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This study explores the intersection of creative travel and flow experiences among foreign students, a topic not extensively explored in tourism research. Specifically, it examines the mediating role of flow experiences in the relationship between students' novelty-seeking behaviors and their intention to engage in creative travel. Additionally, the research investigates how familiarity with a destination moderates this relationship. Employing structural equation modeling, the study analyzes data from 704 Chinese students in Thailand. The findings reveal that flow experiences positively mediate the link between the students' pursuit of novelty and their creative travel intentions. Moreover, the extent of familiarity with the destination was found to modify the relationship between novelty seeking and flow experiences. This research contributes to the theoretical understanding of these dynamics and offers practical insights for stakeholders in creative travel marketing and management.
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RATIONALE: Anti-CD19-targeted chimeric antigen receptor (CAR) T cell therapy is effective in treating relapsed/refractory diffuse large B-cell lymphoma (DLBCL). This therapy is associated with several side effects that can be life-threatening such as cytokine release syndrome (CRS). However, chylothorax associated with CRS after CAR-T therapy has not been reported. PATIENT CONCERNS: A 23-year-old male diagnosed with DLBCL relapsing after autologous peripheral blood stem cell transplantation was treated with anti-CD19-targeted CAR-T cell therapy. After CAR-T cell transfusion, he developed grade 3 CRS includes fever, dyspnea, tachycardia and hypotension. The symptoms of CRS persisted and chest plain film revealed bilateral pleural effusion. DIAGNOSIS: Chylothorax was confirmed by the pleural effusion analysis that triglyceride level was 1061 mg/dL. Bacterial and fungal culture of pleural fluid reported no pathogen was detected. Cytological examination of pleural effusion revealed no malignant cells. INTERVENTIONS: The chylothorax resolved after treatment with intravenous administration of tocilizumab. OUTCOMES: On 30-day follow-up, the patient was in stable clinical condition with complete remission of DLBCL on whole-body positron emission tomography scan. LESSONS: We reported a rare case of CAR-T associated chylothorax in a patient with relapsed and refractory DLBCL. Grade 3 CRS with high interleukin-6 level was presented in our patient. The symptoms of CRS were improved with tocilizumab treatment and the chylothorax resolved later on. It is suggested that high interleukin-6 releases might induce chyle leakage resulting from activations of endothelium and coagulation. Our finding highlights the occurrence of chylothorax during the course of CAR-T cell therapy and the importance of proper monitoring and prompt management of this life-threatening side effect.
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Quilotórax , Linfoma de Células B Grandes Difuso , Trasplante de Células Madre de Sangre Periférica , Derrame Pleural , Receptores Quiméricos de Antígenos , Masculino , Humanos , Adulto Joven , Adulto , Receptores Quiméricos de Antígenos/uso terapéutico , Receptores de Antígenos de Linfocitos T , Quilotórax/etiología , Quilotórax/terapia , Interleucina-6/uso terapéutico , Recurrencia Local de Neoplasia/tratamiento farmacológico , Linfoma de Células B Grandes Difuso/tratamiento farmacológico , Inmunoterapia Adoptiva/efectos adversos , Inmunoterapia Adoptiva/métodos , Síndrome de Liberación de Citoquinas/tratamiento farmacológico , Antígenos CD19 , Derrame Pleural/tratamiento farmacológicoRESUMEN
OBJECTIVE: Estrogen has the potential to influence brain physiology implicated in dementia pathogenesis. Hormone replacement therapy (HRT) might be expected to influence the risk of dementia. Observational data indicated that HRT was associated with reductions in dementia risk, but experimental evidence demonstrates that HRT increases the incidence of dementia. To determine the effect of HRT on risk of dementia, a retrospective cohort study was performed using a nationwide claims dataset in Taiwan. METHODS: A population-base longitudinal study was performed using data from the Longitudinal Health Insurance Database in Taiwan. A total of 35,024 women with HRT were enrolled as the exposed cohort and 70,048 women without HRT were selected on the basis of propensity matching as the comparison cohort. All of the subjects were followed up until the diagnosis of dementia, death, or at the end of December 31, 2013, whichever occurred first. Overall, the average duration of follow-up (±SD) in the HT and comparison cohort was 12.3(±2.3) and 12.2 (±2.4), respectively. The Cox proportional hazards regression models were conducted to produce hazard ratios (HRs) with 95% confidence intervals (CIs) to evaluate the association of HRT with the risk of dementia. RESULTS: In the follow-up period, the cumulative incidence of dementia for the HRT cohort (20.04 per 1,000) was significantly higher than the corresponding cumulative incidence for the comparison cohort (15.79 per 1,000), resulting in an adjusted hazard ratio of 1.35 (95% CI, 1.13-2.62). There was an increased risk of dementia with a higher cumulative dose of HRT prescription (p for trend <0.0001). CONCLUSION: This cohort study documented that HRT was associated with an increased risk of dementia. The clinical implications of this study merit further investigations.
