinoma , Humanos , Bosques Aleatorios , Colangiocarcinoma/diagnóstico , Colangiocarcinoma/genética , Redes Neurales de la Computación , Conductos Biliares Intrahepáticos/patología , Neoplasias de los Conductos Biliares/diagnóstico , Neoplasias de los Conductos Biliares/genética , Microambiente TumoralRESUMEN
BACKGROUND: The sex-determining region Y-box 2 (SOX2) has been identified to be involved in tumor progression and prognosis in patients with gastric cancer (GC). However, its action is paradoxical. Thus, we conducted the first meta-analysis based on eligible studies to evaluate the clinical utility of SOX2 in GC only. METHODS: A thorough electronic search was performed to collect eligible studies. The hazard ratios (HRs) or odds ratios (ORs) with 95% confidence intervals (CIs) were generated from included studies to assess the strength of the association between SOX2 and prognosis and clinicopathological characteristics in GC. RESULTS: A total of 10 studies comprising 1321 patients with GC were identified for the meta-analysis. The pooled results revealed that high SOX2 expression was significantly associated with poor overall survival compared to low SOX2 expression (pooled HR = 1.485; 95% CI: 1.022-2.160; ð = .04). The statistical significance between SOX2 expression and overall survival was also established in univariate analysis (pooled HR = 1.606; 95% CI: 1.134-2.274; ð < .01), as well as recruitment time exceeding 2010 (pooled HR = 1.873; 95% CI: 1.041-3.371; ð = .04), follow-up time more than 5 years (pooled HR = 1.642; 95% CI: 1.066-2.527; ð = .02), and cutoff value of more than 5% of cells stained (pooled HR = 1.730; 95% CI: 1.162-2.577; ð < .01). Moreover, we verified that positive SOX2 expression was correlated with advanced tumor invasion depth (pooled OR = 0.494; 95% CI: 0.362-0.675; ð < .01) and positive vascular invasion (pooled OR = 1.515; 95% CI: 1.078-2.130; ð = .02). CONCLUSION: SOX2 could not only be an independent prognostic marker in GC but might also be a novel target for cancer therapy.
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Neoplasias Gástricas , Biomarcadores de Tumor/metabolismo , Humanos , Oportunidad Relativa , Pronóstico , Modelos de Riesgos Proporcionales , Factores de Transcripción SOXB1 , Neoplasias Gástricas/patologíaRESUMEN
A novel method for the preparation of antitumor drug vehicles has been optimized. Biological materials of chitosan oligosaccharide (CO) and γ-polyglutamic acid (γ-PGA) have previously been employed as modifiers to covalently modify graphene oxide (GO), which in turn loaded doxorubicin (DOX) to obtain a nano drug delivery systems of graphene oxide based composites (GO-CO-γ-PGA-DOX). The system was not equipped with the ability of initiative targeting, thus resulting into toxicity and side effects on normal tissues or organs. In order to further improve the targeting property of the system, the nucleic acid aptamer NH2 -AS1411 (APT) of targeted nucleolin (C23) was used to conjugate on GO-CO-γ-PGA to yield the targeted nano drug delivery system APT-GO-CO-γ-PGA. The structure, composition, dispersion, particle size and morphology properties of the synthesized complex have been studied using multiple characterization methods. Drug loading and release profile data showed that APT-GO-CO-γ-PGA is provided with high drug loading capacity and is capable of controlled and sustained release of DOX. Cell experimental results indicated that since C23 was overexpressed on the surface of Hela cells but not on the surface of Beas-2B cells, APT-GO-CO-γ-PGA-DOX can target Hela cells and make increase toxicity to Hela cells than Beas-2B cells, and the IC50 value of APT-GO-CO-γ-PGA-DOX was 3.23±0.04â µg/mL. All results proved that APT-GO-CO-γ-PGA can deliver antitumor drugs in a targeted manner, and achieve the effect of reducing poison, which indicated that the targeted carrier exhibits a broad application prospect in the field of biomedicine.
