RESUMO
We have previously reported the protective effects of blue light-emitting diode (BLED)-stimulated cell metabolites on cell injury. To further examine the effect of conditioned media (CM) derived from BLED (5 J/cm2)-exposed human normal fibroblasts (CMBL5) for clinical application, we have used the choline chloride and phenol red-free media and then concentrated CMBL5 using a centrifugal filter unit. The collected CMBL5-lower part (CMBL5-LO) has evaluated the inflammatory protein expression profile in LPS-stimulated RAW264.7 cells. Comprehensive metabolomic profiling of CMBL5-LO was carried out using hybrid tandem mass spectrometry. Treatment with CMBL5-LO showed the cytoprotective effect on apoptotic cell death, but rather increased apoptotic cells after treatment with CMBL5-upper part (CMBL5-UP). In addition, CMBL5-LO inhibited several chemo-attractants, including interleukin (IL)-6, macrophage inflammatory protein (MIP)-2, chemokine (C-C motif) ligand 5 (CCL5), granulocyte colony-stimulating factor (GCSF), and monocyte chemoattractant protein-1 (MCP-1) expression. Pro-inflammatory nitric oxide was decreased after CMBL5-LO treatment, but not by CMBL5-UP treatment. Interestingly, treatment with CMBL5-LO stimulated expression of heme oxygenase-1, indicating its anti-inflammatory property. Most endoplasmic reticulum (ER) stress proteins except for transcription factor C/EBP homologous protein (CHOP) were highly expressed after irradiation with BLED in cells. Further studies are needed to examine the precise mechanism by CMBL5-LO in cells.
Assuntos
Anti-Inflamatórios/farmacologia , Meios de Cultivo Condicionados/farmacologia , Fibroblastos/efeitos da radiação , Luz , Animais , Apoptose/efeitos dos fármacos , Linhagem Celular , Quimiocina CXCL2/metabolismo , Cor , Citocinas/metabolismo , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Fibroblastos/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Interleucina-6/metabolismo , Lipopolissacarídeos/farmacologia , Camundongos , Óxido Nítrico/biossíntese , Substâncias Protetoras/farmacologia , Células RAW 264.7 , Regulação para Cima/efeitos dos fármacosRESUMO
PURPOSE: To investigate potential of chitosan hydrogel microparticles (CHI) for treatment of VX2 carcinoma. MATERIALS AND METHODS: Two weeks after liver VX2 implantation, contrast-enhanced computerized tomographic scanning was conducted. Rabbits (n = 2) with successful tumor growth were treated with different sizes of 99mTc-labeled CHI (60-80 µm and 100-120 µm) via intra-arterial hepatic catheterization. Liver distribution of 99mTc-labeled CHI was determined by means of autoradiography, a radiation-based photographic technique. In the next part of this study, therapeutic effectiveness was examined with the use of CHI with the size range of 60-80 µm (n = 11). Tumor growth response and levels of blood liver enzymes were studied at baseline and 1 and 2 weeks after CHI treatment. RESULTS: Successful tumor growth was confirmed in all rabbits (24/24). Intrahepatic CHI with the size range of 60-80 µm resulted in liver localization in more close proximity to tumor nodule versus 100-120 µm. Baseline tumor volume was 1,909 ± 575 mm3 in animals receiving CHI versus 1,831 ± 249 mm3 in control animals (P = .342). In control animals, tumor volume markedly increased by 1,544 ± 512% at 2 weeks after sham operation versus baseline. In animals receiving CHI, tumor volume remained relatively unchanged (54 ± 6% increase; P = .007 vs control). Levels of blood aspartate transaminase (AST) and alanine transaminase (ALT) in animals receiving CHI increased 1 week after treatment (P = .032 vs control for AST; P = .000 vs control for ALT), but returned to control levels at 2 weeks. CONCLUSIONS: CHI embolization suppressed tumor growth without appreciable damages in liver function.
Assuntos
Quitosana/farmacologia , Hidrogéis/farmacologia , Neoplasias Hepáticas Experimentais/terapia , Angiografia , Animais , Meios de Contraste , Modelos Animais de Doenças , Embolização Terapêutica , Testes de Função Hepática , Coelhos , Tomografia Computadorizada por Raios X , Carga TumoralRESUMO
This study describes the synthesis of highly water-soluble, non-toxic, and biocompatible nicotinamide adenine dinucleotide (NAD)/glucosamine (=Nga1Fh) and NAD/glucosamine/gluconic acid coated ferrihydrite nanoparticles (=Nga2Fh) and their possible uses to target tumors in living animals via 99m Tc and 125 I radioisotope labeling. The structural properties were investigated using DLS, zeta potential, TEM, FT-IR, XRD, and Raman spectroscopy. The cell toxicity in CT26 cancer cells and in vivo tumor targetability in U87MG and CT26 tumor-bearing mice was further evaluated using cRGDyK-tagged and cRGDfK-tagged ferrihydrite nanoparticles. The average diameters of the resulting Nga1Fh and Nga2Fh nanoparticles were <5 to 7 and <3 nm, respectively. The Nga2Fh nanoparticles did not show cell toxicity until 0.1 mg/mL. Using gamma camera imaging, 99m Tc-cRGDfK-Nga2Fh showed the highest tumor uptake in a U87MG tumor-bearing mouse when compared with that of 99m Tc-cRGDyK-Nga2Fh and 99m Tc-Nga2Fh. The image-based tumor-to-muscle ratio by time for 99m Tc-cRGDfK-Nga2Fh was 3.8 ± 1.7, 4.2 ± 2.0, 7 ± 1.5, 13 ± 2.0, 8 ± 3.7, and 2 ± 1.6 at 5 and 30 minutes, 1, 2, 4, and 24 hours, respectively. Although further studies are needed, the NAD/monosaccharide coated ferrihydrite nanoparticles could be presented as an interesting material for a drug delivery system.
