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1.
Clinics ; 76: e1713, 2021. tab, graf
Article in English | LILACS | ID: biblio-1153987

ABSTRACT

OBJECTIVES: The chemokine ligand (CCL) 21 regulates the maturation, migration, and function of dendritic cells, and has been implicated in the pathogenesis of asthma. This study aimed to investigate the association between serum CCL21 levels and asthma control. METHODS: The serum levels of CCL21 and other inflammatory cytokines were analyzed in patients with asthma (n=44) and healthy controls (n=35) by enzyme-linked immunosorbent assay. IgE levels and eosinophil counts were determined by turbidimetric inhibition immunoassay and fully automatic blood analysis, respectively. The Asthma Control Test (ACT) questionnaire was used, and spirometry and fractional exhaled nitric oxide (FENO) measurements were performed. A multiple unpaired Student's t-test was performed to analyze the differences in CCL21 and interleukin levels between patients with asthma and healthy controls. The correlation of CCL21 levels with disease severity was evaluated using the Pearson's rank correlation test. RESULTS: Serum CCL21 levels were lower in patients with asthma (254.78±95.66 pg/mL) than in healthy controls (382.95±87.77 pg/mL) (p<0.001). Patients with asthma had significantly higher levels of IL-1β (19.74±16.77 vs. 2.63±5.22 pg/mL), IL-6 (7.55±8.65 vs. 2.37±2.47 pg/mL), and tumor necrosis factor-α (12.70±12.03 vs. 4.82±3.97 pg/mL) compared with the controls. CCL21 levels were positively correlated with the ACT score (rs=0.1653, p=0.0062), forced expiratory volume in 1s (FEV1)/forced vital capacity (rs=0.3607, p<0.0001), and FEV1 (rs=0.2753, p=0.0003), and negatively correlated with FENO (rs=0.1060, p=0.0310). CCL21 levels were negatively correlated with serum IgE levels (rs=0.1114, p=0.0268) and eosinophil counts (rs=0.3476, p<0.0001). CONCLUSIONS: Serum CCL21 levels may be a new biomarker for assessing asthma control.


Subject(s)
Humans , Adult , Asthma , Chemokine CCL21/blood , Forced Expiratory Volume , Chemokines , Exhalation , Ligands , Nitric Oxide
2.
Article in Spanish | LILACS, COLNAL | ID: biblio-1292441

ABSTRACT

Introducción: la malaria es uno de los mayores retos de la salud pública mundial. Es causada principalmente por los parásitos Plasmodium falciparum y Plasmodium vivax. Durante el proceso de invasión, se encuentran involucra-das las proteínas homólogas de unión a reticulocitos de P.falciparumPfRH1, PfRH2a, PfRH2b, PfRH4 y PfRH5, que tras su unión a receptores específicos de membrana permiten la invasión del merozoíto al eritrocito. Objetivo: compilar y resumir las características moleculares y estructurales de las interacciones entre las proteínas pertenecientes a la familia de proteínas homólogas de unión a reticulocitos de P.falciparum y los receptores expre-sados en la célula del hospedero. Método: revisión descriptiva sobre las proteínas homólogas de unión a reticulocitos de P. falciparum involucradas en el proceso de invasión al eritrocito. Esta revisiónincluye literatura publicada hasta el año 2020 en bases de datos electrónicas especializadas en investigación biomédica. Se encontraron 105 documentos, de los cuales se se-leccionaron 70 y se excluyeron 11, por no presentar los criterios de inclusión, analizando un total de 59 referencias. Conclusión: la invasión del merozoíto es mediada por interacciones específicas de los ligandos de las familias EBL y PfRH. La unión de las proteínas PfRH1 y PfRH2b a sus receptores en el eritrocito da lugar a la liberación de la proteína EBL-175 que, junto con PfRH4, median la formación de una unión estrecha entre el parásito y los glóbulos rojos. Ello permite la unión de la proteína PfRH5 a la basigina y la entrada del parásito a la célula del hospedero


Introduction: Malaria is one of the world's greatest public health challenges, caused mainly by Plas-modium falciparum and Plasmodium vivax. During the invasion process, the P. falciparum reticulo-cyte-binding homologous proteins PfRH1, PfRH2a, PfRH2b, PfRH4 and PfRH5 are involved, which after binding to specific membrane receptors allow the invasion of the merozoite into the erythrocyte. Objective: To compile and summarize the molecular and structural characteristics of the interactions between proteins belonging to the P. falciparum family of reticulocyte-binding homologous proteins and the receptors expressed in the host cell. Method: Descriptive review of the P. falciparum reticulocyte-binding homologous proteins involved in the process of erythrocyte invasion. This review includes literature published until 2020 in electronic databases specialized in biomedical research. We found 105 papers, of which 70 were selected and 11 were excluded for not presenting the inclusion criteria, analyzing a total of 59 references. Conclusion: The invasion of merozoite is mediated by specific interactions of the ligands of the LBS and PfRH families. The binding of the PfRH1 and PfRH2b proteins to their receptors in the erythrocyte results in the release of the EBL-175 protein, which together with PfRH4 mediates the formation of a close bond between the parasite and the red blood cells, thus allowing the binding of the PfRH5 protein to basigin and the entry of the parasite into the host cell.


