Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 6.432
Filtrar
1.
Gene ; 732: 144336, 2020 Mar 30.
Artículo en Inglés | MEDLINE | ID: mdl-31935514

RESUMEN

In the present study, we aimed to evaluate effects of autologous mesenchymal stem cells (MSCs) intravenous administration on the response of B cells, BAFF, APRIL, and their receptors on the surface of B cells at 1, 6, and 12 month follow-up periods in refractory rheumatoid arthritis (RA) patients. Thirteen patients with refractory RA received autologous MSCs. Plasma levels of BAFF and APRIL were measured employing ELISA method, followed by estimating B cell population and BAFFRs evaluation by flow cytometry technique. Gene expression of BAFF, APRIL, and their receptors on B cell surface in PBMCs was evaluated by SYBR Green real-time PCR technique. Plasma concentration of BAFF significantly decreased 1 and 6 months after the MSCT (MSCs Transplantation). Plasma concentration of APRIL significantly decreased 1 month after the MSCT. Percentages of CD19 + B cells in the PBMC population significantly decreased 12 months after the MSCT. Percentages of BR3 + CD19 + B cells and BCMA + CD19 + B cells significantly decreased at the 12th month after the MSCT. The gene expression of BAFF in the PBMC population significantly decreased during 6, and 12 months after the MSCT. The gene expression of APRIL significantly decreased on month 6 after the MSCT. The gene expression of BR3 significantly decreased during 1, 6, and 12 months after the MSCT. The MSCT seems to decrease B cells response because of the reduced production of BAFF and APRIL cytokines and decrease the expression of their receptors on the surface of B cells.


Asunto(s)
Artritis Reumatoide/terapia , Factor Activador de Células B/metabolismo , Receptor del Factor Activador de Células B/metabolismo , Regulación hacia Abajo , Trasplante de Células Madre Mesenquimatosas/métodos , Miembro 13 de la Superfamilia de Ligandos de Factores de Necrosis Tumoral/metabolismo , Administración Intravenosa , Adulto , Antígenos CD19/metabolismo , Artritis Reumatoide/genética , Artritis Reumatoide/inmunología , Factor Activador de Células B/genética , Receptor del Factor Activador de Células B/genética , Linfocitos B/inmunología , Femenino , Perfilación de la Expresión Génica , Humanos , Persona de Mediana Edad , Resultado del Tratamiento , Miembro 13 de la Superfamilia de Ligandos de Factores de Necrosis Tumoral/genética
2.
Cell Prolif ; 53(1): e12722, 2020 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-31737959

RESUMEN

OBJECTIVES: The mechanisms underlying the effects of Toll-like receptor 9 (TLR9) and autophagy on rheumatoid arthritis (RA)-aggravated periodontitis are unclear. We aimed to explore a novel target, cathepsin K (Ctsk)-mediated TLR9-related autophagy, during the progress of periodontitis with RA. MATERIALS AND METHODS: DBA/J1 mouse model of periodontitis with RA was created by local colonization of Porphyromonas gingivalis (Pg) and injection of collagen. The expression of Ctsk was inhibited by adeno-associated virus (AAV). Micro-CT, immunohistochemistry (IHC), Western blot and quantitative real-time polymerase chain reaction (qRT-PCR) were used to detect the expression of TLR9-related autophagy in periodontitis with RA. Small interfering RNA (siRNA) and CpG oligodeoxynucleotides (CpG ODN) were applied in macrophages. Western blot, immunofluorescence (IF) and qRT-PCR were used to verify the in vivo results. RESULTS: RA can promote periodontitis bone destruction in the lesion area, while inhibiting Ctsk could effectively alleviate this effect. The infiltration of macrophages, TLR9, autophagy proteins (TFEB and LC3) and inflammatory cytokines increased in the periodontitis-with-RA group and was reduced by the inhibition of Ctsk in the periodontal region. Macrophage stimulation confirmed the in vivo results. With the activation of TLR9 by CpG ODN, inhibition of Ctsk could suppress both TLR9 downstream signalling proteins and autophagy-related proteins. CONCLUSIONS: This study advanced a novel role for Ctsk in TLR9 and autophagy to explain the interaction between periodontitis and RA.