Assuntos
Nanopartículas Metálicas/química , Neoplasias Experimentais/diagnóstico por imagem , Oligopeptídeos/química , Tomografia por Emissão de Pósitrons/métodos , Compostos Radiofarmacêuticos/síntese química , Tecnécio/química , Animais , Linhagem Celular Tumoral , Compostos Férricos/química , Glucosamina/química , Camundongos , Camundongos Nus , NAD/química , Compostos Radiofarmacêuticos/farmacocinética , Distribuição TecidualRESUMO
The aim of this study was to determine the effects and molecular mechanism of blue light emitting diode (LED) in tumor cells. A migration and invasion assay for the metastatic behavior of mouse colon cancer CT-26 and human fibrosarcoma HT-1080 cells was performed. Cancer cell migration-related proteins were identified by obtaining a 2-dimensional gel electrophoresis (2-DE) in total cellular protein profile of blue LED-irradiated cancer cells, followed by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) analysis of proteins. Protein levels were examined by immunoblotting. Irradiation with blue LED inhibited CT-26 and HT-1080 cell migration and invasion. The anti-metastatic effects of blue LED irradiation were associated with inhibition of matrix metalloproteinase (MMP)-2 and MMP-9 expression. P38 MAPK phosphorylation was increased in blue LED-irradiated CT-26 and HT-1080 cells, but was inhibited after pretreatment with SB203580, a specific inhibitor of p38 MAPK. Inhibition of p38 MAPK phosphorylation by SB203580 treatment increased number of migratory cancer cells in CT-26 and HT-1080 cells, indicating that blue LED irradiation inhibited cancer cell migration via phosphorylation of p38 MAPK. Additionally blue LED irradiation of mice injected with CT-26 cells expressing luciferase decreased early stage lung metastasis compared to untreated control mice. These results indicate that blue LED irradiation inhibits cancer cell migration and invasion in vitro and in vivo.
Assuntos
Movimento Celular/efeitos da radiação , Neoplasias do Colo/terapia , Fibrossarcoma/terapia , Luz , Fototerapia/métodos , Animais , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Neoplasias do Colo/enzimologia , Neoplasias do Colo/patologia , Eletroforese em Gel Bidimensional , Feminino , Fibrossarcoma/enzimologia , Fibrossarcoma/patologia , Humanos , Neoplasias Pulmonares/enzimologia , Neoplasias Pulmonares/prevenção & controle , Neoplasias Pulmonares/secundário , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Invasividade Neoplásica , Fosforilação , Inibidores de Proteínas Quinases/farmacologia , Proteômica/métodos , Transdução de Sinais/efeitos da radiação , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Fatores de Tempo , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
PURPOSE: To determine whether chitosan hydrogel nanoparticles loaded with vascular endothelial growth factor (VEGF) peptides (81-91 fragments) capable of targeting the ischemic myocardium enhance angiogenesis and promote therapeutic effects and whether radionuclide image-guided dosage control is feasible. MATERIALS AND METHODS: Experimental procedures and protocols were approved by the Institutional Animal Care and Use Committee. Rats (n = 32, eight per group) were subjected to myocardial ischemia (control group) and received chitosan hydrogel nanoparticles with VEGF165 proteins (chitosan VEGF) or VEGF81-91 peptides (chitosan peptides) via apical puncture. Ischemic hearts receiving chitosan without angiogenic factors served as the chitosan control. Myocardial perfusion was examined 7 days after surgery by using technetium 99m ((99m)Tc) tetrofosmin (37 MBq) autoradiography, and changes in vascular density with immunohistochemical staining were reviewed. Kruskal-Wallis test and Bonferroni corrected Mann-Whitney U test were used for multiple comparisons. Wilcoxon signed rank test was used to compare myocardial retention of (99m)Tc chitosan. RESULTS: Thirty minutes of myocardial ischemia resulted in perfusion defects (median, 54%; interquartile range [IQR], 41%-62%). Chitosan VEGF decreased perfusion defect extent (median, 68%; IQR, 63%-73%; P = .006 vs control) and increased vascular density (median, 81 vessels per high-power field; IQR, 72-100; P = .009 vs control). Administration of chitosan peptides reduced the degree of perfusion defects (median, 66%; IQR, 62%-73%; P = .006 vs control) and increased vascular density (median, 82 vessels; IQR, 78-92; P = .006 vs control). The effects of chitosan peptides on perfusion and vascular density were comparable to those seen with chitosan VEGF proteins (P = .713 and P = .833, respectively). Chitosan radiolabeled with (99m)Tc was administered twice at reperfusion with a 1-hour interval to determine whether image-guided dosage control is feasible. The hearts initially retained 4.6% (IQR, 4.1%-5.0%) of (99m)Tc chitosan administered and 9.2% (IQR, 6.6%-12.7%; P = .068) with subsequent injection. CONCLUSION: VEGF peptides have angiogenic potential and resulted in therapeutic effectiveness. Adjunct use of single photon emission computed tomography was also demonstrated for individualized treatment of myocardial ischemia by further tailoring the therapeutic dosing. Online supplemental material is available for this article.