Introdução: a malária é um dos maiores desafios globais de saúde pública. É causada principalmente pelos parasitas Plasmodium falciparum e Plasmodium vivax. Durante o processo de invasão, proteínas homólogas de ligação a reticulócitos de P. falciparum PfRH1, PfRH2a, PfRH2b, PfRH4 e PfRH5 estão envolvidas, que após a ligação a receptores de membrana específicos permitem a invasão do mero-zoíta ao andritro. Objetivo: compilar e resumir as características moleculares e estruturais das interações entre as pro-teínas pertencentes à família das proteínas reticulocitárias homólogas de P. falciparum e os receptores expressos na célula hospedeira. Método: revisão descritiva das proteínas ho-mólogas de ligação a reticulócitos de P. falciparum envol-vidas no processo de invasão eritrocitária. Esta revisão inclui literatura publicada até 2020 em bases de dados eletrônicas especializadas em pesquisa biomédica. Foram encontrados 105 documentos, dos quais 70 foram selecionados e 11 excluídos por não apresentarem os critérios de inclusão, anali-sando um total de 59 referências. Conclusão: a invasão de merozoítos é mediada por interações específicas dos ligantes das famílias EBL e PfRH. A ligação das proteínas PfRH1 e PfRH2b aos seus receptores no eritrócito resulta na libe-ração da prote-ína EBL-175 que, junto com PfRH4, a mediação da formação de uma junção compacta entre o parasita e as hemácias. Isso permite a ligação da proteína PfRH5 à basigina e a entrada do parasita na célula hospedeira.


Subject(s)
Malaria , Plasmodium falciparum , Erythrocyte Membrane , Erythrocytes , Ligands
3.
Journal of Experimental Hematology ; (6): 1826-1830, 2020.
Article in Chinese | WPRIM | ID: wpr-879978

ABSTRACT

OBJECTIVE@#To investigate the regulatory effects of Olaparib on natural killer cell activating receptor (NKG2D) ligands expression on human acute myeloid leukemia (AML) cell line HL-60, and to explore the molecular mechanism of Olaparib on HL-60 cells.@*METHODS@#After HL-60 cells in logarithmic growth phase were treated with Olaparib at different concentrations for different times (24, 48 h), the expression of NKG2D ligand on the surface of HL-60 cells was detected by flow cytometry. Western blot was used to dectect the expression of ERK expression in HL-60 cells. The killing effect of NK cells to HL-60 cells was detected by CFSE/PI method.@*RESULTS@#10 μmol/L Olaparib could upregulate the expression of NKG2D ligand on the surface of HL-60 cell at 24 and 48 hours, while 5 μmol/L Olaparib could induce up-regulation of the expression of ULBP-2 and ULBP-3 at 48 hours. Western blot analysis showed that ERK phosphorylation of HL-60 cells was enhanced after treating with Olaparib. The killing effect of NK cells to HL-60 cells could be enhanced by Olaparib, however, ERK inhibitor could suppress the killing effect of NK cells to HL-60 cells.@*CONCLUSION@#Olaparib can upregulate NKG2D ligands expression on the surface of HL-60 cells and enhance the cytotoxicity of NK cell to HL-60 cells. The mechanism may be related to Olaparib promoting ERK phosphorylation expression.


Subject(s)
Cell Line, Tumor , Cytotoxicity, Immunologic , HL-60 Cells , Histocompatibility Antigens Class I , Humans , Ligands , NK Cell Lectin-Like Receptor Subfamily K , Phthalazines , Piperazines , Poly(ADP-ribose) Polymerase Inhibitors
4.
Immune Network ; : 5-2020.
Article in English | WPRIM | ID: wpr-811177