Asunto(s)
Artritis Reumatoide/tratamiento farmacológico , Catepsina K/antagonistas & inhibidores , Regulación hacia Abajo/efectos de los fármacos , Oligodesoxirribonucleótidos/farmacología , Periodontitis/tratamiento farmacológico , Receptor Toll-Like 9/inmunología , Animales , Artritis Reumatoide/genética , Artritis Reumatoide/inmunología , Artritis Reumatoide/patología , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/genética , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/inmunología , Catepsina K/genética , Catepsina K/inmunología , Modelos Animales de Enfermedad , Masculino , Ratones , Ratones Endogámicos DBA , Proteínas Asociadas a Microtúbulos/genética , Proteínas Asociadas a Microtúbulos/inmunología , Oligodesoxirribonucleótidos/genética , Periodontitis/genética , Periodontitis/inmunología , Periodontitis/patología , Receptor Toll-Like 9/genética
4.
Gene ; 722: 144098, 2020 Jan 05.
Artículo en Inglés | MEDLINE | ID: mdl-31494241

RESUMEN

This study evaluated the possible association between SNPs in cytokines coding genes, namely IL10, IL6 and IFNG, cytokines serum levels and clinical assessment' scores in patients with Rheumatoid Arthritis(RA). SNPs genotyping was performed in 126 RA patients and 177 healthy individuals with Taqman probes specific for IL10 -1082 (T>C, rs1800896);INFG -1616 (A>G, rs2069705) and IL6 -174 (G>C, rs1800795) variants,positioned in regulatory regions. Cytokine Bead Array (CBA) was used to measure cytokine levels. We found association between INFG -1616 G allele(p = 0.0210; OR = 1.605) and INFG -1616 GG genotype (p = 0.0268; OR =2.609) and RA susceptibility. We also observed association between IL10 -1082 TT genotype and high clinical disease activity index (CDAI) values (p = 0.026; OR = 1.906; 95% CI = 1.082 - 3.359), IL10 -1082 CC genotype and low CDAI values (p = 0.016; OR = 0.256) and INFG -1616 AA and high CDAI values (p = 0.025; OR = 2.919). IL10 -1082 CC also exhibited the lowest IL-10 levels than IL10 -1082 TT (p = 0.020) and IL10 -1082 TC (p = 0.032). Finally, we verified higher IL-6 value in the RA patients than healthy control group (p = 0.007) and an association between high IL-6 levels and increased CDAI (r = 0.4648, p = 0.0015); DAS 28 (r = 0.3933, p= 0.0091), presence of bone erosions (r = 0.3170, p = 0.0361), ESR levels(r = 0.3041, p = 0.0448) and IFN-γ levels (r = 0.3049, p = 0.0468).Altogether, we suggest that IL10 -1082 (T>C, rs1800896) and INFG -1616(A>G, rs2069705) polymorphisms as well as IL-6 levels alterations may play a role for prognostic and disease follow-up.


Asunto(s)
Artritis Reumatoide/genética , Interferón gamma/genética , Interleucina-10/genética , Interleucina-6/genética , Polimorfismo de Nucleótido Simple , Adulto , Artritis Reumatoide/sangre , Artritis Reumatoide/diagnóstico , Femenino , Frecuencia de los Genes , Genotipo , Humanos , Interferón gamma/sangre , Interleucina-10/sangre , Interleucina-6/sangre , Masculino , Persona de Mediana Edad
5.
Gene ; 722: 144105, 2020 Jan 05.
Artículo en Inglés | MEDLINE | ID: mdl-31521702

RESUMEN

BACKGROUND: Caulophyllum robustum Maxim (CRM) is a medicinal compound of the Northeast and is commonly used in China for the treatment of rheumatic pain and rheumatoid arthritis (RA). A preliminary study found that CRM has good anti-inflammatory, analgesic and immunosuppressive effects. However, the specific links and targets for its function remain unclear. Our study aimed to provide a mechanism for the action of Caulophyllum robustum Maxim extraction (CRME) against RA and to establish a method for studying disease treatment using Chinese medicine. METHODS: The 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) method was used to detect the toxicity of CRME in L929 cells, and the concentration ranges of the blank, model, and CRME drug groups were determined. Differentially expressed long non-coding RNAs (lncRNAs) and messenger RNAs (mRNAs) were identified between the three groups. Gene Ontology (GO) and pathway enrichment analyses were performed to analyze the biological functions and pathways of the differentially expressed genes. Expression of Hist1h2bj, Hist1h2ba, Zfp36, Ccl3, Cxcl2 and Egr1 in the blank, model and drug groups was detected by quantitative real-time PCR (qRT-PCR), and the role of CRME on the above factors was determined to ensure consistency with the chip data. RESULTS: A total of 329 significantly upregulated genes and 141 downregulated genes were identified between the blank and model groups. A total of 218 significantly upregulated genes and 191 downregulated genes were identified between the CRME drug group and model group. CRME has a significant role in multiple pathways involved in the occurrence and development of RA. Additionally, Hist1h2bj, Hist1h2ba, Zfp36, Ccl3, Cxcl2, and Egr1 were observed in modules of the lncRNA-mRNA weighted co-expression network, consistent with the chip data. CONCLUSIONS: CRME has regulatory effects on inflammatory factors, the histone family, chemokines and their ligands that are related to RA-related cytokines, the RA pathway, the TNF signaling pathway, the Toll receptor-like signaling pathway, the chemokine signaling pathways and other pathways are related to the course of RA.