Assuntos
Isquemia Miocárdica/diagnóstico por imagem , Isquemia Miocárdica/tratamento farmacológico , Nanopartículas , Tomografia Computadorizada de Emissão de Fóton Único , Fator A de Crescimento do Endotélio Vascular/farmacologia , Animais , Autorradiografia , Quitosana/farmacologia , Hidrogéis/farmacologia , Imuno-Histoquímica , Masculino , Imagem Molecular/métodos , Reperfusão Miocárdica , Compostos Organofosforados , Compostos de Organotecnécio , Compostos Radiofarmacêuticos , Ratos , Ratos Sprague-Dawley , Tomografia Computadorizada por Raios XRESUMO
This experimental study aimed to evaluate the antiviral and synergistic effects of photoenergy irradiation on human herpes simplex virus type I (HSV-1) infection. We assessed viral replication, plaque formation, and relevant viral gene expression to examine the antiviral and synergistic effects of blue light (BL) with acyclovir treatment. Our results showed that daily BL (10 J/cm2) irradiation inhibited plaque-forming ability and decreased viral copy numbers in HSV-1-infected monkey kidney epithelial Vero cells and primary human oral keratinocyte (HOK) cells. Combined treatment with the antiviral agent acyclovir and BL irradiation increased anti-viral activity, reducing viral titers and copy numbers. In particular, accumulated BL irradiation suppressed characteristic viral genes including UL19 and US6, and viral DNA replication-essential genes including UL9, UL30, UL42, and UL52 in HOK cells. Our results suggest that BL irradiation has anti-viral and synergistic properties, making it a promising therapeutic candidate for suppressing viral infections in clinical trials.
Assuntos
Aciclovir , Antivirais , Herpesvirus Humano 1 , Replicação Viral , Antivirais/farmacologia , Animais , Herpesvirus Humano 1/efeitos dos fármacos , Herpesvirus Humano 1/efeitos da radiação , Herpesvirus Humano 1/fisiologia , Herpesvirus Humano 1/genética , Chlorocebus aethiops , Células Vero , Humanos , Replicação Viral/efeitos dos fármacos , Replicação Viral/efeitos da radiação , Aciclovir/farmacologia , Luz , Herpes Simples/virologia , Herpes Simples/tratamento farmacológico , Queratinócitos/virologia , Queratinócitos/efeitos da radiação , Queratinócitos/efeitos dos fármacos , Ensaio de Placa ViralRESUMO
The aim of this study was to investigate the antiviral and anti-inflammatory functions of blue light (BL) in cutaneous viral infections. Previously, we examined the photo-biogoverning role of 450 nm BL in SARS-CoV-2-infected cells, which showed that photo-energy could inhibit viral activation depending on the number of photons. However, the communication network between photo-energy irradiation and immune cells involved in viral infections has not been clarified. We verified viral activation, inflammatory responses, and relevant downstream cascades caused by human simplex virus type I (HSV-1) after BL irradiation. To examine the antiviral effect of BL, we further tested whether BL could disturb viral absorption or entry into host cells. The results showed that BL irradiation, but not green light (GL) exposure, specifically decreased plaque-forming activity and viral copy numbers in HSV-1-infected cells. Accumulated BL irradiation inhibited the localization of viral proteins and the RNA expression of characteristic viral genes such as UL19, UL27, and US6, thus exerting to an anti-viral effect. The results also showed that BL exposure during viral absorption interfered with viral entry or destroyed the virus, as assessed by plaque formation and quantitative PCR assays. The levels of the pro-inflammatory mediators interleukin (IL)-18 and IL-1ß in M1-polarized macrophages were increased by HSV-1 infection. However, these increases were attenuated by BL irradiation. Importantly, BL irradiation decreased cGAS and STING expression, as well as downstream NF-κB p65, in M1-polarized HSV-1-infected macrophages, demonstrating anti-viral and anti-inflammatory properties. These findings suggest that BL could serve as an anti-viral and anti-inflammatory therapeutic candidate to treat HSV-1 infections.