ABSTRACT

The γδ T cells are unconventional lymphocytes that function in both innate and adaptive immune responses against various intracellular and infectious stresses. The γδ T cells can be exploited as cancer-killing effector cells since γδ TCRs recognize MHC-like molecules and growth factor receptors that are upregulated in cancer cells, and γδ T cells can differentiate into cytotoxic effector cells. However, γδ T cells may also promote tumor progression by secreting IL-17 or other cytokines. Therefore, it is essential to understand how the differentiation and homeostasis of γδ T cells are regulated and whether distinct γδ T cell subsets have different functions. Human γδ T cells are classified into Vδ2 and non-Vδ2 γδ T cells. The majority of Vδ2 γδ T cells are Vγ9δ2 T cells that recognize pyrophosphorylated isoprenoids generated by the dysregulated mevalonate pathway. In contrast, Vδ1 T cells expand from initially diverse TCR repertoire in patients with infectious diseases and cancers. The ligands of Vδ1 T cells are diverse and include the growth factor receptors such as endothelial protein C receptor. Both Vδ1 and Vδ2 γδ T cells are implicated to have immunotherapeutic potentials for cancers, but the detailed elucidation of the distinct characteristics of 2 populations will be required to enhance the immunotherapeutic potential of γδ T cells. Here, we summarize recent progress regarding cancer immunology of human γδ T cells, including their development, heterogeneity, and plasticity, the putative mechanisms underlying ligand recognition and activation, and their dual effects on tumor progression in the tumor microenvironment.


Subject(s)
Allergy and Immunology , Communicable Diseases , Cytokines , Homeostasis , Humans , Interleukin-17 , Ligands , Lymphocytes , Mevalonic Acid , Plastics , Population Characteristics , Protein C , Receptors, Antigen, T-Cell, gamma-delta , Receptors, Growth Factor , T-Lymphocyte Subsets , T-Lymphocytes , Terpenes , Tumor Microenvironment
5.
Immune Network ; : 11-2020.
Article in English | WPRIM | ID: wpr-811171

ABSTRACT

Most patients with hepatocellular carcinoma (HCC) are diagnosed at an advanced stage of disease. Until recently, systemic treatment options that showed survival benefits in HCC have been limited to tyrosine kinase inhibitors, antibodies targeting oncogenic signaling pathways or VEGF receptors. The HCC tumor microenvironment is characterized by a dysfunction of the immune system through multiple mechanisms, including accumulation of various immunosuppressive factors, recruitment of regulatory T cells and myeloid-derived suppressor cells, and induction of T cell exhaustion accompanied with the interaction between immune checkpoint ligands and receptors. Immune checkpoint inhibitors (ICIs) have been interfered this interaction and have altered therapeutic landscape of multiple cancer types including HCC. In this review, we discuss the use of anti-PD-1, anti-PD-L1, and anti-CTLA-4 antibodies in the treatment of advanced HCC. However, ICIs as a single agent do not benefit a significant portion of patients. Therefore, various clinical trials are exploring possible synergistic effects of combinations of different ICIs (anti-PD-1/PD-L1 and anti-CTLA-4 antibodies) or ICIs and target agents. Combinations of ICIs with locoregional therapies may also improve therapeutic responses.


Subject(s)
Antibodies , Carcinoma, Hepatocellular , Humans , Immune System , Immunotherapy , Ligands , Protein-Tyrosine Kinases , Receptors, Vascular Endothelial Growth Factor , T-Lymphocytes, Regulatory , Tumor Microenvironment
6.
Article in Chinese | WPRIM | ID: wpr-828487

ABSTRACT

OBJECTIVE@#To provide data support for the study of pathogenic mechanism of SARS-CoV-2 at the molecular level, and provide suitable candidate targets for vaccine, antibody and drug research and development through comparative analysis for structural characteristics and epitopes of S protein of SARS-CoV-2 and SARS-CoV.@*METHODS@#Based on the reference sequences of S protein, physical and chemical properties, hydrophobicity, signal peptide, transmembrane region, domain, secondary structure, tertiary structure analysis and antigenic epitopes prediction were carried out. Meanwhile, the tissue expression, related pathways and reactome pathways of angiotensis Ⅰ converting enzyme 2 (ACE2) and C-type lectin domain family 4 member M (CLEC4M) receptors were analyzed.@*RESULTS@#The amino acid sequence of S protein of SARS-CoV-2 and SARS-CoV has a 75.80% consistency. The structural characteristics of the two coronaviruses are highly consistent, but the secondary structure and tertiary structure of SARS-CoV-2 is not as obvious as SARS-CoV. ACE2 and CLEC4M are expressed in alimentary system, heart, kidney, lung and placenta. The main related the pathways of renin-angiotensin system, protein digestion and absorption pathway, and the reactome pathways of metabolism of angiotensinogen to angiotensins, GPCR ligand binding, are related to typical symptoms of coronavirus disease 2019 induced by SARS-CoV-2. Three pairs of highly or completely homologous epitopes of S protein were obtained. The 600-605, 695-703 and 888-896 amino acid residues in SARS-CoV-2 were highly homologous with 586-591, 677-685 and 870-878 amino acid residues in SARS-CoV, respectively.@*CONCLUSIONS@#The similarity of S protein of SARS-CoV-2 and SARS-CoV determines that they have similar infection patterns and clinical manifestations. The candidate epitopes with high reliability can provide reference for virus diagnosis and vaccine development.