Asunto(s)
Artritis Reumatoide/genética , Caulophyllum , Medicamentos Herbarios Chinos/farmacología , ARN Largo no Codificante/metabolismo , ARN Mensajero/metabolismo , Animales , Artritis Reumatoide/metabolismo , Línea Celular , Expresión Génica/efectos de los fármacos , Ratones , Análisis de Secuencia por Matrices de Oligonucleótidos , Mapeo de Interacción de Proteínas
6.
Gene ; 724: 144144, 2020 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-31629819

RESUMEN

BACKGROUND/AIMS: Rheumatoid arthritis synovial fibroblasts (RASF) play an essential role in the pathogenesis of rheumatoid arthritis (RA). This study aimed to investigate the biological effects of miR-22 on RASFs. METHODS: RT-qPCR was used to detect the expressions of miR-22 and SIRT1 in RA synovial tissue. The results of miR-22 on the proliferation of RASF were examined by MTT assay. The effects of miR-22 on the secretion of TNF-α, IL-1ß, and IL-6 in RASF were measured by ELISA. Target gene prediction and screening, and luciferase reporter assay were used to testify downstream target genes of miR-22. RT-qPCR and western blotting were used to detect the mRNA and protein expression of SIRT1. RESULTS: miR-22 was significantly decreased in RA synovial tissue, while SIRT1 was significantly increased in RA synovial tissue. Over-expression of miR-22 significantly inhibited the proliferation of RASFs and the secretions of inflammatory cytokines (TNF-α, IL-1ß, and IL-6) in RASFs. SIRT1 was identified as a direct target of miR-22. Over-expression of miR-22 reduced the expression level of SIRT1 in RASFs. Over-expression of SIRT1 reversed the effect of miR-22 on the proliferation of RASFs and the secretion of inflammatory cytokines. CONCLUSION: MIR-22 was significantly down-regulated in RASF cells, which affected the secretions of inflammatory cytokines and cell proliferation by regulating SIRT1.


Asunto(s)
Artritis Reumatoide/genética , Citocinas/metabolismo , MicroARNs/genética , Sirtuina 1/genética , Membrana Sinovial/patología , Regiones no Traducidas 3' , Artritis Reumatoide/patología , Proliferación Celular/genética , Citocinas/genética , Femenino , Fibroblastos/patología , Humanos , Masculino , MicroARNs/metabolismo , Persona de Mediana Edad , Sirtuina 1/metabolismo , Membrana Sinovial/metabolismo
7.
Medicine (Baltimore) ; 98(52): e18606, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31876763

RESUMEN

The aim of the present study was to examine the association between vascular endothelial growth factor receptor 2 (VEGFR2) rs11941492 C/T polymorphism and rheumatoid arthritis (RA) risk in an eastern Chinese Han population. We examined VEGFR2 rs11941492 C/T polymorphism in 615 RA patients and 839 controls in an East Chinese Han population. The power analysis was used for evaluating the reliability of the results. Genotyping was performed using a custom-by-design 48-Plex single nucleotide polymorphism scan Kit. Pooled odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using logistic regression.Our results indicated that VEGFR2 rs11941492 C/T polymorphism (TT vs CC, P = .012, OR = 0.61, 95% CI = 0.41-0.89; TT vs CT + CC, P = .017, OR = 0.63, 95% CI = 0.43-0.92) was associated with a significantly decreased risk of RA. The power analysis showed that this study had a power of 98.5% to detect the effect of rs11941492 C/T polymorphism on RA susceptibility, assuming an OR of 0.61. After stratification analysis, a decreased risk of RA was associated with VEGFR2 rs11941492 TT genotype (TT vs CC) among female patients (TT vs CC, P = .007, OR = 0.53, 95% CI = 0.33-0.84), older patients (Yr ≥55) (TT vs CC, P = .039, OR = 0.58, 95% CI = 0.35-0.97), C-reactive protein-positive patients, anti-cyclic citrullinated peptide antibody-negative patients, rheumatoid factor-positive patients (TT vs CT + CC, P = .015, OR = 0.60, 95% CI = 0.39-0.90), functional class III + IV patients, patients with a DAS28 of ≥3.20, and those with an erythrocyte sedimentation rate of <25. However, our results were obtained from only a moderate-sized sample. Studies with larger sample sizes in other ethnic populations are needed to confirm these results. The VEGFR2 rs11941492 genotype is associated with decreased susceptibility to RA.