Assuntos
COVID-19 , Herpesvirus Humano 1 , Humanos , Antivirais/farmacologia , Herpesvirus Humano 1/genética , Replicação Viral , SARS-CoV-2 , Anti-Inflamatórios/farmacologiaRESUMO
The aim of this study was to examine the inhibitory effect of blue light (BL) on the proliferation of metastatic cancer cells and synergistic properties with chemo-drugs. BL significantly inhibited the proliferation of B cell lymphoma (A20 and RAMOS) cells in a dose-dependent manner. Anti-proliferative effect of BL irradiation was identified to be associated with the inhibition of proliferating-cell nuclear antigen expression and cell cycle by decreasing S-phase cells. Consistent with its inhibitory effects, BL irradiation at 20 J/cm2 daily for 10 days inhibited metastasis of cancer cells which were distributed and invaded to other organs including bone marrow, liver, kidney, etc., and induced paraplegia, thereby leading to an increased survival rate of tumor-bearing mice. Anti-proliferative activity of BL was expanded in solid tumor cells including pancreatic carcinoma (Mia PaCa-2, PANC-1), lung carcinoma A549 and colorectal carcinoma HCT116 cells. Additionally, combination with chemo-drugs such as 5-FU and gemcitabine resulted in an increase in the anti-proliferative activity after BL irradiation accompanied by regulating mRNA translational process via inhibition of p70S6K, 4EBP-1 and eIF4E phosphorylation during cellular proliferation. These results indicate the anti-metastatic and photo-biogoverning abilities of BL irradiation as a potent therapeutic potential for repressing the progression of tumor cells.
Assuntos
Antineoplásicos , Neoplasias , Animais , Camundongos , Apoptose , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Processamento de Proteína Pós-Traducional , Gencitabina , Proliferação de Células , Linhagem Celular TumoralRESUMO
Although microRNA-21 (miR-21) is emerging as an oncogene and has been shown to target several tumor suppressor genes, including programmed cell death 4 (PDCD4), its precise mechanism of action on cancer stem cells (CSCs) is unclear. Herein, we report that FOLFOX-resistant HCT-116 and HT-29 cells that are enriched in CSCs show a 3- to 7-fold upregulation of pre- and mature miR-21 and downregulation of PDCD4. Likewise, overexpression of miR-21 in HCT-116 cells, achieved through stable transfection, led to the downregulation of PDCD4 and transforming growth factor beta receptor 2 (TGFßR2). In contrast, the levels of ß-catenin, TCF/LEF activity and the expression of c-Myc, Cyclin-D, which are increased in CSCs, are also augmented in miR-21 overexpressing colon cancer cells, accompanied by an increased sphere forming ability in vitro and tumor formation in SCID mice. Downregulation of TGFßR2 could be attributed to decreased expression of the receptor as evidenced by reduction in the activity of the luciferase gene construct comprising TGFßR2-3' untranslated region (UTR) sequence that binds to miR-21. Moreover, we observed that downregulation of miR-21 enhances luciferase-TGFßR2-3' UTR activity suggesting TGFßR2 as being one of the direct targets of miR-21. Further support is provided by the observation that transfection of TGFßR2 in HCT-116 cells attenuates TCF/LEF luciferase activity, accompanied by decreased expression of ß-catenin, c-Myc and Cyclin-D1. Our current data suggest that miR-21 plays an important role in regulating stemness by modulating TGFßR2 signaling in colon cancer cells.
Assuntos
Neoplasias do Colo/patologia , MicroRNAs/fisiologia , Células-Tronco Neoplásicas/patologia , Proteínas Serina-Treonina Quinases/fisiologia , Receptores de Fatores de Crescimento Transformadores beta/fisiologia , Proteínas Reguladoras de Apoptose/fisiologia , Neoplasias do Colo/tratamento farmacológico , Regulação para Baixo , Resistencia a Medicamentos Antineoplásicos , Células HCT116 , Células HT29 , Humanos , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Proteínas de Ligação a RNA/fisiologia , Receptor do Fator de Crescimento Transformador beta Tipo II , Receptores de Fatores de Crescimento Transformadores beta/antagonistas & inibidores , Transdução de Sinais , Via de Sinalização Wnt , beta Catenina/fisiologiaRESUMO
Most recently, severe acute respiratory syndrome coronavirus-2 has triggered a global pandemic without successful therapeutics. The goal of the present study was to define the antiviral effect and therapeutic action of blue light irradiation in SARS-CoV-2-infected cells. Vero cells were infected with SARS-CoV-2 (NCCP43326) or mock inoculum at 50 pfu/well. After blue light irradiation, the inhibitory effect was assessed by qPCR and plaque reduction assay. When Vero cells were irradiated to blue light ranging from 1.6 to 10 J cm-2 , SARS-CoV-2 replication was inhibited by up to 80%. The antiviral effect of blue light irradiation was associated with translation suppression via the phosphorylation of eIF2α by prolonging endoplasmic reticulum (ER) stress. The levels of LC3A/B and Beclin-1, which are key markers of autophagy, and the levels of PERK and PDI for ER stress were highly increased, whereas caspase-3 cleavage was inhibited after blue light irradiation in the later stage of infection. Our data revealed that blue light irradiation exerted antiviral and photo-biogoverning activities by prolonging ER stress and stimulating autophagy progression during viral infection. The findings increase our understanding of how photo-energy acts on viral progression and have implications for use in therapeutic strategies against COVID-19.