Subject(s)
Betacoronavirus , Cell Adhesion Molecules , Coronavirus Infections , Epitopes , Humans , Lectins, C-Type , Ligands , Pandemics , Peptidyl-Dipeptidase A , Pneumonia, Viral , Receptors, Cell Surface , Receptors, Virus , Reproducibility of Results , Spike Glycoprotein, Coronavirus
7.
Article in Chinese | WPRIM | ID: wpr-828368

ABSTRACT

G-quadruplex DNA has become an important target for tumor therapy and anti-tumor development. Modern pharmacology has proved that Macleaya cordata has anti-inflammatory, antibacterial, anti-tumor and other pharmacological effects. Affinity ultrafiltration method can screen active ingredients from compounds rapidly, but G-quadruplex DNA ligands are difficult to dissociate, which is a key step in conventional ultrafiltration method. In this paper, the filtrates after ultrafiltration were determined by HPLC-MS in substitution. The peaks with 20% reduction of MS response from the incubation vs control were considered to be ligand components to G-quadruplex. Two of the peaks with the relative abundance above 30% were identified as sanguinarine(SAN) and chelerine(CHE). Their circular dichroism conformations further proved that SAN and CHE are active ligands of HT4. In addition, another two gradients with high relative abundance were identified as protopine(PRO) and allpcryprotopine(ALL). The binding rate of SAN, CHE, PRO and ALL was calculated according to the HPLC-MS results, and the results showed a consistency with that of the molecular docking method. The proposed method can be used to screen active components from mixture.


Subject(s)
Chromatography, High Pressure Liquid , Chromatography, Liquid , G-Quadruplexes , Ligands , Mass Spectrometry , Molecular Docking Simulation , Ultrafiltration
8.
Article in English | WPRIM | ID: wpr-787281

ABSTRACT

Scavenger receptors typically bind to multiple ligands on a cell surface, including endogenous and modified host-derived molecules and microbial pathogens. They promote the elimination of degraded or harmful substances such as non-self or altered-self targets through endocytosis, phagocytosis, and adhesion. Currently, scavenger receptors are subdivided into eight classes based on several variations in their sequences due to alternative splicing. Since recent studies indicate targeting scavenger receptors has been involved in cancer prognosis and carcinogenesis, we will focus on the current knowledge about the emerging role of scavenger receptor classes A to E in cancer progression.


Subject(s)
Alternative Splicing , Carcinogenesis , Endocytosis , Ligands , Macrophages , Phagocytosis , Prognosis , Receptors, Scavenger
9.
Article in English | WPRIM | ID: wpr-787280

ABSTRACT

Cancer remains a leading cause of death, despite multimodal treatment approaches. Even in patients with a healthy immune response, cancer cells can escape the immune system during tumorigenesis. Cancer cells incapacitate the normal cell-mediated immune system by expressing immune modulation ligands such as programmed death (PD) ligand 1, the B7 molecule, or secreting activators of immune modulators. Chimeric antigen receptor (CAR) T cells were originally designed to target cancer cells. Engineered approaches allow CAR T cells, which possess a simplified yet specific receptor, to be easily activated in limited situations. CAR T cell treatment is a derivative of the antigen-antibody reaction and can be applied to various diseases. In this review, the current successes of CAR T cells in cancer treatment and the therapeutic potential of CAR T cells are discussed.


Subject(s)
Antigen-Antibody Reactions , Carcinogenesis , Cause of Death , Cell- and Tissue-Based Therapy , Combined Modality Therapy , Humans , Immune System , Ligands , Receptors, Antigen , T-Lymphocytes , United Nations
10.
Con-ciencia (La Paz) ; 7(2): 55-72, nov. 2019. ilus.
Article in Spanish | LILACS | ID: biblio-1178630

ABSTRACT

El acoplamiento molecular es un método bioinformático que permite predecir y calcular computacionalmente la posición más favorable de interacción entre un ligando y un blanco (usualmente proteico) a partir de sus representaciones tridimensionales. Esta herramienta bioinformática no tiene una regla que se adapte a todos los casos y la mayoría de los programas empleados con esta finalidad, tienen diferentes métodos para tratar cada caso en particular. Además, cada blanco proteico es estructuralmente diferente y la capacidad de replicar los resultados experimentales y fisiológicos depende en gran medida del sistema utilizado y del criterio del usuario. Así, este artículo proporcionará pautas y detallará varios aspectos prácticos como el estado de protonación de los átomos, la identificación de moléculas de agua importantes en el sitio activo, la diferenciación entre moléculas activas y señuelos como mecanismo de control, la determinación de la energía libre de unión (∆G) y el análisis de la flexibilidad y la dinámica molecular; aspectos que deberían ser tomados en cuenta antes de iniciar un estudio de acoplamiento molecular para la búsqueda y selección virtual de ligandos e identificación de blancos terapéuticos.