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Antineoplásicos/farmacología , Aptámeros de Nucleótidos/química , Doxorrubicina/farmacología , Portadores de Fármacos/química , Grafito/química , Nanocompuestos/química , Oligodesoxirribonucleótidos/química , Aptámeros de Nucleótidos/toxicidad , Quitina/análogos & derivados , Quitina/química , Quitina/toxicidad , Quitosano , Portadores de Fármacos/toxicidad , Liberación de Fármacos , Grafito/toxicidad , Células HeLa , Humanos , Ácidos Nucleicos Inmovilizados/química , Ácidos Nucleicos Inmovilizados/toxicidad , Nanocompuestos/toxicidad , Oligodesoxirribonucleótidos/toxicidad , Oligosacáridos , Ácido Poliglutámico/análogos & derivados , Ácido Poliglutámico/química , Ácido Poliglutámico/toxicidadRESUMEN
OBJECTIVE: It is unclear whether nucleophosmin (NPM) participates in cardiovascular disease. The present study aimed to investigate the role and underlying mechanisms of NPM in atherosclerosis. METHODS: Levels and location of NPM in human carotid atherosclerotic plaques and healthy controls were detected by real-time polymerase chain reaction, immunoblots, and immunofluorescence. Atherosclerotic prone ApoE-/- mice were fed with a Western diet for 16 weeks as an in vivo model. Human primary umbilical vein endothelial cells (HUVECs) were cultured as an in vitro model. RESULTS: Compared with controls, we found that NPM levels in human carotid atherosclerotic plaques were more than twice as high as in normal arteries, which mainly localized in endothelial cells. In vivo, adenovirus-containing NPM small hairpin RNA attenuated atherosclerotic lesion and promoted plaque stabilization in ApoE-/- mice fed a Western diet by reducing vascular inflammation, maintaining endothelial function, and decreasing macrophage infiltration. Furthermore, NPM knockdown decreased nuclear factor-κB (NF-κB) p65 phosphorylation. In cultured HUVECs, palmitic acid increased the protein levels of NPM and induced the expression of inflammatory cytokines and monocyte adhesion, whereas NPM knockdown attenuated this effect. In HUVECs, NPM protein physically interacted with NF-κB p65 subunit and promoted its nuclear transposition. NPM also increased the transcriptional activity of NF-κB p65 promoter and enhance its binding to target genes, including interleukin-1ß, interleukin-6, intercellular adhesion molecule-1, and E-selectin. CONCLUSIONS: These data provide novel evidence that NPM promotes atherosclerosis by inducing vascular inflammation and endothelial dysfunction through the NF-κB signaling pathway and suggest that NPM may be a promising target for atherosclerosis prevention and treatment.
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Aterosclerosis/metabolismo , Enfermedades de las Arterias Carótidas/metabolismo , Endotelio Vascular/metabolismo , Proteínas Nucleares/metabolismo , Placa Aterosclerótica/metabolismo , Vasculitis/metabolismo , Animales , Aterosclerosis/etiología , Enfermedades de las Arterias Carótidas/etiología , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Células Endoteliales de la Vena Umbilical Humana , Humanos , Inflamación/etiología , Inflamación/metabolismo , Ratones , FN-kappa B/metabolismo , Proteínas Nucleares/efectos adversos , Proteínas Nucleares/análisis , Nucleofosmina , Transducción de Señal , Vasculitis/etiologíaRESUMEN
BACKGROUND: Graft rejection continues to be a major barrier to long-term engraftment after transplantation. Autophagy plays an important role in cardiac injury pathogenesis. The bromodomain and extraterminal protein inhibitor (S)-tert-butyl2-(4-(4-chlorophenyl)-2,3,9-trimethyl-6H-thieno[3,2-f][1,2,4]triazolo[4,3-a][1,4]diazepin-6-yl)acetate (JQ1) inhibits inflammatory responses. However, the beneficial effect of JQ1 on transplant and the potential role of autophagy in the protective effect of graft survival are yet to be investigated. METHODS: Syngeneic or allogeneic heterotopic heart transplantation was performed using C57BL/6 or BALB/c donors for C57BL/6 recipients through different treatments. Some mice were used to observe the survival of the grafts. The other mice were euthanized on the third, fifth, and seventh days after surgery. The graft samples were taken for cytokines and autophagy pathway analyses. RESULTS: Our study revealed that JQ1 treatment prolonged cardiac allograft survival. JQ1 increased the expression levels of liver kinase beta 1, autophagy-specific gene 5, and microtubule-associated protein light chain3-II (LC3-II) and potentiated the phosphorylation of AMP-activated protein kinase, unc-51-like kinase 1 (ULK1), and autophagy-specific gene 14 in allografts. A conditional autophagy-specific gene 5 deletion donor was utilized to abrogate the effect induced by JQ1. The combined use of JQ1 with bafilomycin A1 partially reversed the effect of JQ1, suggesting that autophagy is involved in the signaling pathway in graft survival. JQ1 downregulated the expression of inflammatory cytokines, such as interleukin-6, interleukin-1ß, tumor necrosis factor-α, and interferon-γ, which was abrogated when autophagy was inhibited. CONCLUSIONS: JQ1 prolonged cardiac allograft survival by potentiating myocardial autophagy through the liver kinase beta 1-AMP-activated protein kinase-ULK1 signaling pathway and inhibiting the subsequent release of inflammatory cytokines. This result might provide novel insights for extending transplant survival.