Assuntos
COVID-19 , Animais , COVID-19/radioterapia , Chlorocebus aethiops , Pandemias , SARS-CoV-2 , Células Vero , Replicação ViralRESUMO
We have previously demonstrated that expression of the novel gene schlafen-3 (Slfn-3) correlates with intestinal epithelial cell differentiation (Patel VB, Yu Y, Das JK, Patel BB, Majumdar AP. Biochem Biophys Res Commun 388: 752-756, 2009). The present investigation was undertaken to examine whether Slfn-3 plays a role in regulating differentiation of FOLFOX-resistant (5-fluorouracil + oxaliplatin) colon cancer cells that are highly enriched in cancer stem cells (CSCs). Transfection of Slfn-3 in FOLFOX-resistant colon cancer HCT-116 cells resulted in increase of alkaline phosphatase activity, a marker of intestinal differentiation. Additionally, Slfn-3 transfection resulted in reduction of mRNA and protein levels of the CSC markers CD44, CD133, CD166, and aldehyde dehydrogenase 1 in both FOLFOX-resistant HCT-116 and HT-29 cells. This was accompanied by decreased formation of tumorosphere/colonosphere (an in vitro model of tumor growth) in stem cell medium and inhibition of expression of the chemotherapeutic drug transporter protein ABCG2. Additionally, Slfn-3 transfection of FOLFOX-resistant HCT-116 and HT-29 cells reduced Hoechst 33342 dye exclusion. Finally, Slfn-3 transfection inhibited the expression of transforming growth factor-α in both FOLFOX-resistant colon cancer cells, but stimulated apoptosis in response to additional FOLFOX treatment. In summary, our data demonstrate that Slfn-3 expression inhibits multiple characteristics of CSC-enriched, FOLFOX-resistant colon cancer cells, including induction of differentiation and reduction in tumorosphere/colonosphere formation, drug transporter activity, and autocrine stimulation of proliferation. Thus Slfn-3 expression may render colon CSCs more susceptible to cancer chemotherapeutics.
Assuntos
Comunicação Autócrina/genética , Proteínas de Ciclo Celular/genética , Neoplasias do Colo/tratamento farmacológico , Resistencia a Medicamentos Antineoplásicos/genética , Células-Tronco Neoplásicas/metabolismo , Proteínas/genética , Proteínas/fisiologia , RNA Mensageiro/metabolismo , Antígeno AC133 , Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP , Transportadores de Cassetes de Ligação de ATP/metabolismo , Família Aldeído Desidrogenase 1 , Antígenos CD/metabolismo , Protocolos de Quimioterapia Combinada Antineoplásica , Apoptose , Moléculas de Adesão Celular Neuronais/metabolismo , Proteínas de Ciclo Celular/fisiologia , Diferenciação Celular/genética , Neoplasias do Colo/genética , Receptores ErbB/metabolismo , Proteínas Fetais/metabolismo , Fluoruracila , Glicoproteínas/metabolismo , Células HCT116 , Células HT29 , Humanos , Receptores de Hialuronatos/metabolismo , Isoenzimas/metabolismo , Leucovorina , Proteínas de Neoplasias/metabolismo , Compostos Organoplatínicos , Peptídeos/metabolismo , Retinal Desidrogenase/metabolismo , Transdução de Sinais/genética , Transfecção , Fator de Crescimento Transformador alfa/metabolismoRESUMO
The aim of the present study was to evaluate inhibitory effect of CTB glycoprotein isolated from Cudrania tricuspidata Bureau on DEHP-induced cell proliferation in lymphocytes. Our results revealed that DEHP increased lymphocyte proliferation as confirmed by increasing [(3)H]thymidine incorporation, and proliferating cell nuclear antigen (PCNA), cyclin D1, and cyclin-dependent kinase (CDK)-4 expression. This was accompanied by induced intracellular Ca(2+) level, protein kinase C (PKC) translocation from cytosol to membrane, ERK1/2 phosphorylation, and nuclear factor (NF)-κB transcriptional activation in DEHP-treated cells. However, CTB glycoprotein (100 µg/ml) reduced labeled thymidine incorporation and PCNA expression in DEHP-treated cells. Additionally CTB glycoprotein reduced Ca(2+) level, PKC translocation, ERK1/2 phosphorylation, NF-κB transcriptional activation, cell cycle proteins (cyclin D1 and CDK4) expression in cells. The activation of NF-κB was collectively blocked by pretreatment with PKC inhibitor (staurosporine) and ERK1/2 inhibitor (PD98059), respectively. The results from these experiments indicate that CTB glycoprotein inhibits cell proliferation via down regulations of Ca(2+)/PKC, ERK1/2, and cell cycle proteins induced by DEHP. Therefore, we suggest that the CTB glycoprotein might be one component for prevention of cell proliferation-related immune diseases.