Molecular docking is a bioinformatics method that allows predicting and calculating computationally the most favorable position of interaction between a ligand and a target (usually a protein) from its three-dimensional representations. This bioinformatics tool does not have a rule that suits all cases and most of the programs used for this purpose have different methodologies to deal with each particular case. In addition, each protein target is structurally different and the ability to replicate the experimental and physiological results depends largely on the system used and the user's criteria. Thus, this article will provide guidelines and detail several practical aspects such as the protonation state of atoms, the identification of important water molecules in the active site, the differentiation between active molecules and decoys as a control mechanism, the determination of free energy binding (∆G) and analysis of flexibility and molecular dynamics; aspects that should be taken into account before starting a molecular docking study for a virtual ligand screening and identification of therapeutic targets.


Subject(s)
Weights and Measures , Computational Biology , Ligands , Control
11.
Article in Chinese | WPRIM | ID: wpr-773224

ABSTRACT

Inflammatory response is caused by exogenous and endogenous stimuli,resulting in a non-specific resistance reaction.After acute ischemic cerebral infarction,inflammatory factors gather and adhere in the ischemic area of leukocyte infiltration,and the released inflammatory factors causes the injury cascade,aggravate the brain tissue damage and the symptoms of neurological deficits,and hinder the repair of brain neurons and the recovery of nerve function. In this paper,the key targets in the arachidonic acid metabolic pathway were studied. The Hiphop pharmacophore model of s PLA2-ⅡA and COX-2 inhibitors was built. According tothe two previously constructed 5-LOX and LTA4 H target inhibitors,the pharmacophore model was used to initially screen out the composition database of all of 13 traditional Chinese medicines in Xixian Tongshuan Preparation. The molecular matching study was carried out by selecting the matching value greater than 0. 6,and the component with the CDOCKER score greater than 80% of the original ligand score was used as the potential active inhibitor of the target. Considering the pharmacophore matching value,the molecular docking score and the interaction between the components and the target,one Chuanxiong component and one safflower component were selected as potential inhibitors of s PLA2-ⅡA; two Chuanxiong components,two Panax notoginseng,one safflower component,one angelica component,one valerian component were taken as a potential inhibitor of COX-2; two Gentiana components,one safflower component,one valerian component,one P. notoginseng component and one Angelica component were taken as potential inhibitors of 5-LOX; and two Gentiana components,two Chuanxiong components,and two safflower components were taken as potential inhibitors of LTA4 H. This study screened out the potential inhibitors of the four targets in a high-efficiency and low-cost manner,and explained that Xixian Tongshuan Preparation showed an effect in the treatment of inflammatory responses caused by ischemic stroke by acting both LOX pathway and COX pathway in the metabolic pathway.


Subject(s)
Anti-Inflammatory Agents , Pharmacology , Drugs, Chinese Herbal , Pharmacology , Ligands , Medicine, Chinese Traditional , Molecular Docking Simulation
12.
Chinese Journal of Biotechnology ; (12): 216-225, 2019.
Article in Chinese | WPRIM | ID: wpr-771385

ABSTRACT

The mucosae represent the first line of defense against the invasion of most pathogens, and the mucosal immune system plays a crucial role in the control of infection. Mucosal vaccination can trigger both humoral and cell-mediated immune responses mucosally as well as systemically. Hence, protective immune responses can be elicited effectively by mucosal vaccination. Microfold (M) cells being unique to the mucosal immune system can take up luminal antigens and initiating antigen-specific immune responses. The number of antigen uptake by M cells is directly related to the immune efficacy of mucosal vaccines. Utilizing M cell ligands, M cells-targeting antigen delivery can achieve highly effective mucosal immune responses. The strategy of targeted delivery of antigens to M cells and its applications can be used for the improvement of mucosal immune responses and the development of mucosal vaccines. Despite these efforts, successful development of safe and effective mucosal vaccines remains a big challenge and needs a long way to go, and provably still resort to further researches on cellular properties and functions as well as mucosal immune mechanisms.