Asunto(s)
Artritis Reumatoide/genética , Polimorfismo de Nucleótido Simple/genética , Receptor 2 de Factores de Crecimiento Endotelial Vascular/genética , Grupo de Ascendencia Continental Asiática/genética , Estudios de Casos y Controles , China , Femenino , Predisposición Genética a la Enfermedad/genética , Humanos , Modelos Logísticos , Masculino , Persona de Mediana Edad , Oportunidad Relativa
8.
Genome Biol ; 20(1): 233, 2019 11 06.
Artículo en Inglés | MEDLINE | ID: mdl-31694669

RESUMEN

The driver tissues or cell types in which susceptibility genes initiate diseases remain elusive. We develop a unified framework to detect the causal tissues of complex diseases or traits according to selective expression of disease-associated genes in genome-wide association studies (GWASs). This framework consists of three components which run iteratively to produce a converged prioritization list of driver tissues. Additionally, this framework also outputs a list of prioritized genes as a byproduct. We apply the framework to six representative complex diseases or traits with GWAS summary statistics, which leads to the estimation of the lung as an associated tissue of rheumatoid arthritis.


Asunto(s)
Enfermedad/etiología , Expresión Génica , Predisposición Genética a la Enfermedad , Genómica/métodos , Algoritmos , Artritis Reumatoide/genética , Artritis Reumatoide/metabolismo , Trastorno Bipolar/genética , Trastorno Bipolar/metabolismo , Estatura/genética , Encéfalo/metabolismo , Colesterol/genética , Enfermedad de la Arteria Coronaria/genética , Enfermedad de la Arteria Coronaria/metabolismo , Estudio de Asociación del Genoma Completo , Humanos , Esquizofrenia/genética , Esquizofrenia/metabolismo
9.
Medicine (Baltimore) ; 98(44): e17604, 2019 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-31689765

RESUMEN

This study genotyped blood samples from 214 patients with rheumatoid arthritis (RA) and 293 healthy controls for single nucleotide polymorphisms (SNPs) rs2977537, rs2929970, rs2929973, rs2977530, rs1689334 and rs62514004. We want to investigate whether the SNPs in the WNT1-inducible signaling pathway protein 1 (WISP-1) gene may increase the risk of developing RA. We showed that RA disease was more likely with the AA genotype compared with the AG genotype of SNP rs2977537 (adjusted odds ratio [AOR]: 0.54; 95% confidence interval [CI]: 0.34-0.84), and with the TT genotype (AOR: 0.24; 95% CI: 0.13-0.39) or the GG genotype (AOR: 0.05; 95% CI: 0.03-0.10) compared with the GT genotype of rs2929973, and with the AA genotype (AOR: 0.34; 95% CI: 0.22-0.54) or GG genotype (AOR: 0.52; 95% CI: 0.31 to 0.87) vs the AG genotype of rs2977530. Rheumatoid factor positivity was more likely with the AA genotype than with the AG genotype of the rs2977537 polymorphism (AOR: 0.16; 95% CI: 0.16-0.94). High CRP (>8 mg/L) was more likely with the non-AG genotype (AA + GG) than the AG genotype of rs2977537 (AOR: 1.84; 95% CI: 1.05-3.21) and with the AA genotype vs the AG genotype of rs2977530 (AOR: 2.62; 95% CI: 1.35-5.09). Compared with the AG genotype, the AA genotype of rs2929970 was more likely to require prednisolone (AOR: 0.49; 95% CI: 0.27-0.88), while the AG genotype was more likely than the AA genotype of SNP rs2977530 to require TNF-α inhibitors (AOR: 2.07; 95% CI: 1.08 to 3.98). WISP-1 may be a diagnostic marker and therapeutic target for RA therapy.


Asunto(s)
Antirreumáticos/uso terapéutico , Artritis Reumatoide/tratamiento farmacológico , Artritis Reumatoide/genética , Prednisolona/uso terapéutico , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Anciano , Alelos , Artritis Reumatoide/etnología , Grupo de Ascendencia Continental Asiática , Biomarcadores , Proteínas CCN de Señalización Intercelular , Estudios de Casos y Controles , Femenino , Predisposición Genética a la Enfermedad , Genotipo , Humanos , Masculino , Persona de Mediana Edad , Neovascularización Patológica , Oportunidad Relativa , Polimorfismo de Nucleótido Simple , Proteínas Proto-Oncogénicas , Reacción en Cadena en Tiempo Real de la Polimerasa
10.
Medicine (Baltimore) ; 98(48): e18218, 2019 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-31770283