Assuntos
Linfócitos B/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Dietilexilftalato/farmacologia , Glicoproteínas/farmacologia , Moraceae/química , Transdução de Sinais/efeitos dos fármacos , Animais , Linfócitos B/citologia , Linfócitos B/fisiologia , Cálcio/metabolismo , Proteínas de Ciclo Celular/metabolismo , Células Cultivadas , Regulação para Baixo , Glicoproteínas/metabolismo , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Proteína Quinase C/metabolismo , Baço/citologiaRESUMO
The aim of this study was to evaluate inhibitory effect of glycoprotein isolated from Cudrania tricuspidata Bureau (CTB glycoprotein) on allergic responses. We evaluated the activities of mitogen-activated protein kinase (MAPK), transcriptional factor, and production of immunoglobulin (Ig)E and interleukin (IL)-4 in RBL-2H3 cells and BALB/c mice. Our results showed that CTB glycoprotein inhibited the production of IgE and IL-4 in serum from ovalbumin (OVA)-treated BALB/c mice. We also found that CTB glycoprotein inhibited the phosphorylation of p38 MAPK, and transcriptional activation of nuclear factor (NF)-κB in RBL-2H3 cells. The activation of NF-κB was effectively blocked by treatment with p38 MAPK inhibitor (SKF86002). The results from these experiments indicate that the CTB glycoprotein inhibits release of ß-hexosaminidase, and production of IgE and IL-4 via down regulation of MAPK/ NF-κB on the stage of mast cell degranulation. In conclusion, we suggest that the CTB glycoprotein might be a potent preventive agent in allergic responses.
Assuntos
Glicoproteínas/farmacologia , Imunoglobulina E/sangue , Interleucina-4/sangue , Moraceae/química , NF-kappa B/metabolismo , Proteínas de Plantas/farmacologia , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Animais , Anti-Inflamatórios/isolamento & purificação , Anti-Inflamatórios/farmacologia , Degranulação Celular/efeitos dos fármacos , Linhagem Celular , Ensaio de Imunoadsorção Enzimática , Glicoproteínas/isolamento & purificação , Glicoproteínas/metabolismo , Hexosaminidases/metabolismo , Immunoblotting , Interleucina-4/genética , Masculino , Mastócitos/efeitos dos fármacos , Mastócitos/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/imunologia , Fosforilação/efeitos dos fármacos , Proteínas de Plantas/isolamento & purificação , Transdução de Sinais/efeitos dos fármacosRESUMO
Phthalate esters as plasticizers have been widespread in the environment and may be associated with development of allergic diseases such as asthma and atopic dermatitis. In this study, we demonstrated that the CTB glycoprotein attenuates allergic reactions caused by di(2-ethylhexyl) phthalate (DEHP) in human mast cells (HMC-1). This experiment evaluated degranulation of histamine and ß-hexosaminidase as well as activities of protein kinase C (PKC), stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK), activator protein (AP)-1 and interleukin (IL)-4 and tumor necrosis factor (TNF)-α using immunoblotting and reverse transcription-polymerase chain reaction (RT-PCR). Our results revealed that the CTB glycoprotein in the presence of DEHP inhibits degranulation of mast cell, translocation of PKC from cytosol to membrane, and phosphorylation of SAPK/JNK in HMC -1 cells. We also found that the CTB glycoprotein (100 µg mL(-1) ) has suppressive effects on transcriptional activation of AP-1, and on the expression of IL-4 and TNF-α in DEHP-treated HMC-1 cells. We suggest that the CTB glycoprotein inhibits degranulation of mast cells and expressions of cytokines in HMC-1 cells.
Assuntos
Antialérgicos/farmacologia , Dietilexilftalato/toxicidade , Estrogênios/toxicidade , Glicoproteínas/farmacologia , Hipersensibilidade/metabolismo , Proteínas de Plantas/farmacologia , Linhagem Celular , Histamina/metabolismo , Humanos , Hipersensibilidade/tratamento farmacológico , Interleucina-4/antagonistas & inibidores , Interleucina-4/genética , Interleucina-4/metabolismo , Proteínas Quinases JNK Ativadas por Mitógeno/antagonistas & inibidores , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Mastócitos , Moraceae/metabolismo , Fosforilação/efeitos dos fármacos , Extratos Vegetais/farmacologia , Proteína Quinase C/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator de Transcrição AP-1/metabolismo , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , beta-N-Acetil-Hexosaminidases/antagonistas & inibidores , beta-N-Acetil-Hexosaminidases/metabolismoRESUMO
BACKGROUND: Pharmacological and non-pharmacological therapies have been used to treat patients with chemotherapy-induced peripheral neuropathy (CIPN). However, the effect of therapies in cancer patients has yet to be investigated comprehensively. We hypothesized that cyclic thermal therapy would improve blood flow and microcirculation and improve the symptoms driven by CIPN. METHODS: The criteria of assessment were blood volume in region of interest (ROI) in the images, and European Organization for Research and Treatment of Cancer-Quality of Life Questionnaire-Chemotherapy-Induced Peripheral Neuropathy 20 questionnaire scores. The blood volume was quantified by using red blood cell (RBC) scintigraphy. All patients were treated 10 times during 10 days. The thermal stimulations, between 15° and 41°, were repeatedly delivered to the patient's hands. RESULTS: The total score of the questionnaires, the score of questions related to the upper limbs, the score of questions closely related to the upper limbs, and the score excluding the upper limbs questions was decreased. The blood volume was decreased, and the variance of blood volume was decreased. During cooling stimulation, the blood volume was decreased, and its variance was decreased. During warming stimulation, the blood volume was decreased, and its variance was decreased. CONCLUSIONS: We suggest that cyclic thermal therapy is useful to alleviate CIPN symptoms by blood circulation improvement. RBC scintigraphy can provide the quantitative information on blood volume under certain conditions such as stress, as well as rest, in peripheral tissue.