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Oxaliplatin, a third-generation platinum derivative, has become one of the main chemotherapeutic treatments for esophagus, gastric and colorectal cancer; however, it is still unclear the potential effectiveness for pancreatic ductal adenocarcinoma (PDAC) with gemcitabine resistance. Here, we observed that PDAC tumors have low level of organic cation transporter 2 (OCT2, also known as SLC22A2) compared with non-tumor tissues and identified that OCT2 expression is positively correlated with oxaliplatin sensitivity in PDAC cells. Treatment of OCT2 inhibitors or knockdown of OCT2 expression significantly decreased the sensitivity to oxaliplatin in PANC-1 cells. In addition, bisulfite sequencing polymerase chain reaction analysis revealed that higher methylation frequency represses OCT2 expression in gemcitabine-resistant PANC-1 (PANC-1/GR) cells. Moreover, we found that treatment of DNA methyltransferase (DNMT) inhibitors, decitabine or 5-azacytidine recover OCT2 expression and oxaliplatin sensitivity in PANC-1/GR cells, and DNMT1 level has inverse correlation with OCT2 expression in PDAC cells and tumors. Our findings jointly suggest that OCT2 expression is a potential and predictive marker for evaluating oxaliplatin sensitivity and developing alternative treatments for PDAC patients with gemcitabine resistance.
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Carcinoma Ductal Pancreático , Neoplasias Pancreáticas , Carcinoma Ductal Pancreático/patología , Línea Celular Tumoral , Humanos , Transportador 2 de Cátion Orgánico/metabolismo , Oxaliplatino/farmacología , Oxaliplatino/uso terapéutico , Neoplasias Pancreáticas/patología , Neoplasias PancreáticasRESUMEN
The entrapment of peripheral nerves is associated with chronic neuroinflammation and neuropathic pain, and perineural injection therapy with glucose is emerging as an effective treatment for peripheral entrapment neuropathy. However, the mechanism underlying the pharmacological effect of glucose on nerves remains unclear. One of the hypothesized mechanisms is that glucose reduces neurogenic inflammation. Therefore, we investigated the effects of high glucose concentrations on cytokine-induced neuroinflammation in vitro. Human SH-SY5Y neuronal cells were challenged with 10 ng/mL TNF-α for 16 h and subsequently treated with different glucose concentrations (0-25 mM) for 24 h. Cell viability was evaluated using the diphenyltetrazolium bromide assay, and proinflammatory cytokine levels were assessed using ELISA and quantitative PCR. In addition, mRNA levels of NF-κB and cyclooxygenase-2 were analyzed using quantitative PCR. Exposure to 10 ng/mL TNF-α resulted in decreased viability of SH-SY5Y cells and significant upregulation of IL-6, IL-1ß, NF-κB, and cyclooxygenase-2. Subsequent exposure to high glucose levels (25 mM) markedly reduced the upregulation of IL-6, IL-1ß, cyclooxygenase-2, and NF-κB, and restored the functional metabolism of SH-SY5Y cells, compared with that of the normal glucose control. Our findings suggest that high glucose concentrations can mitigate TNF-α-induced NF-κB activation, upregulation of proinflammatory cytokines, and metabolic dysfunction.
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Background: Sesamin is a major bioactive compound in sesame seeds and has various biological properties, including anti-inflammatory and anticancer activities. Here, we explored whether sesamin activates p53, which is widely inhibited in cervical cancer cells, thereby inducing p53-mediated apoptosis. Methods: Human HeLa and SiHa cervical cancer cells and normal Hs68 dermal cells were used as cell models. Cell proliferation, cell cycle distribution, and apoptosis were evaluated by the CCK-8 assay and flow cytometry using PI/Annexin V staining, respectively. Protein expression and phosphorylation were determined using western blotting. The involvement of p53 in the apoptotic cascade was assessed by a specific inhibitor. Results: Sesamin (75 and 150 µM) clearly inhibited SiHa and HeLa cell proliferation in a dose-dependent fashion, but did not affect the proliferation of Hs68 cells. Meanwhile, sesamin increased the sub-G1 phase ratio and apoptosis, up to approximately 38.5% and 37.8%, respectively. Furthermore, sesamin induced p53 phosphorylation at serine-46 and serine-15 and upregulated the levels of PUMA, Bax, and PTEN, while inhibiting AKT phosphorylation at serine-473. Inhibition of p53 by pifithrin-α significantly reduced the levels of PUMA, Bax, and PTEN but restored AKT phosphorylation in SiHa cells exposed to sesamin. Pifithrin-α also reduced apoptosis and restored the proliferation of HeLa and SiHa cells exposed to sesamin. Conclusions: These findings indicate that sesamin inhibits cervical cancer cell proliferation, and its mechanism may be attributed to the induction of p53/PTEN-mediated apoptosis. This suggests that sesamin might be useful as an adjuvant in promoting anti-cervical cancer treatments.