RESUMEN

Rheumatoid arthritis (RA) is a systemic autoimmune disease characterized by joint destructions and human leukocyte antigen (HLA)-DRB1 is an important genetic risk factor for RA and influences the phenotype of RA. The clinical features of elder age onset RA (EORA) were known to be different from those of younger age onset RA (YORA). Previous studies reported the different association pattern of DRB1 alleles with YORA or EORA. The associations of DRB1 genotype with these RA subsets remained almost unknown. We investigated the genotype association of DRB1 with YORA or EORA in Japanese populations.HLA genotyping was performed in Japanese RA patients and the association of allele or genotype carrier frequencies were analyzed.The genotype frequency of DRB104:05/DRB104:06 (P = .0204, OR 7.69, 95%CI 1.39-42.72), DRB104:05/DRB112:01 (P = .0050, OR 5.53, 95%CI 1.71-17.88), and DRB104:05/DRB115:01 (P = .0124, OR 3.34, 95%CI 1.39-8.02) in YORA was higher than EORA. However, the frequencies of DRB101:01/DRB104:05 in YORA was tended to be lower than EORA (P = .0784, OR 0.14, 95%CI 0.01-2.42). The gene dosage effect of the shared epitope alleles was detected in EORA, but not in YORA. Trans-complementing DQ heterodimer molecules, formed by DQA1 and DQB1 of the haplotypes with and without shared epitope alleles, might explain the higher genotype frequencies of "shared epitope /not shared epitope". Linear regression analyses showed the primary role of DQB104:01 allele for the age at onset of RA.This is the first report for the associations of DRB1 genotype with YORA or EORA in the Japanese population and the differential distribution of the genotypes was noted between these RA subsets. The involvement of DQ molecules for the age at onset of RA was suggested.


Asunto(s)
Edad de Inicio , Artritis Reumatoide , Cadenas HLA-DRB1/genética , Adulto , Anciano , Alelos , Artritis Reumatoide/diagnóstico , Artritis Reumatoide/epidemiología , Artritis Reumatoide/genética , Correlación de Datos , Femenino , Frecuencia de los Genes , Predisposición Genética a la Enfermedad , Humanos , Pruebas Inmunológicas/métodos , Japón/epidemiología , Masculino , Persona de Mediana Edad
11.
Zhonghua Nei Ke Za Zhi ; 58(12): 911-914, 2019 Dec 01.
Artículo en Chino | MEDLINE | ID: mdl-31775456

RESUMEN

The purpose of this study was to explore the role and mechanism of transient receptor potential M(2) (TRPM(2)) in antigen-induced arthritis (AIA) mice. Twelve C57BL/6 mice and 12 TRPM(2) knockout mice were divided into 4 groups, includingwild type control group, wild type AIA group, TRPM(2) knockout control group and TRPM(2) knockout AIA group, with 6 mice in each group. Methylated bovine serum albumin (mBSA) was used to establish AIA mouse model. The degree of joint swelling and inflammatory cell infiltration were recorded, as well as synovial hyperplasia of the knee joints. Real-time quantitative polymerase chain reaction (PCR) was used to detect the expression of interleukin (IL)-6, IL-8, chemokine ligand 6 (CXCL-6) and tumor necrosis factor alpha (TNFα) mRNA in synovial cells of knee joints. The results showed that compared with the wild-type AIA group, the TRPM(2) knockout AIA group had more significant synovial proliferation and inflammatory cell infiltration in the synovial tissue.The neutrophil and macrophage counts rather than monocytes in the knee joints of TRPM(2) knockout AIA group were higher than those in wild-type AIA mice. The expression of IL-6, IL-8 and CXCL-6 mRNA were significantly increased in the knock out mice. In summary, TRPM(2) may inhibit inflammatory cytokines such as IL-6 and IL-8 in knee joints of AIA mice by reducing the infiltration of neutrophils and macrophages, the refore alleviates the manifestations of knee arthritis.


Asunto(s)
Artritis Experimental/inmunología , Artritis Reumatoide/inmunología , Quimiocina CXCL6/inmunología , Interleucina-6/inmunología , Interleucina-8/inmunología , Albúmina Sérica Bovina/inmunología , Factor de Necrosis Tumoral alfa/inmunología , Animales , Antígenos/inmunología , Artritis Reumatoide/genética , Quimiocina CXCL6/genética , Interleucina-6/genética , Interleucina-8/genética , Articulación de la Rodilla/inmunología , Articulación de la Rodilla/patología , Ratones , Ratones Endogámicos C57BL , Reacción en Cadena en Tiempo Real de la Polimerasa , Membrana Sinovial , Factor de Necrosis Tumoral alfa/genética
13.
Genome Biol ; 20(1): 227, 2019 11 08.
Artículo en Inglés | MEDLINE | ID: mdl-31699133

RESUMEN

We present the software Condition-specific Regulatory Units Prediction (CRUP) to infer from epigenetic marks a list of regulatory units consisting of dynamically changing enhancers with their target genes. The workflow consists of a novel pre-trained enhancer predictor that can be reliably applied across cell types and species, solely based on histone modification ChIP-seq data. Enhancers are subsequently assigned to different conditions and correlated with gene expression to derive regulatory units. We thoroughly test and then apply CRUP to a rheumatoid arthritis model, identifying enhancer-gene pairs comprising known disease genes as well as new candidate genes.