Subject(s)
Immunity, Mucosal , Ligands , Mucous Membrane , Vaccination , Vaccines , Allergy and Immunology
13.
Chinese Journal of Biotechnology ; (12): 513-521, 2019.
Article in Chinese | WPRIM | ID: wpr-771356

ABSTRACT

To explore the immobilization of target proteins for screening libraries of ligand mixtures, magnetic submicron particles (MSP) functionalized with Ni²⁺-NTA and carboxyl were compared for the immobilization of Mycobacterium tuberculosis dihydrofolate reductase (MtDHFR). MtDHFR fused with 6×His was expressed, purified and characterized for kinetics. MtDHFR was immobilized on Ni²⁺-NTA-functionalized MSP directly and carboxyl-functionalized MSP upon activation. The immobilization capacity, residual activity, thermostability and affinities for putative inhibitors were characterized. MtDHFR immobilized on Ni²⁺-NTA-functionalized MSP retained about 32% activity of the free one with the immobilization capacity of (93±12) mg/g of MSP (n=3). Ni²⁺ and EDTA synergistically inhibited MtDHFR activity, while Fe³⁺ had no obvious interference. MtDHFR immobilized on carboxyl-functionalized MSP retained (87±4)% activity of the free one with the immobilization capacity of (8.6±0.6) mg/g MSP (n=3). In 100 mmol/L HEPES (pH 7.0) containing 50 mmol/L KCl, there was no significant loss of the activities of the free and immobilized MtDHFR after storage at 0 °C for 16 h, but nearly 60% and 35% loss of their activities after storage at 25 °C for 16 h, respectively. The inhibition effects of methotrexate on the immobilized and free MtDHFR were consistent (P>0.05). The immobilization of MtDHFR on carboxyl-functionalized MSP was thus favorable for higher retained activity and better thermostability, with promise for rapid screening of its ligand mixtures.


Subject(s)
Enzyme Stability , Enzymes, Immobilized , Hydrogen-Ion Concentration , Kinetics , Ligands , Magnetite Nanoparticles , Mycobacterium tuberculosis , Temperature , Tetrahydrofolate Dehydrogenase
14.
Article in Chinese | WPRIM | ID: wpr-776034

ABSTRACT

Objective To investigate the effects of different inflammatory factors on hepatocyte kinase receptor(Eph)and ligand(ephrin)in human periodontal ligament fibroblasts(hPDLFs).Methods hPDLFs were stimulated with either 10 ng/ml tumor necrosis factor-α(TNF-α)or 10 ng/ml interleukin(IL)-1β,and then the expressions of Eph and ephrin at both mRNA and protein levels were determined at 0,1,2,6,12,and 24 hours.Results The levels of Eph receptors and ephrin ligand changed in a time-dependent manner in human periodontal ligament fibroblasts after treatment with TNF-α or IL-1β. The expression of ephrinA2 significantly increased in both groups within 24 hours(all <0.05). In the TNF-α group,the mRNA expression of ephrinA2 significantly increased at 1 h and was significant higher that in the IL-1β group at 24 h(<0.05). EphB4 showed a time-dependent decline after a short period of high expression.Conclusions Both TNF-α and IL-1β can cause changes in the expressions of Eph receptors and ephrin ligands in hPDLFs. The changes induced by both are consistent,although the effect of TNF-α is more pronounced.


Subject(s)
Cells, Cultured , Ephrins , Metabolism , Fibroblasts , Humans , Interleukin-1beta , Pharmacology , Ligands , Periodontal Ligament , Cell Biology , Receptors, Eph Family , Metabolism , Tumor Necrosis Factor-alpha , Pharmacology
15.
Asian Spine Journal ; : 875-889, 2019.
Article in English | WPRIM | ID: wpr-785500