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Autofagia/efectos de los fármacos , Azepinas/farmacología , Supervivencia de Injerto/efectos de los fármacos , Trasplante de Corazón , Miocitos Cardíacos/efectos de los fármacos , Proteínas/antagonistas & inhibidores , Triazoles/farmacología , Proteínas Quinasas Activadas por AMP/metabolismo , Animales , Proteína 5 Relacionada con la Autofagia/genética , Proteína 5 Relacionada con la Autofagia/metabolismo , Homólogo de la Proteína 1 Relacionada con la Autofagia/metabolismo , Azepinas/química , Citocinas/metabolismo , Trasplante de Corazón/efectos adversos , Mediadores de Inflamación/metabolismo , Masculino , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Miocitos Cardíacos/metabolismo , Miocitos Cardíacos/patología , Miocitos Cardíacos/trasplante , Dominios Proteicos , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas/metabolismo , Transducción de Señal , Factores de Tiempo , Triazoles/químicaRESUMEN
N-heterocyclic carbenes-modified half-sandwich iridium(III) complex [(η5-C5Me4C6H4C6H5)Ir(C^C)Cl]PF6 (C1) (where C^C is a N-heterocyclic carbene ligand) can effectively prevent the proliferation of human cervical cancer cells. Here, this study aims to investigate the in-deep anticancer effects of this complex on non-small cell lung cancer cells and explore the underlying molecular mechanism. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay showed that iridium(III) complex had potent cytotoxicity studies towards non-small cell lung cancer cells (A549), human lung squamous cells (L78), human cervical cancer cells (Hela) and human bronchial epithelial cells (BEAS-2B). Colocalization and cellular uptake studies were analyzed by confocal microscopy. Notably, C1 targeted lysosomes and entered the cancer cells partially through an energy-dependent pathway, inducing the release of cathepsins and other proteins. These proteins regulated lysosomal-mitochondrial dysfunction, thus leading to the release of cytochrome c (cyt c), which amplified apoptotic signals by activating many downstream pathways such as caspase pathways to promote cell apoptosis. The results showed that the inhibitory mechanism of this organometallic iridium(III) complex may involve caspase-associated apoptosis initiated by the lysosomal-mitochondrial pathway.