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Apoptosis/efectos de los fármacos , Dioxoles/farmacología , Lignanos/farmacología , Neoplasias del Cuello Uterino/tratamiento farmacológico , Ciclo Celular , Proliferación Celular/efectos de los fármacos , Dioxoles/uso terapéutico , Ensayos de Selección de Medicamentos Antitumorales , Femenino , Células HeLa , Humanos , Lignanos/uso terapéutico , Fosfohidrolasa PTEN/metabolismo , Transducción de Señal/efectos de los fármacos , Proteína p53 Supresora de Tumor/metabolismo , Neoplasias del Cuello Uterino/patologíaRESUMEN
BACKGROUND: Adenine is a purine with a role in cellular respiration and protein synthesis. It is considered for its pharmacological potential. We investigated whether anti-inflammatory effect of adenine benefits on the proliferation and maturation of osteoblastic cells. METHODS: Human osteoblast-like cells (MG-63) were cultured with adenine under control conditions or pre-treated with 10ng/mL of tumor necrosis factor-α (TNF-α) followed by adenine treatment. Cell viability was examined using dimethylthiazol diphenyltetrazolium bromide (MTT) assay. Expression of cytokines and osteogenic markers were analyzed using quantitative PCR (qPCR) and ELISA. Enzyme activity of alkaline phosphatase (ALP) and collagen content were measured. RESULTS: TNF-α exposure led to a decreased viability of osteoblastic cells. Treatment with adenine suppressed TNF-α-induced elevation in IL-6 expression and nitrite oxide production in MG-63 cells. Adenine induced the osteoblast differentiation with increased transcript levels of collage and increased ALP enzyme activity. CONCLUSIONS: Adenine exerts anti-inflammatory activity in an inflammatory cell model. Adenine benefits osteoblast differentiation in normal and inflammatory experimental settings. Adenine has a potential for the use to treat inflammatory bone condition such as osteoporosis.
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BACKGROUND: Autologous chimeric antigen receptor (CAR) T cell therapy is a promising therapeutic strategy for treating hematologic malignancies. A spectrum of serious complications caused by CAR-T cells has caught great attention. We developed a novel CAR against CD19 namely UWC19, consisting anti-CD19 single-chain variable fragment (scFv) hinged with 4-1BB and CD3z signaling domains. In this study, preclinical assessments of UWC19 were conducted to evaluate the safety and efficacy in vitro and in vivo. METHODS: To evaluate the binding activity of UWC19 cells to CD19, we measured the saturation degree of CAR with human CD19 molecules using flow cytometry in vitro. The antitumor efficacy of UWC19 cells was determined by in vitro cytotoxicity assay against CD19 positive cells and in vivo using a xenograft mouse model. Cross tissue reactivity of UWC19 cells was examined by co-culturing with cell lines from difference human tissues. Tumorigenicity was determined by subcutaneously injecting UWC19 in immunodeficient mice. Persistence was analyzed using quantitative PCR. RESULTS: We showed that UWC19 CAR T cells exerted highly specific binding affinity and cytotoxicity against CD19+ cells in vitro. In vivo, UWC19 CAR T cells are able to fully control disease progression in a Raji-xenografted immunodeficient mouse model. UWC19 exerted no obvious effects on the mean body mass and graft versus host disease were observed in surviving mice. We showed that UWC19 cells specifically recognized and eliminated CD19 positive cells, whereas CD19 negative cells were much less affected. No tumorigenicity of UWC19 in immunodeficient mice was observed. CONCLUSIONS: UWC19 treatment effectively eliminated CD19 positive tumor cells with favorable toxicity profile. The findings suggest encouraging clinical prospects for its use in patients with CD19 positive B cell malignancies. Our study presented an alternative evaluation strategy for CAR-T cell products.
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In this study we determined the in vivo activity of model ovalbumin vaccines delivered by direct intramuscular delivery of plasmid DNA or oral delivery using a recombinant suicidal Listeria monocytogenes strain (rsΔ2). In a previous report we described how rsΔ2 is capable of delivering luciferase, as protein or DNA, in vitro, into non-dividing intestinal epithelial cells (Kuo et al., 2009). This is achieved by engineering a dual expression shuttle vector, pDuLX-Luc, that replicates in E. coli and rsΔ2 and drives gene expression from the Listeria promoter (Phly) as well as the eukaryotic cytomegalovirus promoter (CMV), thereby delivering both protein and plasmid DNA to the cell cytoplasm. For the current in vivo study rsΔ2 containing pDuLX-OVA was used to deliver both ovalbumin protein and the mammalian expression plasmid by the oral route. Controls were used to investigate the activity of this system versus positive and negative controls, as well as quantifying activity against direct intramuscular injection of expression plasmids. Oral administration of rsΔ2(pDuLX-OVA) produced significant titres of antibody and was effective at inducing targeted T-cell lysis (approximately 30% lysis relative to an experimental positive control, intravenous OVA-coated splenocytes+lipopolysaccharide). Intramuscular injection of plasmids pDuLX-OVA or p3L-OVA (which lacks the prokaryotic promoter) also produced significant CTL-mediated cell lysis. The delivery of the negative control rsΔ2 (pDuLX-Luc) confirmed that the observed activity was induced specifically by the ovalbumin vaccination. The data suggest that the oral activity of rsΔ2(pDuLX-OVA) is explained by delivery of OVA protein, expressed in rsΔ2 from the prokaryotic promoter present in pDuLX-OVA, but transfection of mammalian cells in vivo may also play a role. Antibody titres were also produced by oral delivery (in rsΔ2) of the p3L-OVA plasmid in which does not include a prokaryotic promoter.