Asunto(s)
Elementos de Facilitación Genéticos , Programas Informáticos , Animales , Artritis Experimental/genética , Artritis Reumatoide/genética , Código de Histonas , Ratones
14.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 41(5): 595-600, 2019 Oct 30.
Artículo en Chino | MEDLINE | ID: mdl-31699188

RESUMEN

Objective To explore the role of multidrug resistance gene-1(MDR1)gene in methotrexate(MTX)resistance in patients with rheumatoid arthritis(RA).Methods Fibroblast-like synoviocytes(FLS)from RA patients were infected with recombinant adenovirus Ad-EGFP-MDR1 in vitro to obtain MDR1 over-expressed RA FLS.The transcription level of MDR1 gene and the expression level of its coding product P-glycoprotein(P-gp) rotein were detected by real-time PCR and Western blot analysis.The efflux function was verified by rhodamine 123 efflux assay.The resistance to MTX was detected by MTT assay.Results RA FLS were infected with recombinant adenovirus Ad-EGFP-MDR1;72 hours later,the particles size in MDR1 over-expressed RA FLS increased,the cell volume became larger,and the growth rate decreased.The transcription level of MDR1(1.4325±0.3924 vs.0.0650±0.0070;t=6.035,P=0.004),the expression level of P-gp protein(1.8667±0.2857 vs. 0.9367±0.0551;t=5.536,P=0.005),and the ability of extracellular rhodamine 123(979.43±196.81 vs.1680.06±147.04;t=-4.940,P=0.008) in MDR1 over-expressed RA FLS were significantly higher than those of negative virus control RA-FLS,and the survival rate of MDR1 over-expressed RA FLS was significantly increased at each concentration of MTX(P<0.05).Conclusion The high expression of MDR1 can affect the efflux ability to MTX by up-regulating the expression of P-gp,thus enhancing the drug resistance to MTX in RA FLS.


Asunto(s)
Artritis Reumatoide/tratamiento farmacológico , Resistencia a Medicamentos , Fibroblastos/efectos de los fármacos , Metotrexato/farmacología , Subfamilia B de Transportador de Casetes de Unión a ATP/genética , Artritis Reumatoide/genética , Células Cultivadas , Humanos , Membrana Sinovial/citología
15.
Vestn Oftalmol ; 135(5. Vyp. 2): 192-198, 2019.
Artículo en Ruso | MEDLINE | ID: mdl-31691659

RESUMEN

Ophthalmologic manifestation of Sjogren's disease (SD) and rheumatoid arthritis (RA) is dry keratoconjunctivitis (dry eye disease; DED). PURPOSE: To study the relationship of polymorphic markers rs7947461 (C/T), rs915956 (C/T), rs4144331 (C/A) of the TRIM21 gene with the severity of DED in patients with RA and SD. MATERIAL AND METHODS: The study included 70 patients with RA (n=27) and SD (n=43). The control group consisted of volunteers without a history of RA or SD (n=35). Alleles of the polymorphic marker C660T rs7947461 of the TRIM21 gene were identified using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method; alleles of the polymorphic marker rs915956 (C/T) and rs4144331 (C/A) of the TRIM21 gene were identified by analyzing DNA melting curves. RESULTS: An association was found between the predisposing genotype (TT) of rs7947461 polymorphic marker and the risk of developing severe DED. The AA genotype of rs4144331 polymorphic marker was found only in severe DED (c2=7.74; OR=17.46, CI95%=1.96-318.38, p=0.02). CONCLUSION: An association was established between rs7947461 (rs660) and rs4144331 and the risk of developing severe DED.


Asunto(s)
Artritis Reumatoide , Queratoconjuntivitis , Ribonucleoproteínas/genética , Síndrome de Sjögren , Alelos , Artritis Reumatoide/genética , Predisposición Genética a la Enfermedad , Genotipo , Humanos , Queratoconjuntivitis/genética , Polimorfismo Genético , Síndrome de Sjögren/genética
16.
Mol Biol (Mosk) ; 53(5): 755-773, 2019.
Artículo en Ruso | MEDLINE | ID: mdl-31661476

RESUMEN

Dysregulated proinflammatory cytokine expression may result in the development of severe pathologies, such as rheumatoid arthritis, psoriasis, and neurodegenerative diseases. Transgenic mice and, in particular, those with controllable systemic overexpression of proinflammatory cytokines have recently become an essential instrument to study the molecular mechanisms underlying disease development. Importantly, many of the models are humanized by introducing a human cytokine gene, while leaving or removing the respective endogenous mouse gene. Humanized mice are especially valuable for biomedical research as they provide a relevant model to develop therapies based on blocking the pathogenic activity of a cytokine or to establish the functional significance of genome polymorphisms. The review discusses the available humanized mouse models with overexpression of key proinflammatory cytokines (TNF, IL-ip, and IL-6) and inflammatory cytokines with more specific functions (IL-8, IL-17, and IL-32) and their significance for basic and clinical research.