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Antineoplásicos/farmacología , Iridio/química , Iridio/farmacología , Lisosomas/metabolismo , Metano/análogos & derivados , Compuestos Organometálicos/farmacología , Células A549 , Antineoplásicos/química , Apoptosis/efectos de los fármacos , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Complejos de Coordinación/química , Complejos de Coordinación/farmacología , Citocromos c/metabolismo , Ensayos de Selección de Medicamentos Antitumorales , Células HeLa , Compuestos Heterocíclicos/química , Compuestos Heterocíclicos/farmacología , Humanos , Neoplasias Pulmonares/metabolismo , Lisosomas/efectos de los fármacos , Metano/química , Microscopía Confocal , Mitocondrias/metabolismo , Compuestos Organometálicos/químicaRESUMEN
Browning of white adipose tissue (WAT) has been recognized as an important strategy for the treatment of obesity, insulin resistance, and diabetes. Enoyl coenzyme A hydratase 1 (ECH1) is a widely known enzyme involved in lipid metabolism. However, whether and how ECH1 is implicated in browning of WAT remain obscure. Adeno-associated, virus-mediated genetic engineering of ECH1 in adipose tissue was used in investigations in mouse models of obesity induced by a high-fat diet (HFD) or browning induced by cold exposure. Metabolic parameters showed that ECH1 overexpression decreased weight gain and improved insulin sensitivity and lipid profile after 8 wk of an HFD. Further work revealed that these changes were associated with enhanced energy expenditure and increased appearance of brown-like adipocytes in inguinal WAT, as verified by a remarkable increase in uncoupling protein 1 and thermogenic gene expression. In vitro, ECH1 induced brown fat-related gene expression in adipocytes differentiated from primary stromal vascular fractions, whereas knockdown of ECH1 reversed this effect. Mechanistically, ECH1 regulated the thermogenic program by inhibiting mammalian target of rapamycin signaling, which may partially explain the potential mechanism for ECH1 regulating adipose browning. In summary, ECH1 may participate in the pathology of obesity by regulating browning of WAT, which probably provides us with a new therapeutic strategy for combating obesity.
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Tejido Adiposo Pardo/enzimología , Isomerasas de Doble Vínculo Carbono-Carbono/genética , Isomerasas de Doble Vínculo Carbono-Carbono/metabolismo , Terapia Genética/métodos , Enfermedades Metabólicas/terapia , Obesidad/terapia , Tejido Adiposo Pardo/crecimiento & desarrollo , Tejido Adiposo Blanco/enzimología , Tejido Adiposo Blanco/crecimiento & desarrollo , Animales , Frío , Dieta Alta en Grasa , Metabolismo Energético , Ingeniería Genética , Resistencia a la Insulina , Masculino , Ratones , Ratones Endogámicos C57BL , Serina-Treonina Quinasas TOR/metabolismo , Termogénesis , Aumento de PesoRESUMEN
BACKGROUND & AIM: Panax notoginseng saponins are believed to promote wound healing due to its anti-proliferative effect on fibroblasts. The present work was therefore aimed to examine the beneficial effect of PNS on wound healing in vitro and in a murine model of cutaneous wound. METHODS: The in vitro effects of Panax notoginseng saponins on the proliferation of and nitric oxide (NO) synthesis in human fibroblast 3T3 cells were studied. The in vivo effects of Panax notoginseng saponins were examined in C57 mice with dorsal cutaneous wound. The healing rate and scar formation were followed after treatment with Panax notoginseng saponins. The histology and fibroblast accumulation in the wounds were studied using hematoxylin and eosin (H&E) staining. Expression of α-smooth muscle actin (α-SMA) was examined by immunohistochemistry. RESULTS: Panax notoginseng saponins inhibited the proliferation of human fibroblast 3T3 with an EC50 of 1.825 mM Panax notoginseng saponins (0.1 mM) significantly promoted NO production (P < 0.01) and NO synthase activity (P < 0.01) of 3T3. In C57 mice with dorsal cutaneous wounds, 0.1 mM Panax notoginseng saponins significant expedited wound healing by reducing the size of lesions and suppressing the formation of scar. H&E staining revealed that treatment with Panax notoginseng saponins suppressed fibroblast accumulation in wound areas, while immunohistochemistry showed a significant reduction in α-SMA expression by 0.1 mM Panax notoginseng saponins. CONCLUSION: Panax notoginseng saponins are a promising drug candidate that can accelerate wound healing and reduce scar formation.