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Listeria monocytogenes/genética , Plásmidos/genética , Vacunas de ADN/genética , Vacunas de ADN/inmunología , Administración Oral , Animales , Femenino , Inmunidad Celular/genética , Inmunidad Celular/inmunología , Inmunidad Humoral/genética , Inmunidad Humoral/inmunología , Inyecciones Intramusculares , Ratones , Vacunas de ADN/administración & dosificaciónRESUMEN
House dust mite (HDM) allergens are one of the major causes leading to respiratory hypersensitiveness and airway remodeling. Here we hypothesized that a major HDM allergen Der p 2 could increase cell motility and invasiveness of non-small cell lung cancer (NSCLC) cells. Our results showed that low dose (1 and 3 µg/mL) recombinant Der p 2 protein (DP2) enhanced the migration and invasiveness of human NSCLC cell A549, H1299 and CL1-5, but nonsignificantly altered their growth. Further investigation revealed that integrin αV level was increased and its downstream signaling including focal adhesion kinase (FAK) and paxillin were activated in A549 cells exposed to DP2. In parallel, DP2 also activated the FAK-associated signaling effectors such as Src, phosphatidyl inositol 3-kinase (PI3K), AKT, p38 mitogen-activated protein kinase (P38), extracellular signal-regulated kinase 1/2 (ERK1/2) and c-Jun N-terminal kinase (JNK). Our findings also revealed that DP2 increased expression level of urokinase type plasminogen-activated kinase (uPA) and uPA receptor (uPAR), and subsequently enhanced the binding of uPAR to integrin αV. Moreover, the involvement of toll-like receptor 2/4 (TLR2/4)-triggered ERK1/2 activation in the increased expression of uPA and uPAR was also demonstrated. Collectively, these findings indicate that DP2 can enhance cell motility and invasiveness of NSCLC cells, attributing to TLR2/4-ERK1/2 activation, increased uPA and uPAR expression, enhanced binding of uPAR to integrin αV, and the consequent FAK signaling cascades. Thus, we suggest that DP2 may exacerbate NSCLC via promoting metastatic ability of carcinoma cell.
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Antígenos Dermatofagoides/farmacología , Proteínas de Artrópodos/farmacología , Carcinoma de Pulmón de Células no Pequeñas/patología , Neoplasias Pulmonares/patología , Invasividad Neoplásica/patología , Western Blotting , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Línea Celular Tumoral , Movimiento Celular/fisiología , Quinasa 1 de Adhesión Focal/metabolismo , Técnicas de Silenciamiento del Gen , Humanos , Inmunoprecipitación , Integrinas/metabolismo , Neoplasias Pulmonares/metabolismo , Reacción en Cadena de la Polimerasa , Transducción de Señal/fisiología , Receptores Toll-Like/metabolismo , Regulación hacia Arriba , Activador de Plasminógeno de Tipo Uroquinasa/biosíntesisAsunto(s)
Adenina/farmacología , Senescencia Celular/efectos de los fármacos , Dermis/citología , Folículo Piloso/efectos de los fármacos , Proteínas Quinasas Activadas por AMP/metabolismo , Aminoimidazol Carboxamida/análogos & derivados , Aminoimidazol Carboxamida/farmacología , División Celular/efectos de los fármacos , Células Cultivadas , Perfilación de la Expresión Génica , Regulación de la Expresión Génica/efectos de los fármacos , Folículo Piloso/citología , Folículo Piloso/enzimología , Humanos , NAD/metabolismo , Fosforilación/efectos de los fármacos , Procesamiento Proteico-Postraduccional/efectos de los fármacos , Ribonucleótidos/farmacología , beta-Galactosidasa/análisisRESUMEN
The AMP-activated protein kinase (AMPK) signaling system plays a key role in cellular stress by repressing the inflammatory responses induced by the nuclear factor-kappa B (NF-κB) system. Previous studies suggest that the anti-inflammatory role of AMPK involves activation by adenine, but the mechanism that allows adenine to produce these effects has not yet been elucidated. In human umbilical vein endothelial cells (HUVECs), adenine was observed to induce the phosphorylation of AMPK in both a time- and dose-dependent manner as well as its downstream target acetyl Co-A carboxylase (ACC). Adenine also attenuated NF-κB targeting of gene expression in a dose-dependent manner and decreased monocyte adhesion to HUVECs following tumor necrosis factor (TNF-α) treatment. The short hairpin RNA (shRNA) against AMPK α1 in HUVECs attenuated the adenine-induced inhibition of NF-κB activation in response to TNF-α, thereby suggesting that the anti-inflammatory role of adenine is mediated by AMPK. Following the knockdown of adenosyl phosphoribosyl transferase (APRT) in HUVECs, adenine supplementation failed to induce the phosphorylation of AMPK and ACC. Similarly, the expression of a shRNA against APRT nullified the anti-inflammatory effects of adenine in HUVECs. These results suggested that the role of adenine as an AMPK activator is related to catabolism by APRT, which increases the cellular AMP levels to activate AMPK.