Asunto(s)
Citocinas/genética , Citocinas/metabolismo , Modelos Animales de Enfermedad , Mediadores de Inflamación/metabolismo , Regulación hacia Arriba , Animales , Artritis Reumatoide/genética , Citocinas/biosíntesis , Humanos , Ratones , Ratones Transgénicos , Enfermedades Neurodegenerativas/genética , Psoriasis/genética
17.
Ann Clin Lab Sci ; 49(5): 581-589, 2019 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-31611200

RESUMEN

OBJECTIVE: MiR-15 is involved in modulating many cellular processes that are normally destroyed in disease. The regulation of the cell cycle, angiogenesis, and responses to cellular stress are notable processes influenced by MiR-15. MiR-15 plays a critical role in the model of inflammation-related angiogenesis defects. However, its role in rheumatoid arthritis (RA) remains unclear. In this paper, we questioned the potential effects miR-15 has on RA. METHODS: Synthetic mimics of miR-15 were transfected into human RA fibroblast-like synoviocyte (FLS) cell line MH7A cells. Cell proliferation was detected by a Cell Counting Kit-8 (CCK-8) assay. Cell apoptosis was determined by flow cytometry. MH7A cells, stimulated by interleukin-1ß (IL-1ß), were transfected with miR-15 mimics and inhibitors. Western blot and quantitative real time (qRT)-PCR were used to detect gene and protein expressions. Bioinformatic software analysis and luciferase reporter assays were used to predict and validate miR-15 targets. RESULTS: We found that the miR-15 experienced down-regulated expression in MH7A cells, after they were stimulated with IL-1ß. MiR-15 inhibited inflammatory cell proliferation and promoted inflammatory cell apoptosis. BCL2L2 was identified as a target of miR-15 by luciferase assay. Additionally, NF-κB signaling pathway related proteins were inhibited by miR-15 overexpression. CONCLUSION: These findings contribute to an understanding of the impact that miR-15 possesses on inflammatory injury, and provide a basis for RA treatment.


Asunto(s)
Proteínas Reguladoras de la Apoptosis/genética , Artritis Reumatoide/genética , Eliminación de Gen , Regulación de la Expresión Génica , MicroARNs/genética , FN-kappa B/metabolismo , Transducción de Señal , Regiones no Traducidas 3'/genética , Apoptosis/efectos de los fármacos , Apoptosis/genética , Proteínas Reguladoras de la Apoptosis/metabolismo , Artritis Reumatoide/patología , Secuencia de Bases , Línea Celular , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/genética , Humanos , Interleucina-1beta/farmacología , MicroARNs/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/genética
18.
Scand J Rheumatol ; 48(5): 353-361, 2019 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-31631790

RESUMEN

Objective: To elucidate the roles of interleukin-6 (IL-6) and tumour necrosis factor-α (TNF-α) in cell cycle regulation and proliferation of rheumatoid arthritis fibroblast-like synovial cells (RA-FLSs). Methods: Under stimulation with IL-6/soluble interleukin-6 receptor (sIL-6R) and TNF-α, we examined the expression of cell cycle regulators [p16INK4a, p21Cip1, p27Kip1, cyclin-dependent kinase-4 (CDK4), CDK6, Cyclin D, Cyclin E, and retinoblastoma protein (pRB)] by quantitative polymerase chain reaction, Western blotting, and immunofluorescence staining. The expression of pRB, with or without 10% foetal bovine serum, was examined by Western blotting. DNA synthesis and cell viability were examined by the BrdU assay and WST-8 assay, respectively. After transfection with siRNA/p16INK4a, siRNA/p21Cip1, siRNA/p27Kip1, siRNA/CDK4, or siRNA/CDK6, RA-FLSs were successively stimulated with or without IL-6/sIL-6R or TNF-α to determine cell viability. Results: IL-6/sIL-6R significantly decreased the expression of p16INK4a, and increased p21Cip1, Cyclin E1, CYCLIN D, and pRB. TNF-α decreased the expression of CDK4, and significantly increased p27Kip1, CDK6, Cyclin E1/E2, CYCLIN D, CYCLIN E, pRB, and phosphorylated pRB (phospho-pRB). By immunofluorescence staining, CYCLIN D and phospho-pRB were simultaneously stained in the single cell. In serum-free culture, the expression of pRB was apparently decreased. DNA synthesis and cell viability were significantly increased by IL-6/sIL-6R and TNF-α. Silencing of CDK6 attenuated the cell viability induced by IL-6 and TNF-α. Conclusion: The results indicate that IL-6 and TNF-α interact with each other in regulating the cell cycle and accelerate the proliferation of RA-FLSs.