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Cicatriz/prevención & control , Panax notoginseng/química , Saponinas/administración & dosificación , Piel/efectos de los fármacos , Cicatrización de Heridas/efectos de los fármacos , Células 3T3 , Actinas/metabolismo , Animales , Proliferación Celular/efectos de los fármacos , Cicatriz/patología , Modelos Animales de Enfermedad , Humanos , Masculino , Ratones , Óxido Nítrico/metabolismo , Transducción de Señal/efectos de los fármacos , Piel/citología , Piel/lesiones , Piel/patologíaRESUMEN
Background and aims: Biological mechanisms that control liver regeneration remain poorly defined. However, these mechanisms are remarkable issues in the clinic that affect management of hepatic loss caused by liver surgery, traumatic injury, chronic infection, or liver poisoning. Increasing evidence has shown that various growth factors, cytokines, and metabolic signaling pathways affect the liver regenerative process. Our aim is to study the effect of bromodomain and extraterminal (BET) protein inhibition on liver regeneration and its mechanism. Methods: We studied the role of BET protein inhibitor, JQ1, in liver regeneration in a mouse model after 70% partial hepatectomy (PH). We evaluated yes-associated protein (YAP)/transcriptional co-activator with PDZ-binding motif (TAZ) and Notch signaling pathways, which were affected by BET protein inhibitor in mouse hepatic tissues and primary hepatocytes in vivo and AML12 cell lines in vitro. We evaluated the relationship of YAP/TAZ and Notch signaling pathway using YAP/TAZ pathway inhibitor in liver regeneration in vivo. Moreover, we analyzed the relationship of YAP/TAZ and Notch signaling pathways via overexpression or RNA silencing of Yap in AML12 cells. Furthermore, we used Yap overexpression mouse model to examine whether it can rescue liver regeneration damage caused by inhibition of BET proteins. Results: In this study, we report that BET protein inhibitor JQ1 molecule impairs the early phase of liver regeneration in a mouse model after 70% PH. Mechanistically, YAP/TAZ and Notch1-NICD pathways were suppressed by BET protein inhibitor in mouse hepatic tissues and primary hepatocytes in vivo and mouse AML12 cell lines in vitro. By using YAP/TAZ pathway inhibitor, we confirmed that the liver regeneration and the activation of Notch pathway were impaired by the inhibition of YAP/TAZ pathway in vivo. Furthermore, the study showed that Yap knockdown by shRNA in normal mouse hepatic cell line downregulated Notch1 signal transduction, whereas Yap overexpression promoted Notch1-NICD signals. Specific overexpression of Yap in mouse liver could rescue the effect of BET protein inhibition on liver regeneration injury. Conclusion: These results revealed the crucial role of the YAP/TAZ-Notch1-NICD axis in liver regeneration. Therefore, BET protein inhibitors must be used in caution in the treatment of hepatic diseases by reason of its suppressive roles in liver regeneration.
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Proteínas Adaptadoras Transductoras de Señales/metabolismo , Proteínas de Ciclo Celular/metabolismo , Regeneración Hepática , Receptor Notch1/química , Receptor Notch1/metabolismo , Transducción de Señal , Transactivadores/metabolismo , Animales , Proliferación Celular , Regulación hacia Abajo , Hepatectomía , Hepatocitos/metabolismo , Hígado/metabolismo , Masculino , Ratones Endogámicos C57BL , Proteínas del Tejido Nervioso/antagonistas & inhibidores , Proteínas del Tejido Nervioso/metabolismo , Dominios Proteicos , Receptores de Superficie Celular/antagonistas & inhibidores , Receptores de Superficie Celular/metabolismo , Proteínas Señalizadoras YAPRESUMEN
Background: The liver coordinates a series of metabolic adaptations to maintain the energy balance of the system and provide adequate nutrients to key organs, tissues and cells during starvation. However, the mediators and underlying molecular mechanisms that mediate these fasting-induced adaptive responses remain unclear. Materials and methods: Male wild-type C57BL/6J littermates (8-weeks-old) were intraperitoneally injected with MCC950 or vehicle, and then randomly divided into three groups: fed, fasted, and refed. Plasma IL1ß and insulin levels were detected by ELISA kits. Plasma and hepatic metabolites were determined using commercial assay kits. HepaRG cell line was applied to verify the regulation of NLRP3 on lipogenesis. Results: NOD-like receptor protein 3 (NLRP3) and its downstream inflammatory cytokines were significantly suppressed after 24 h fasting and recovered upon 6 h refeeding in plasma and liver tissues of mice. Moreover, fasting-induced hepatic steatosis and accompanied liver injury were ameliorated when mice were intraperitoneally injected with MCC950 (a selective NLRP3 inhibitor). Further study revealed that MCC950 suppressed sterol regulatory element-binding protein-1c (SREBP-1c) expression and transcriptional activity, thus inhibited lipogenesis in the liver, which may explain its role in stabilizing lipid metabolism. Conclusion: The NLRP3 inhibitor-MCC950 protects against fasting-induced hepatic steatosis. The novel and critical role of NLRP3 in lipogenesis may explain its importance in regulating the adaptive responses of the liver upon starvation stress and may provide therapeutic value.