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Proteínas Quinasas Activadas por AMP/metabolismo , Adenina/farmacología , Inflamación/tratamiento farmacológico , Inflamación/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Adenina/toxicidad , Adenina Fosforribosiltransferasa/genética , Adenina Fosforribosiltransferasa/metabolismo , Aminoimidazol Carboxamida/análogos & derivados , Aminoimidazol Carboxamida/farmacología , Adhesión Celular/efectos de los fármacos , Células Cultivadas , Activación Enzimática/efectos de los fármacos , Expresión Génica/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana , Humanos , Monocitos/metabolismo , FN-kappa B/metabolismo , Fosforilación/efectos de los fármacos , Transporte de Proteínas/efectos de los fármacos , ARN Interferente Pequeño/metabolismo , Ribonucleótidos/farmacologíaRESUMEN
BACKGROUND: 6-gingerol has been reported to have anti-inflammatory effects in different experimental settings. The present study aimed at evaluating the effect of 6-gingerol on dextran sodium sulfate (DSS)-induced barrier impairment and inflammation in vitro and in vivo. METHODS: a differentiated Caco-2 monolayer was exposed to DSS and treated with different concentrations of 6-gingerol (0, 1, 5, 10, 50, and 100 µM). Changes in intestinal barrier function were determined using transepithelial electrical resistance (TEER). The anti-inflammatory activity of 6-gingerol was examined as changes in the expression of proinflammatory cytokine using quantitative real-time PCR. Western blotting was employed to determine the activation of adenosine monophosphate-activated protein kinase (AMPK). Mice with DSS-induced colitis were given different oral dosages of 6-gingerol daily for 14 days. Body weight and colon inflammation were evaluated, and level of proinflammatory cytokines in colon tissues was measured. RESULTS: 6-gingerol treatment was shown to restore impaired intestinal barrier function and to suppress proinflammatory responses in DSS-treated Caco-2 monolayers. We found that AMPK was activated on 6-gingerol treatment in vitro. In animal studies, 6-gingerol significantly ameliorated DSS-induced colitis by restoration of body weight loss, reduction in intestinal bleeding, and prevention of colon length shortening. In addition, 6-gingerol suppressed DSS-elevated production of proinflammatory cytokines (IL-1ß, TNFα, and IL-12). CONCLUSION: our findings highlight the protective effects of 6-gingerol against DSS-induced colitis. We concluded that 6-gingerol exerts anti-inflammatory effects through AMPK activation. It is suggested that 6-gingerol has a promising role in treatment of IBD.
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Proteínas Quinasas Activadas por AMP/metabolismo , Catecoles/farmacología , Colitis/tratamiento farmacológico , Sulfato de Dextran/toxicidad , Alcoholes Grasos/farmacología , Proteínas Quinasas Activadas por AMP/genética , Animales , Células CACO-2 , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Colitis/inducido químicamente , Colon/efectos de los fármacos , Colon/metabolismo , Modelos Animales de Enfermedad , Femenino , Humanos , Inflamación/tratamiento farmacológico , Interleucina-12/metabolismo , Interleucina-1beta/metabolismo , Ratones , Ratones Endogámicos C57BL , Fosforilación , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Adenosine 5'-monophosphate-activated protein kinase (AMPK) is a key regulator of cellular energy homeostasis via modulating metabolism of glucose, lipid, and protein. In addition to energy modulation, AMPK has been demonstrated to associate with several important cellular events including inflammation. The results showed that ENERGI-F704 identified from bamboo shoot extract was nontoxic in concentrations up to 80 µM and dose-dependently induced phosphorylation of AMPK (Thr-172) in microglia BV2 cells. Our findings also showed that the treatment of BV2 with ENERGI-F704 ameliorated the LPS-induced elevation of IL-6 and TNF-α production. In addition, ENERGI-F704 reduced increased production of nitric oxide (NO) and prostaglandin E2 (PGE2) via downregulating the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2), respectively. Moreover, ENERGI-F704 decreased activated nuclear translocation and protein level of NF-κB. Inhibition of AMPK with compound C restored decreased NF-κB translocation by ENERGI-F704. In conclusion, ENERGI-F704 exerts inhibitory activity on LPS-induced inflammation through manipulating AMPK signaling and exhibits a potential therapeutic agent for neuroinflammatory disease.