Asunto(s)
Artritis Reumatoide/genética , Regulación de la Expresión Génica , Interleucina-6/genética , Sinoviocitos/patología , Factor de Necrosis Tumoral alfa/genética , Artritis Reumatoide/metabolismo , Artritis Reumatoide/patología , Western Blotting , Ciclo Celular , Células Cultivadas , Ensayo de Inmunoadsorción Enzimática , Fibroblastos/metabolismo , Fibroblastos/patología , Humanos , Interleucina-6/biosíntesis , ARN/genética , Sinoviocitos/metabolismo , Factor de Necrosis Tumoral alfa/biosíntesis
19.
Molecules ; 24(20)2019 Oct 13.
Artículo en Inglés | MEDLINE | ID: mdl-31614911

RESUMEN

Combinations of IL-1ß and other proinflammatory cytokines reportedly promote the severity of arthritis. We aimed to investigate the effects of IL-1ß combined with IL-17A on cartilage degradation and synthesis in in vitro models. Cartilage explant degradation was determined using sulfated glycosaminoglycans (S-GAGs) levels, matrix metalloproteinase (MMP13) gene expression, uronic acid, and collagen contents. Cell morphology and accumulation of proteoglycans were evaluated using hematoxylin-eosin and safranin O staining, respectively. In the pellet culture model, expressions of cartilage-specific anabolic and catabolic genes were evaluated using real-time qRT-PCR. Early induction of MMP13 gene expression was found concomitantly with significant S-GAGs release. During the prolonged period, S-GAGs release was significantly elevated, while MMP-13 enzyme levels were persistently increased together with the reduction of the cartilaginous matrix molecules. The pellet culture showed anabolic gene downregulation, while expression of the proinflammatory cytokines, mediators, and MMP13 genes were elevated. After cytokine removal, these effects were restored to nearly basal levels. This study provides evidence that IL-1ß combined with IL-17A promoted chronic inflammatory arthritis by activating the catabolic processes accompanied with the suppression of cartilage anabolism. These suggest that further applications, which suppress inflammatory enhancers, especially IL-17A, should be considered as a target for arthritis research and therapy.


Asunto(s)
Artritis Reumatoide/genética , Interleucina-17/genética , Interleucina-1beta/genética , Metaloproteinasa 13 de la Matriz/genética , Animales , Artritis Reumatoide/metabolismo , Artritis Reumatoide/patología , Cartílago Articular/crecimiento & desarrollo , Cartílago Articular/metabolismo , Cartílago Articular/patología , Células Cultivadas , Condrocitos/metabolismo , Condrocitos/patología , Condrogénesis/genética , Regulación de la Expresión Génica , Glicosaminoglicanos/genética , Glicosaminoglicanos/metabolismo , Humanos , Inflamación/genética , Inflamación/patología , Metabolismo , Proteoglicanos/genética , Porcinos , Factor de Necrosis Tumoral alfa/genética
20.
Zhonghua Yi Xue Za Zhi ; 99(35): 2745-2749, 2019 Sep 17.
Artículo en Chino | MEDLINE | ID: mdl-31550796

RESUMEN

Objective: To investigatea cellular/molecular mechanism of the CD40/TRAF1 signalling pathway involved in Rheumatoid arthritis (RA). Methods: 16 patients with active RA and 9 patients with Fractures who underwent total knee or hip replacement in The First Affiliated Hospital of Soochow University were included in the study. Synovial tissues (ST) and serum were obtained from each patient. The CD40, TRAF1, NF-κB p65 were detected by ELISA and Immunohistochemistry in serum and tissue respectively. Real time-PCR (RT-PCR) was applied to measure NF-κB-related gene expression. Results: CD40 and TRAF1 positive area (%) in RA patients were 28.7±5.4, 34.3±4.8 respectively, which were significantly higher (P<0.05) than Fracture controls (21.2±9.5, 21.6±8.7 respectively). The expression of total NF-κB p65, and phospho-NF-κB p65 proteins, as well as NF-κB-related gene expression, including cytokines (TNFα, IL-6), chemokines (MCP-1),and adhesion molecules (ICAM-1) were significantly higher in the ST of RA patients compared to Fracture controls. Conclusion: It is thus possible that the CD40/TRAF1 pathway acted as a positive regulator through NF-κB activation and NF-κB-dependent proinflammatory genes in RA.


Asunto(s)
Artritis Reumatoide/genética , Antígenos CD40/metabolismo , Transducción de Señal , Factor 1 Asociado a Receptor de TNF/metabolismo , Factor de Transcripción ReIA/metabolismo , Antígenos CD40/genética , Células Cultivadas , Expresión Génica , Humanos , Membrana Sinovial , Factor 1 Asociado a Receptor de TNF/genética , Factor de Transcripción ReIA/genética
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA