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1.
Biochem Pharmacol ; 195: 114869, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-34896056

RESUMO

NFκB plays a key role in inflammation and skeletal disorders. Previously, we reported that pharmacological inhibition of NFκB at the level of TRAF6 suppressed RANKL, CD40L and IL1ß-induced osteoclastogenesis and attenuated cancer-induced bone disease. TNFα is also known to regulate TRAF6/NFκB signalling, however the anti-inflammatory and osteoprotective effects associated with inhibition of the TNFα/TRAF6/NFκB axis have not been investigated. Here, we show that in vitro and ex vivo exposure to the verified small-molecule inhibitor of TRAF6, 6877002 prevented TNFα-induced NFκB activation, osteoclastogenesis and calvarial osteolysis, but it had no effects on TNFα-induced apoptosis or growth inhibition in osteoblasts. Additionally, 6877002 disrupted T-cells support for osteoclast formation and synoviocyte motility, without affecting the viability of osteoblasts in the presence of T-cells derived factors. Using the collagen-induced arthritis model, we show that oral and intraperitoneal administration of 6877002 in mice reduced joint inflammation and arthritis score. Unexpectedly, no difference in trabecular and cortical bone parameters were detected between vehicle and 6877002 treated mice, indicating lack of osteoprotection by 6877002 in the arthritis model described. Using two independent rodent models of osteolysis, we confirmed that 6877002 had no effect on trabecular and cortical bone loss in both osteoporotic rats or RANKL- treated mice. In contrast, the classic anti-osteolytic alendronate offered complete osteoprotection in RANKL- treated mice. In conclusion, TRAF6 inhibitors may be of value in the management of the inflammatory component of bone disorders, but may not offer protection against local or systemic bone loss, unless combined with anti-resorptive therapy such as bisphosphonates.


Assuntos
Anti-Inflamatórios/farmacologia , Antígenos CD40/antagonistas & inibidores , Osteólise/prevenção & controle , Fator 6 Associado a Receptor de TNF/antagonistas & inibidores , Animais , Anti-Inflamatórios/química , Artrite Experimental/metabolismo , Artrite Experimental/prevenção & controle , Antígenos CD40/metabolismo , Linhagem Celular Tumoral , Humanos , Células Jurkat , Masculino , Camundongos , Camundongos Endogâmicos C3H , Camundongos Endogâmicos DBA , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , Osteoclastos/citologia , Osteoclastos/efeitos dos fármacos , Osteoclastos/metabolismo , Osteogênese/efeitos dos fármacos , Osteólise/metabolismo , Células RAW 264.7 , Roedores/metabolismo , Fator 6 Associado a Receptor de TNF/metabolismo , Fator de Necrose Tumoral alfa/farmacologia
2.
Phytomedicine ; 94: 153841, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34752968

RESUMO

BACKGROUND: 7-Hydroxycoumarin (7-HC) as a coumarin compound is widely found in Chinese herbs and exhibits diverse biological activities. Promoting cell apoptosis of fibroblast-like synoviocytes (FLS) is a meaningful strategy for rheumatoid arthritis (RA). Though the protective effect of 7-HC on RA experimental models has been reported, the specific mechanisms, especially the possible relationships of this effect to regulating FLS proliferation and apoptosis, still need clarification. PURPOSE: This study clarified the therapeutic effects of 7-HC on collagen-induced arthritis (CIA) in rats and explored the underlying mechanisms. METHODS: In vivo, 7-HC (15, 30 or 60 mg/kg) was intraperitoneally given to CIA rats, and its therapeutic effect and anti-inflammatory activity were evaluated. Ki67 immunohistochemistry, TUNEL assay and synovial proteins detection were conducted. In vitro, after treating with 7-HC (20, 40 or 80 µM) in TNF-α-stimulated RA FLS (MH7A cell line), cell proliferation and apoptosis were examined. The involvement of Wnt/ß-catenin pathway was checked in vivo and in vitro. RESULTS: 7-HC attenuated the severity of rat CIA, evidenced by the reduction of paw swelling, arthritis index, joint damage, collagen type II antibody serum level, and IL-1ß, IL-6, TNF-α production in serum and synovium. Particularly, 7-HC in vivo had anti-proliferative and pro-apoptotic effects on CIA rat synovial cells, indicated by reduced synovial Ki67 expression, raised synovial apoptosis index, decreased Bcl-2 protein level and increased level of Bax and cleaved caspase 3 protein. Further, 7-HC in vitro suppressed proliferation and promoted apoptosis of TNF-α-stimulated MH7A cells by regulating the mitochondrial pathway. Mechanistically, 7-HC treatment inhibited Wnt/ß-catenin pathway, suggested by the reduction of pathway-related proteins (e.g. Wnt1, LRP6, p-GSK-3ß (Ser9), ß-catenin, cyclin D1 and c-Myc), the recovery of GSK-3ß activity and the inhibition of ß-catenin nuclear translocation. As expected, combined use of lithium chloride, an activator of Wnt/ß-catenin signaling, reversed the anti-proliferative and pro-apoptotic effects of 7-HC in vitro. CONCLUSION: 7-HC relieved the severity of rat CIA by inhibiting cell proliferation and inducing apoptosis of rheumatoid FLS via inhibition of Wnt/ß-catenin pathway.