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Proteínas Quinasas Activadas por AMP/metabolismo , Metabolismo Energético/efectos de los fármacos , Inflamación/tratamiento farmacológico , Activación Transcripcional/efectos de los fármacos , Línea Celular , Dinoprostona/biosíntesis , Metabolismo Energético/genética , Humanos , Inflamación/inducido químicamente , Inflamación/metabolismo , Lipopolisacáridos/toxicidad , Microglía/citología , Microglía/efectos de los fármacos , Óxido Nítrico/biosíntesis , Extractos Vegetales/administración & dosificación , Extractos Vegetales/química , Sasa/químicaRESUMEN
Cell migration plays a pivotal role in airway repair and remodeling involved in respiratory diseases such as asthma. Interleukin-6 (IL-6) and fascin-1 are involved in cell migration upon stimulation; however, the roles of IL-6 and fascin-1 in migration of airway epithelial cell remain sketchy. The present study was aimed to investigate influence of IL-6 on cell motility with emphasis on the association with fascin-1. Wound healing assay and transmigration assay were performed to examine effect of IL-6 on migration and invasiveness of human bronchial epithelial cell BEAS-2B. Level of mRNA expression was determined by RT-PCR and quantitative real-time RT-PCR (Q-PCR). Involvement of kinase and transcription factor signaling in IL-6-induced cell migration was investigated using immunoblot and specific inhibitors. IL-6 significantly augmented cell migration and invasiveness in parallel with elevated fascin-1 expression. Further investigation showed that IL-6 dose-dependently upregulated fascin-1 expression in both mRNA and protein levels. We showed that IL-6 activated Akt and inhibited glycogen synthase kinase-3ß (GSK-3ß), highly associating with fascin-1 mRNA expression. Additionally, IL-6-induced migration was significantly diminished by phosphatidyl inositol 3-phosphate kinase (PI3K) inhibitor (wortamannin) and ß-catenin inhibitor FH535. Moreover, LiCl and SB216763, inhibitors of GSK-3ß augmented cell migration as well as fascin-1 mRNA expression. Conclusively, these findings reveal that IL-6-induced migration of BEAS-2B cell may be attributed to activation of Akt, inhibition of GSK-3ß, and the associated increase of ß-catenin and fascin-1 expression, indicating an important role of Akt/GSK-3ß signaling and ß-catenin/fascin-1 in IL-6 associated airway remodeling.
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Proteínas Portadoras/metabolismo , Células Epiteliales/efectos de los fármacos , Glucógeno Sintasa Quinasa 3/metabolismo , Interleucina-6/farmacología , Proteínas de Microfilamentos/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Sistema Respiratorio/efectos de los fármacos , Androstadienos/farmacología , Bioensayo , Proteínas Portadoras/genética , Línea Celular Transformada , Movimiento Celular/efectos de los fármacos , Cámaras de Difusión de Cultivos , Relación Dosis-Respuesta a Droga , Células Epiteliales/citología , Células Epiteliales/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Glucógeno Sintasa Quinasa 3/antagonistas & inhibidores , Glucógeno Sintasa Quinasa 3/genética , Glucógeno Sintasa Quinasa 3 beta , Humanos , Indoles/farmacología , Cloruro de Litio/farmacología , Maleimidas/farmacología , Proteínas de Microfilamentos/genética , Fosfatidilinositol 3-Quinasas/genética , Fosfatidilinositol 3-Quinasas/metabolismo , Inhibidores de las Quinasa Fosfoinosítidos-3 , Proteínas Proto-Oncogénicas c-akt/genética , ARN Mensajero/biosíntesis , Sistema Respiratorio/citología , Sistema Respiratorio/metabolismo , Transducción de Señal/efectos de los fármacos , Sulfonamidas/farmacología , Wortmanina , Cicatrización de Heridas/efectos de los fármacos , beta Catenina/antagonistas & inhibidores , beta Catenina/genética , beta Catenina/metabolismoRESUMEN
Cystamine, a disulphide metabolite, has been demonstrated to ameliorate various lupus-associated tissue damages by animal models. However, effects of cystamine on apoptosis of cardiac tissue, a main cardiac damage attributing to lupus, are less obvious. Therefore, we aimed to investigate whether or not cystamine possesses anti-apoptotic effects with emphasis on LV tissue of lupus-prone mice NZB/W-F1. Cystamine treatment was performed by daily intraperitoneal administration. Morphology and apoptotic status of ventricular tissues in the treated mice were assessed by microscopy and TUNEL assay, respectively. Levels of apoptotic biomarkers were determined using immunoblot. Our results revealed that cystamine significantly attenuated the apoptosis of LV tissues in NZB/W-F1 mice, whereas the morphology of the tissues was slightly altered. In addition, cystamine reduced level of Fas and inhibited activation of caspase-8. Cystamine also increased level of Bcl-2 and phosphorylation of Bad, and decreased level of Bad and truncated Bid (tBid). Moreover, level of cytosolic cytochrome c and Apaf-1, and activation of caspase-9 and caspase-3 were suppressed in response to cystamine treatment. In Balb/c mice, as normal control mice, changes in cell morphology and levels of the tested apoptotic components were found insignificant in the LV tissues. These findings indicate that cystamine treatment attenuates apoptosis of LV tissues of NZB/W-F1 mice through suppressing both intrinsic and extrinsic apoptotic pathways. Therefore, cystamine is considered beneficial to alleviating lupus-associated cardiac damages.