ABSTRACT

STUDY DESIGN: Development of an in vitro model for assessing the anti-inflammatory efficacies of naringin (Nar) and naringenin (NG).PURPOSE: To evaluate the efficacy of natural flavonoids as therapeutic drugs against anti-inflammatory processes in the nucleus pulposus (NP) cells using in-vitro and in-silico methods.OVERVIEW OF LITERATURE: Intervertebral disc (IVD) disease is a common cause of low back pain. Chronic inflammation and degeneration play a significant role in its etiopathology. Thus, a better understanding of anti-inflammatory agents and their role in IVD degeneration and pro-inflammatory cytokines expression is necessary for pain management and regeneration in IVD.METHODS: We performed primary cell culture of NP cells; immunocytochemistry; gene expression studies of cytokines, metalloproteases, extracellular proteins, and apoptotic markers using quantitative polymerase chain reaction and reverse transcription-polymerase chain reaction (RT-PCR); cytotoxicity assay (MTT); and molecular docking studies using AutoDock 4.2 software (Molecular Graphics Laboratory, La Jolla, CA, USA) to confirm the binding mode of proteins and synthesized complexes. We calculated the mean±standard deviation values and performed analysis of variance and t-test using SPSS ver. 17.0 (SPSS, Inc., Chicago, IL, USA).RESULTS: Molecular docking showed that both Nar and NG bind to the selected genes of interest. Semi-quantitative RT-PCR analysis reveals differential gene expression of collagen (COL)9A1, COL9A2, COL9A3, COL11A2, COMT (catechol-O-methyltransferase), and THBS2 (thrombospondin 2); up regulation of ACAN (aggrecan), COL1A1, COL11A1, interleukin (IL)6, IL10, IL18R1, IL18RAP, metalloprotease (MMP)2, MMP3, MMP9, ADAMTS5 (a disintegrin and metalloproteinase with thrombospondin motifs 5), IGF1R (insulin-like growth factor type 1 receptor), SPARC (secreted protein acidic and cysteine rich), PARK2 (parkin), VDR (vitamin D receptor), and BCL2 (B-cell lymphoma 2); down regulation of IL1A, CASP3 (caspase 3), and nine genes with predetermined concentrations of Nar and NG.CONCLUSIONS: The present study evaluated the anti-inflammatory and regenerative efficiencies of Nar and NG in degenerated human NP cells. Altered gene expressions of cytokines, metalloproteases, extracellular proteins, apoptotic genes were dose responsive. The molecular docking (in silico) studies showed effective binding of these native ligands (Nar and NG) with genes identified as potent inhibitors of inflammation. Thus, these natural flavonoids could serve as anti-inflammatory agents in the treatment of low back pain and sciatica.


Subject(s)
Anti-Inflammatory Agents , Caspase 3 , Collagen , Cysteine , Cytokines , Down-Regulation , Flavonoids , Gene Expression , Humans , Immunohistochemistry , In Vitro Techniques , Inflammation , Interleukin-10 , Interleukins , Intervertebral Disc , Intervertebral Disc Degeneration , Ligands , Low Back Pain , Lymphoma , Metalloproteases , Models, Molecular , Pain Management , Polymerase Chain Reaction , Primary Cell Culture , Regeneration , Sciatica , Thrombospondins , Up-Regulation
16.
Article in English | WPRIM | ID: wpr-739669

ABSTRACT

BACKGROUND: Glioblastoma multiforme (GBM) is a highly malignant brain tumor with a worst prognosis of less than one year despite advance treatment facilities. Among various signaling pathway genes displaying genetic modifications, aberrant expression of Notch pathway genes is frequent in GBM offering novel therapeutic targets. Herbal extracts having anticancer properties are used in adjuvant therapy that is safe and affordable as compared to chemotherapeutics. Bacopa monnieri has been used for the development of brain cells because of its neuroprotective properties. Its anticancer properties have shown to be promising in cancer treatment. METHODS: The anticancer properties of Bacoside A, an active and abundant component of Bacopa monnieri was assessed on U-87 MG cell line and its effects on expression of Notch pathway genes were studied. Cell cycle arrest and apoptosis were studied using flow cytometry. Expression of Notch pathway genes comprising of Notch receptors (notch1, notch2, notch3 and notch4), ligands (jagged1 and jagged2), a component of gamma-secretase complex (APH1A) and downstream target (HES1) were evaluated by quantitative real-time PCR. RESULTS: Bacoside A exhibited considerable cytotoxicity on U-87 MG cells inducing cell cycle arrest and apoptosis. Cell cycle analysis revealed a significant arrest of 39.21% cells in sub-G0 phase at 80 µg/mL concentration, increasing to 53.21% at a higher concentration of 100 µg/mL. The fraction of early apoptotic cells in control was low (3.48%) that increased substantially to 31.36% and 41.11% after 80 µg/mL and 100 µg/mL of Bacoside A treatment respectively. Additionally, the expression of notch1 gene decreased after exposure to Bacoside A with a fold change of 0.05, whereas HES1 gene expression was increased by 25 fold. CONCLUSION: These data indicate that Bacoside A has a possible anticancer activity that could be inducing cell cycle arrest and apoptosis through Notch pathway in GBM in vitro.


Subject(s)
Amyloid Precursor Protein Secretases , Apoptosis , Bacopa , Brain , Brain Neoplasms , Cell Cycle , Cell Cycle Checkpoints , Cell Line , Flow Cytometry , Gene Expression , Glioblastoma , Humans , In Vitro Techniques , Ligands , Prognosis , Real-Time Polymerase Chain Reaction , Receptors, Notch
17.
Article in English | WPRIM | ID: wpr-739657