RESUMO
This study investigated the inhibitory effect of a glycoprotein isolated from Cudrania tricuspidata Bureau (CTB glycoprotein) on di(2-ethylhexyl) phthalate (DEHP)-induced mast cell degranulation and related signaling cascade in RBL-2H3 cells. This experiment evaluated the intracellular Ca(2+) level, and the activities of protein kinase C (PKC), mitogen-activated protein kinase (MAPK), transcription factor, and the cytokines in DEHP-treated RBL-2H3 cells. Our results revealed that the CTB glycoprotein in the presence of DEHP inhibits the release of histamine and expression of interleukin (IL)-4, IL-6, and TNF-alpha in RBL-2H3 cells. We also found that the CTB glycoprotein inhibits the intracellular Ca(2+) level, translocation of PKC from cytosol to membrane and the phosphorylation of ERK1/2 in cells. Moreover, the CTB glycoprotein (100 microg/ml) has suppressive effects on transcriptional activation of nuclear factor (NF)-kappaB in DEHP-treated RBL-2H3 cells. The activation of NF-kappaB was collectively blocked by treatment with PKC inhibitor (staurosporine) as well as ERK1/2 inhibitor (PD98059), respectively. The results from these experiments indicated that the CTB glycoprotein inhibits release of histamine and expressions of IL-4, IL-6, and TNF-alpha via down regulations of PKC/MAPK and NF-kappaB on the stage of mast cell degranulation induced by DEHP. Moreover, oral administration of CTB glycoprotein (10-20 mg/kg) inhibited compound 48/80-mediated systemic reaction in mice. In conclusion, we speculated that the CTB glycoprotein might be one component for preparation of health supplements for prevention of allergic immune disorders.
Assuntos
Antialérgicos/farmacologia , Glicoproteínas/farmacologia , Hipersensibilidade/metabolismo , Inflamação/metabolismo , Mastócitos/efeitos dos fármacos , Moraceae/química , Anafilaxia/induzido quimicamente , Anafilaxia/prevenção & controle , Animais , Western Blotting , Degranulação Celular/efeitos dos fármacos , Citocinas/efeitos dos fármacos , Citocinas/metabolismo , Dietilexilftalato/farmacologia , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Expressão Gênica/efeitos dos fármacos , Hipersensibilidade/tratamento farmacológico , Hipersensibilidade/imunologia , Inflamação/tratamento farmacológico , Inflamação/imunologia , Mastócitos/metabolismo , Moraceae/imunologia , Fosforilação , Proteína Quinase C/metabolismo , Transporte Proteico/efeitos dos fármacos , Ratos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/imunologia , p-Metoxi-N-metilfenetilamina/toxicidadeRESUMO
The purpose of this study is to investigate the inhibitory effect of a glycoprotein (CTB glycoprotein, 75 kDa) isolated from Cudrania tricuspidata Bureau (CTB) on the di-(2-ethylhexyl)phthalate (DEHP) induced expression of allergic-inflammation-related mediators in human mast cells. The changes on the levels of intracellular reactive oxygen species (ROS), mitogen-activated protein kinase (MAPK), transcription factor [nuclear factor (NF)-kappaB], and allergic inflammatory mediators [cyclooxygenase (COX)-2 and interleukin (IL)-1beta] were evaluated using Western blot and RT-PCR. Our results showed that the CTB glycoprotein in the presence of DEHP inhibits the production of intracellular ROS, the phosphorylation of ERK1/2, and p38 MAPK in HMC-1 cells. In addition, the CTB glycoprotein has suppressive effects on the transcriptional activation of NF-kappaB, and on the expression levels of COX-2 and IL-1beta in DEHP-treated HMC-1 cells. In conclusion, the CTB glycoprotein has a strong anti-inflammatory effect on the activities of allergic inflammatory mediators indirectly caused by DEHP in HMC-1 cells.