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Apoptosis/efectos de los fármacos , Cistamina/farmacología , Ventrículos Cardíacos/patología , Lupus Eritematoso Sistémico/patología , Animales , Factor Apoptótico 1 Activador de Proteasas/metabolismo , Proteína Proapoptótica que Interacciona Mediante Dominios BH3/metabolismo , Caspasa 3/metabolismo , Caspasa 8/metabolismo , Caspasa 9/metabolismo , Citocromos c/metabolismo , Ventrículos Cardíacos/efectos de los fármacos , Ventrículos Cardíacos/metabolismo , Immunoblotting , Etiquetado Corte-Fin in Situ/métodos , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos NZB , Proteínas Proto-Oncogénicas/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2 , Proteína Letal Asociada a bcl/metabolismo , Receptor fas/metabolismoRESUMEN
BACKGROUND: Glucagon-like peptide-1 (GLP-1) and its analogues are reported to exert wide-ranging cardiovascular actions in preclinical and clinical studies. We thus investigated whether the GLP-1 receptor agonist, exendin-4, has inhibitory effects on LPS-stimulated inflammatory response in cardiomyoblasts. METHODS: H9c2 cardiomyoblasts were exposed to LPS and treated with exendin-4. Expressions of proinflammatory mediators were assessed using quantitative real-time PCR. Nuclear localization of NF-κB was examined using immunoblotting. mRNA expression of inducible nitric oxide synthase (iNOS) and nitric oxide (NO) production were evaluated by q PCR and NO assay. Furthermore, anti-apoptotic effect of exendin-4 in LPS-stimulated H9c2 cells was determined using qPCR and immunoblot. RESULTS: Exposure to LPS increased mRNA expressions of TNF-α, COX-2 and MMP-9 in H9c2 cells. It also caused increases in iNOS mRNA expression and NF-κB nuclear translocation. Exendin-4 dose-dependently downregulated mRNA levels of TNF-α, COX-2 and MMP-9 in LPS-stimulated H9c2 cells. It also reduced NF-κB nuclear translocation. Treatment with exendin-4 showed no effect on LPS-induced apoptosis in H9c2 cells. CONCLUSIONS: Exendin-4 exerts an effect on cardiomyoblast exposed to LPS by inhibiting mRNA expression of inflammatory mediators and suppressing NF-κB activation. These effects are consistent with some of the observed anti-inflammatory properties of exendin-4, as well as its beneficial actions on the cardiovascular system.
Asunto(s)
Apoptosis/efectos de los fármacos , Hipoglucemiantes/farmacología , Lipopolisacáridos/farmacología , Mioblastos Cardíacos/metabolismo , FN-kappa B/metabolismo , Péptidos/farmacología , Ponzoñas/farmacología , Transporte Activo de Núcleo Celular/efectos de los fármacos , Línea Celular , Núcleo Celular/metabolismo , Ciclooxigenasa 2/biosíntesis , Exenatida , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Metaloproteinasa 9 de la Matriz/biosíntesis , Óxido Nítrico Sintasa de Tipo II/biosíntesis , Factor de Necrosis Tumoral alfa/biosíntesisRESUMEN
Non-proteolytic group 2 allergen, Der p 2 (DP2) is known as a major allergen derived from house dust mite Dermatophagoides pteronyssinus.Paracellular epithelial barrier, being composed of a number of tight junction (TJ) molecules, plays pivotal roles in resistance of pathogen invading. However, whether DP2 affects epithelial TJ molecules is unclear. Therefore, we aimed to investigate the effects of DP2 on epithelial TJ molecules, and the mechanism by which expression of junction molecules is regulated by DP2. Cell cycle and mRNA expression of TJ proteins of lung alveolar cell A549 were analyzed by RT-PCR and flow cytometry. Level of claudin-2, subcellular distribution of b-catenin and kinase activation was determined using immunoblot. Our findings revealed that DP2 had no significant influence on cell cycle distribution but affected mRNA expression of TJ molecules including claudin-2, occludin, and ZO-1 in A549 cells. Our results showed that DP2 significantly elevated level of claudin-2 and increased expression and nuclear translocation of b-catenin. Moreover, DP2 enhanced the phosphorylation of glycogen synthase kinase-3b (GSK-3b) and its potential upstream regulator Akt. The DP2-induced claudin-2 expression was also suppressed by GSK-3b inhibitor (lithium chloride) and phosphatidyl inositol 3-phosphate kinase (PI3K) inhibitor (wortamannin). Taken together, these findings showed that DP2 increased claudin-2 expression and its cell surface distribution in A549 cells, which may attribute to phosphorylation of GSK-3b and Akt and the consequent increase and nuclear translocation of b-catenin. It is suggested that presence of DP2 may alter epithelial junction by regulating expression of TJ molecules.