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ETHNOPHARMACOLOGICAL RELEVANCE: Qiwei Tiexie capsule (QWTX) is a representative prescription of Tibetan medicine, which is widely used for long-term treatment of chronic liver disease and nonalcoholic fatty liver disease (NAFLD). AIM OF THE STUDY: This study explored the effects and mechanism of QWTX on 3T3-L1 adipocytes and NAFLD. MATERIALS AND METHODS: The 3T3-L1 preadipocytes and NAFLD rat model were used in the study. In 3T3-L1 cells, the cytotoxicity of QWTX was tested by CKK-8, and glucose uptake and fat acid oxidation were assessed by 2-deoxy-D-[3H] glucose and [1-14C] palmitic acid, respectively. The expression levels of carnitine palmitoyltransferase-1 (CPT-1), liver X receptor α (LXRα), peroxisome proliferator-activated receptor (PPAR) γ, inducible nitric oxide synthase (iNOS), ikappa B α (IκBα), and AKT were determined by PCR and western blot. NAFLD was established by the administration of fat emulsion and sucrose for 9 weeks. The effects of QWTX on lipid metabolism, liver function, and hepatic morphology were observed in NAFLD rats by HE and transmission electron microscope. Serum level of nitric oxide (NO) and fee fatty acid (FFA), superoxide dismutase (SOD) and malondialdehyde (MDA) contents in the liver, as well as the expression levels of Cytochrome P450 2E1 (CYP2E1), NF-κB, monocyte chemoattractant protein 1 (MCP-1), CPT-1, LXRα, PPARα, PPARß/δ, PPARγ, and iNOS were all detected. RESULTS: QWTX showed no cell cytotoxicity in 3T3-L1 preadipocyte cells, and increased the 14CO2 production rate to 4.15, which indicated the reducing the fatty accumulation. In NAFLD, QWTX attenuated liver steatosis, fat vacuoles and inflammation from the HE staining and electron micrograph tests. For the oxidative stress biomarkers, serum FFA level was reduced and serum NO level was enhanced after QWTX treatment. In liver tissue, SOD was decreased and MDA was significantly increased in NAFLD, and both of them were restored by QWTX. NF-κB and CYP2E1 were also upregulated in NAFLD, while downregulated by QWTX. Downregulation of LXRα, PPARγ and iNOS by QWTX were both observed in the 3T3-L1 adipocytes and NAFLD model. CONCLUSIONS: QWTX protected the liver injury in differentiated 3T3-L1 adipocytes and NAFLD by regulating the LXRα, PPARγ, and NF-κB-iNOS-NO signal pathways.
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Medicamentos Herbarios Chinos/farmacología , Enfermedad del Hígado Graso no Alcohólico/tratamiento farmacológico , Sustancias Protectoras/farmacología , Transducción de Señal/efectos de los fármacos , Células 3T3-L1 , Animales , Cápsulas , Citocromo P-450 CYP2E1/metabolismo , Dieta Alta en Grasa/efectos adversos , Modelos Animales de Enfermedad , Medicamentos Herbarios Chinos/uso terapéutico , Humanos , Hígado/efectos de los fármacos , Hígado/metabolismo , Hígado/patología , Receptores X del Hígado/metabolismo , Masculino , Medicina Tradicional Tibetana , Ratones , FN-kappa B/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Enfermedad del Hígado Graso no Alcohólico/etiología , Enfermedad del Hígado Graso no Alcohólico/patología , PPAR gamma/metabolismo , Sustancias Protectoras/uso terapéutico , Ratas , Ratas Sprague-Dawley , Pruebas de Toxicidad , Resultado del TratamientoRESUMEN
BACKGROUND: Resistance of tolerance induction in sensitized transplantation is mainly caused by generation of memory T cells. It is unknown whether alteration of graft niche such as level of pro-inflammatory cytokines can affect generation of memory T cells. METHODS: IL-6 deficient or wild-type (WT) C57BL/6 heart grafts were transplanted into pre-sensitized wild-type BALB/c recipients. Frequencies of memory T cells in the peripheral blood, grafts, and spleen were evaluated. RESULTS: We revealed that deficiency of donor IL-6 could significant prolong sensitized allograft survival. Compared with counterpart of WT group, frequency of effector memory CD4â¯+â¯T cells (CD4â¯+â¯CD44â¯+â¯CD62L-) in the peripheral blood was significantly lower in the IL-6 KO group (pâ¯=â¯.026) at day 3 post-transplantation. Frequency of effector memory CD8â¯+â¯T cells (CD8â¯+â¯CD44â¯+â¯CD62L-) in the peripheral blood was significantly lower in the