Assuntos
Dietilexilftalato/farmacologia , Glicoproteínas/farmacologia , Interleucina-1beta/metabolismo , Mastócitos/imunologia , Proteínas de Plantas/farmacologia , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Humanos , Interleucina-1beta/genética , Mastócitos/efeitos dos fármacos , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Moraceae/química , NF-kappa B/genética , NF-kappa B/metabolismo , Espécies Reativas de Oxigênio/metabolismoRESUMO
The aim of the present study was to evaluate immunomodulatory and hepatoprotective effects of glycoprotein isolated from Morus indica Linne (MIL glycoprotein) on carbon tetrachloride (CCl(4) )-induced liver injury. In the present study, MIL glycoprotein (5 and 10 mg kg(-1) body weight) was administered to ICR mice for 7 days prior to CCl(4) treatment. We evaluated the activities of alanine aminotransferase (ALT), lactate dehydrogenase (LDH), and thiobarbituric acid-reactive substances (TBARS), and expression of inflammation-related mediators including cyclooxygenase (COX)-2, tumor necrosis factor (TNF)-α, and interleukin (IL)-1 beta in CCl(4) -treated mice. Our results revealed that MIL glycoprotein reduced the activities of ALT, LDH and TBARS in serum from CCl(4) -treated mice. We also found that MIL glycoprotein reduced the activity of COX-2 and expression of TNF-α and IL-1 beta in liver from CCl(4) -treated mice. Moreover, administration of MIL glycoprotein suppressed on stress-activated protein kinase/c-jun N-terminal kinase phosphorylation and activator protein-1 transcriptional activation in livers from CCl(4) -treated mice. The results from these experiments indicate that MIL glycoprotein effectively protects against liver injury, mainly through downregulation of oxidative stress and the inflammatory response. In conclusion, we suggest that the MIL glycoprotein might be one component of health supplements for prevention of liver diseases.
Assuntos
Tetracloreto de Carbono/toxicidade , Doença Hepática Induzida por Substâncias e Drogas/prevenção & controle , Glicoproteínas/farmacologia , Mediadores da Inflamação/metabolismo , Alanina Transaminase/metabolismo , Animais , Intoxicação por Tetracloreto de Carbono/metabolismo , Intoxicação por Tetracloreto de Carbono/prevenção & controle , Ciclo-Oxigenase 2/metabolismo , Avaliação Pré-Clínica de Medicamentos , Glicoproteínas/metabolismo , Interleucina-1beta/metabolismo , Fígado/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos ICR , Morus/química , Estresse Oxidativo , Proteínas de Plantas/farmacologia , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Unlike normal cells, cancer cells mutate to thrive in exaggerated levels of reactive oxygen species (ROS). This potentially makes them more susceptible to small molecule-induced oxidative stress. The intracellular ROS increase in cancer cells is a potential area under investigation for the development of cancer therapeutics targeting cancer cells. Visible photons of 430-490 nm wavelengths from a blue-light emitting diode (BLED) encompass the visible region of the spectrum known to induce ROS in cancer cells. Curcuminoids (CUR) naturally occurring photosensitizers sensitized by the blue wavelength of the visible light, well known for its potent anti-inflammatory and anticancer activity. Poor solubility and bioavailability, of the compound of the small molecule CUR restrict the therapeutic potential and limits CUR to be used as a photosensitizer. Here, our research group reports the use of small molecules CUR, encapsulated in liposome nanocarriers (LIP-CUR) coupled with blue light-emitting diode (BLED) induced photodynamic therapy (BLED-PDT). In A549 cancer cells in vitro, LIP-CUR coupled with BLED initiated BLED-PDT and triggered 1O2, ultimately resulting in caspase-3 activated apoptotic cell death. The combination of a non-cytotoxic dose of small molecule CUR co-treated with BLED to trigger BLED-PDT could be translated and be developed as a novel strategy for the treatment of cancer.
Assuntos
Diarileptanoides/administração & dosagem , Nanopartículas/administração & dosagem , Fotoquimioterapia , Fármacos Fotossensibilizantes/administração & dosagem , Células A549 , Apoptose/efeitos dos fármacos , Humanos , Luz , Lipossomos , Neoplasias/tratamento farmacológicoRESUMO
PURPOSE: The objective of this study was to describe to develop methods of rodent leukocyte isolation and radiolabeling for in vivo inflammation imaging. METHODS: Thigh muscle inflammation was induced by injection of collagenase. Blood was collected from the jugular vein and separated by Histopaque. The collected cells were incubated in a 37 °C CO2 incubator for 1~2 h. After incubation, 99mTc-HMPAO and 18F-FDG were used to treat leukocytes followed by incubation for 30 min. 99mTc-HMPAO and 18F-FDG labeled autologous leukocytes were injected into the tail veins of rats. The images were then acquired at various time points. Image-based lesion to normal muscle ratio was compared. RESULTS: After Histopaque separation, the proportion of lymphocytes was higher than that of other cell types. After CO2 incubation, the collected leukocytes were viable, while room temperature exposed leukocytes without CO2 incubation were non-viable. Granulocytes, especially, were more quickly influenced by various conditions than the mononuclear cells. Labeling efficiencies of 99mTc-HMPAO and 18F-FDG were 4.00 ± 2.06 and 1.8%, respectively. 99mTc-HMPAO- and 18F-FDG-labeled leukocytes targeted well the inflamed lesion. 99mTc-HMPAO-labeled leukocytes, but not 18F-FDG-labeled leukocytes, were found in the abdomen activity. CONCLUSION: Inflamed lesions of rats were well visualized using autologous radiolabeled leukocytes. This method might provide good information for understanding inflammatory diseases.