Asunto(s)
Antígenos Dermatofagoides/farmacología , Glucógeno Sintasa Quinasa 3/metabolismo , Proteínas de la Membrana/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , beta Catenina/metabolismo , Animales , Proteínas de Artrópodos , Línea Celular , Claudinas , Electroforesis en Gel de Poliacrilamida , Citometría de Flujo , Glucógeno Sintasa Quinasa 3/genética , Glucógeno Sintasa Quinasa 3 beta , Humanos , Immunoblotting , Proteínas de la Membrana/genética , Ácaros , Proteínas Proto-Oncogénicas c-akt/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transducción de Señal/efectos de los fármacos , beta Catenina/genéticaRESUMEN
Ocimum gratissimum (OG) is widely used as a traditional herb for its antibacterial activity in Taiwan. Recently, antitumor effect of OG on breast cancer cell is also reported; however, the effects of OG on human pulmonary adenocarcinoma cell A549 remain unclear. Therefore, we aimed to investigate whether aqueous OG extract (OGE) affects viability of A549 cells and the signals induced by OGE in A549 cells. Cell viability assays revealed that OGE significantly and dose-dependently decreased the viability of A549 cell but not that of BEAS-2B cell. Morphological examination and DAPI staining indicated that OGE induced cell shrinkage and DNA condensation for A549 cells. Further investigation showed that OGE enhanced activation of caspase-3, caspase-9 and caspase-8 and increased protein level of Apaf-1 and Bak, but diminished the level of Bcl-2. Additionally, OGE inhibited the phosphorylation of extracellular signal-regulated kinase (ERK) yet enhanced the phosphorylation of c-Jun N-terminal kinase (JNK) and p38 MAP kinase (p38). In conclusion, our findings indicate that OGE suppressed the cell viability of A549 cells, which may result from the activation of apoptotic signaling and the inhibition of anti-apoptotic signaling, suggesting that OGE might be beneficial to lung carcinoma treatment.
RESUMEN
We have generated a recombinant stable, suicidal Listeria monocytogenes strain (rsDelta2) capable of delivering antigens as protein or DNA into nondividing intestinal epithelial cells. The rsDelta2 strain was generated by inserting a cell wall hydrolysin gene, "ply118" together with its associated holin gene from a Listeria-specific phage, into the attenuated L. monocytonegenes genome of strain Delta2. The hol118/ply118 gene was placed under the control of the Listeria promoter PactA, inducing bacteria to undergo autolysis in eukaryotic cells. The rsDelta2 strain had normal growth rate in rich bacterial growth medium, but its replication in eukaryotic cells was limited, and its autolysis was used to deliver its contents to the cytoplasm of eukaryotic cells. The delivery potential of rsDelta2 was explored using engineered shuttle vectors designed to facilitate expression of a transgene, either in rsDelta2 (driven by Phly) or in the mammalian cell (driven by P(CMV)), or both (using our engineered dual Listeria and mammalian expression vector, pDuLX). The luciferase reporter was used to demonstrate that pDuLX vector allowed delivery of both protein and DNA to dividing Caco-2 human epithelial cells. As expected, nondividing fully differentiated Caco-2 monolayers were resistant to transfection with Lipofectamine, which can be explained by lack of access to the cell nucleus. We demonstrated that when Caco-2 monolayers were treated with rsDelta2, the bacteria were able to deliver a significant quantity of luciferase protein. By implication the bacteria were also able to deliver DNA, but expression driven by the eukaryotic promoter in host Caco-2 cells was not observed. When the rsDelta2 strain was taken up by Caco-2 cells, there was little or no bacterial growth, whereas the control Delta2 strain was viable and grew by approximately three log cycles within the Caco-2 cells. A small mass of protein or DNA was delivered by the Delta2 strain perhaps because some bacteria died, but despite the level of growth the mass of protein delivered to dividing Caco-2 cells by the Delta2 strain was considerably less than that delivered by the rsDelta2 strain. We concluded that the Listeria delivery system has prospects for oral vaccination using antigens synthesized by the bacterium itself.