IL-6 KO group (pâ¯<â¯.0001) at day 3 post-transplant in comparison to that of WT group. No significant difference of central memory T cells was found between these groups. Histology demonstrated that deficiency of donor pro-inflammatory cytokine IL-6 (IL-6 KO group) preserved cardiac architecture with a mild infiltration of lymphocytes, whereas wild-type donor (control group) caused an evident lymphocytic infiltration within myocardial fibers of grafts and destruction of cardiac structure. CONCLUSION: Deficiency of pro-inflammatory IL-6 of donor graft could effectively prolong sensitized allograft survival, which was caused by a remarkable decrease of peripheral memory T cells rather than central memory T cells. This unveiled mechanism of targeting IL-6 signaling pathway might provide a novel insight into preventing allograft rejection for sensitized transplant recipients.
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Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Trasplante de Corazón , Interleucina-6/metabolismo , Miocardio/inmunología , Trasplante de Piel , Animales , Células Cultivadas , Modelos Animales de Enfermedad , Humanos , Inmunización , Memoria Inmunológica , Interleucina-6/genética , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Trasplante HomólogoRESUMEN
Two-dimensional (2D) gold nanoparticle (Au NP) monolayer film possesses a lot of fascinating peculiarities, and has shown promising applications in photoelectrical devices, catalysis, spectroscopy, sensors, and anticounterfeiting. Because of the localized surface plasmon resonance (LSPR) property predetermined by the natural structure of metal nanoparticles, it is usually difficult to realize the reversible LSPR transition of 2D film. In this work, we report on the fabrication of a large-area free-standing Au NP monolayer film with dual-responsive switchable plasmonic property using a pH- or thermal-responsive dendronized copolymer as a stimuli-sensitive linker. In this system, an oligoethylene-glycol-based (OEG-based) dendronized copolymer (named PG1A) with pH or temperature sensitivity was first modified onto the surface of a Au NP. Then, polyethylene glycol dibenzyl aldehyde (PEG-DA) was introduced to interact with the amino moieties from PG1A before the process of oil-water interfacial self-assembly of NPs, resulting in an elastic, robust, pH- or temperature-sensitive interpenetrating network among Au NPs in monolayer films. In addition, the film could exhibit reversibly plasmonic shifts of about 77 nm and inherent color changes through varying temperature or pH. The obtained free-standing monolayer film also shows an excellent transferable property, which can be easily transferred onto substrates such as plastic molds, PDMS, copper grids, and silicon wafers. In virtue of these peculiarities of the free-standing property, special plasmonic signal, and homologous macroscopic color, the transferred film was primely applied to an anticounterfeiting security label with clear color change at the designed spots, providing a new avenue to plasmonic nanodevices with various applications.
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The bromo- and extra-terminal domain (BET) signaling pathway plays an important role in cell proliferation, immune responses, and pro-inflammatory events. JQ1 as a first-in-class potent and selective inhibitor of the BRD4 signaling pathway is widely used for tumor biology studies. It was found that JQ1 could potently reduce cancer cell viability in vitro and in vivo. The underlying mechanisms include an effect on cell cycle arrest in the G1 phase and a decrease in the percentage of cells in the S phase. Furthermore, JQ1 could alter cytokines expressions not only in T cells but also in dendritic cells (DCs). Apoptosis of tumor cells was induced by JQ1 through downregulation of E2f1 protein expression. In addition, JQ1 exhibited a potent suppressive effect on ERα and androgen receptor (AR) signaling pathways in breast and prostate cancers. Accumulating evidence supports the notion of BRD4 suppression as a target of therapeutic intervention in clinical oncology. Our present review article advances the understanding of the role of the JQ1 / BRD4 protein.