ABSTRACT

Mixed lineage leukemia proteins (MLL) are the key histone lysine methyltransferases that regulate expression of diverse genes. Aberrant activation of MLL promotes leukemia as well as solid tumors in humans, highlighting the urgent need for the development of an MLL inhibitor. We screened and isolated MLL1-binding ssRNAs using SELEX (Systemic Evolution of Ligands by Exponential enrichment) technology. When sequences in sub-libraries were obtained using next-generation sequencing (NGS), the most enriched aptamers—APT1 and APT2—represented about 30% and 26% of sub-library populations, respectively. Motif analysis of the top 50 sequences provided a highly conserved sequence: 5′-A[A/C][C/G][G/U][U/A]ACAGAGGG[U/A]GG[A/C] GAGUGGGU-3′. APT1, APT2, and APT5 embracing this motif generated secondary structures with similar topological characteristics. We found that APT1 and APT2 have a good binding activity and the analysis using mutated aptamer variants showed that the site information in the central region was critical for binding. In vitro enzyme activity assay showed that APT1 and APT2 had MLL1 inhibitory activity. Three-dimensional structure prediction of APT1-MLL1 complex indicates multiple weak interactions formed between MLL1 SET domain and APT1. Our study confirmed that NGS-assisted SELEX is an efficient tool for aptamer screening and that aptamers could be useful in diagnosis and treatment of MLL1-mediated diseases.


Subject(s)
Aptamers, Nucleotide , Conserved Sequence , Diagnosis , Histones , Humans , In Vitro Techniques , Leukemia , Ligands , Lysine , Mass Screening , Methyltransferases , Myeloid-Lymphoid Leukemia Protein , RNA
18.
Article in English | WPRIM | ID: wpr-764300

ABSTRACT

BACKGROUND: The roots of Scutellaria baicalensis Georgi (Labiatae) have been widely used in traditional medicine for treatment of various diseases. In this study, we investigated the effects of ethanol extracts of S. baicalensis roots (EESB) on the growth ofn human leukemia U937 cells. METHODS: The effect of EESB on cell viability was measured by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay. Apoptosis was determined using 4,6-diamidino-2-phenyllindile staining and flow cytometry. The effects of EESB on the expression of regulatory proteins of apoptosis and phosphatidyl inositol 3-kinase (PI3K)/Akt signaling were determined by Western blotting. Caspase activity and mitochondrial membrane potential (MMP) were measured using flow cytometric analysis.


Subject(s)
Apoptosis , Blotting, Western , Caspase 8 , Caspase 9 , Cell Survival , Down-Regulation , Ethanol , Flow Cytometry , Humans , Leukemia , Ligands , Medicine, Traditional , Membrane Potential, Mitochondrial , Phosphatidylinositols , Receptors, Death Domain , Scutellaria baicalensis , Scutellaria , U937 Cells , Up-Regulation
19.
Article in English | WPRIM | ID: wpr-763039

ABSTRACT

The capsaicin receptor TRPV1 (transient receptor potential vanilloid 1) has been an object of intense interest for pharmacological development on account of its critical role in nociception. In the course of structure activity analysis, it has become apparent that TRPV1 ligands may vary dramatically in the rates at which they interact with TRPV1, presumably reflecting differences in their abilities to penetrate into the cell. Using a fast penetrating agonist together with an excess of a slower penetrating antagonist, we find that we can induce an agonist response of limited duration and, moreover, the duration of the agonist response remains largely independent of the absolute dose of agonist, as long as the ratio of antagonist to agonist is held constant. This general approach for limiting agonist duration under conditions in which absolute agonist dose is variable should have more general applicability.


Subject(s)
Capsaicin , Ligands , Nociception
20.
Article in English | WPRIM | ID: wpr-761927

ABSTRACT

BACKGROUND: For the bone-specific imaging, a structure-inherent targeting of bone tissue recently has been reported a new strategy based on incorporation of targeting moieties into the chemical structure of near-infrared (NIR) contrast agents, while conventional methods require covalent conjugation of bone-targeting ligands to NIR contrast agents. This will be a new approach for bone-targeted imaging by using the bifunctional NIR contrast agents. METHODS: The goal of this review is to provide an overview of the recent advances in optical imaging of bone tissue, highlighting the structure-inherent targeting by developing NIR contrast agents without the need for a bone-targeting ligand such as bisphosphonates. RESULTS: A series of iminodiacetated and phosphonated NIR contrast agents for the structure-inherent targeting of bone tissue showed excellent bone-targeting ability in vivo without non-specific binding. Additionally, the phosphonated NIR contrast agents could be useful in the diagnosis of bone metastasis. CONCLUSION: By developing bone-targeted NIR contrast agents, optical imaging of bone tissue makes it very attractive for preclinical studies of bone growth or real-time fluorescence guided surgery resulting in high potential to shift the clinical paradigms.


Subject(s)
Bone and Bones , Bone Development , Contrast Media , Diagnosis , Diphosphonates , Fluorescence , Ligands , Neoplasm Metastasis , Optical Imaging , Surgery, Computer-Assisted
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