Assuntos
Artrite Experimental , Sinoviócitos , Animais , Apoptose , Artrite Experimental/tratamento farmacológico , Proliferação de Células , Células Cultivadas , Fibroblastos , Glicogênio Sintase Quinase 3 beta , Ratos , Membrana Sinovial , Umbeliferonas/farmacologia , Via de Sinalização Wnt
3.
Zhongguo Zhong Yao Za Zhi ; 46(22): 5895-5901, 2021 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-34951180

RESUMO

Sophorae Flavescentis Radix, the root of Sophora flavescens Ait., has been widely applied in the medical field due to its anti-inflammatory, analgesic, bacteriostatic, antiviral, antitumor, and other pharmacological effects. The present study investigated the anti-rheumatoid arthritis effect of oxymatrine(OMT), the active component of Sophorae Flavescentis Radix by observing its effect on the function of B lymphocytes in collagen-induced arthritis(CIA) mice through the Toll-like receptor 9(TLR9)/myeloid differentiation factor 88(MyD88)/signal transducer and activator of transcription 3(STAT3) pathway. The CIA model in DBA/1 J mice was induced by bovine type Ⅱ collagen and complete Freund's adjuvant(CFA). Fifteen days after the primary immunization, mice were treated with OMT for 30 days by intraperitoneal injection. Paw swelling and arthritis index(AI) score were evaluated every 3 days. Joint histopathologic changes were observed by HE staining. Magnetic-activated cell sorting(MACS) was used to isolate B lymphocytes from the spleen of CIA mice spleen. The serum expression level of interleukin(IL)-21 was examined by the enzyme-linked immunosorbent assay(ELISA). The expression of TLR9, STAT3, p-STAT3, and IL-21 in B lymphocytes was detected by Western blot. The mRNA expression of TLR9, STAT3, and IL-21 in B lymphocytes was detected by real-time fluorescence-based quantitative PCR(qRT-PCR). The results showed that OMT could significantly alleviate the paw swelling, decrease the AI score, relieve synovial inflammatory cell infiltration and hyperplasia, reduce the level of inflammatory cytokines, and inhibit the expression of TLR9, STAT3, p-STAT3, and IL-21 of B lymphocytes in CIA mice. Therefore, OMT may alleviate rheumatoid arthritis by regulating TLR9/MyD88/STAT3 pathway in B lymphocytes, providing a valuable reference for the application of OMT in the clinical treatment of rheumatoid arthritis.


Assuntos
Alcaloides , Artrite Experimental , Animais , Artrite Experimental/tratamento farmacológico , Artrite Experimental/genética , Bovinos , Citocinas , Camundongos , Camundongos Endogâmicos DBA , Quinolizinas
4.
Cell Mol Life Sci ; 79(1): 3, 2021 Dec 16.
Artigo em Inglês | MEDLINE | ID: mdl-34913099

RESUMO

Autoimmune arthritis is characterized by impaired regulatory T (Treg) cell migration into inflamed joint tissue and by dysregulation of the balance between Treg cells and Th17 cells. Interleukin-6 (IL-6) is known to contribute to this dysregulation, but the molecular mechanisms behind impaired Treg cell migration remain largely unknown. In this study, we assessed dynamic changes in membrane-bound IL-6 receptor (IL6R) expression levels on Th17 cells by flow cytometry during the development of collagen-induced arthritis (CIA). In a next step, bioinformatics analysis based on proteomics was performed to evaluate potential pathways affected by altered IL-6R signaling in autoimmune arthritis. Our analysis shows that membrane-bound IL-6R is upregulated on Th17 cells and is inversely correlated with IL-6 serum levels in experimental autoimmune arthritis. Moreover, IL-6R expression is significantly increased on Th17 cells from untreated patients with rheumatoid arthritis (RA). Interestingly, CD4+ T cells from CIA mice and RA patients show reduced phosphorylation of vasodilator-stimulated phosphoprotein (VASP). Bioinformatics analysis based on proteomics of CD4+ T cells with low or high phosphorylation levels of VASP revealed that integrin signaling and related pathways are significantly enriched in cells with low phosphorylation of VASP. Specific inhibition of p-VASP reduces the migratory function of Treg cells but has no influence on effector CD4+ T cells. Importantly, IL-6R blockade restores the phosphorylation level of VASP, thereby improving the migratory function of Treg cells from RA patients. Thus, our results establish a link between IL6R signaling and phosphorylation of VASP, which controls Treg cell migration in autoimmune arthritis.


Assuntos
Artrite Reumatoide/patologia , Moléculas de Adesão Celular/metabolismo , Proteínas dos Microfilamentos/metabolismo , Fosfoproteínas/metabolismo , Receptores de Interleucina-6/metabolismo , Linfócitos T Reguladores/metabolismo , Células Th17/metabolismo , Animais , Artrite Experimental/induzido quimicamente , Artrite Experimental/metabolismo , Artrite Experimental/patologia , Artrite Reumatoide/metabolismo , Linfócitos T CD8-Positivos/citologia , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/metabolismo , Movimento Celular , Humanos , Interleucina-6/sangue , Camundongos , Camundongos Endogâmicos DBA , Fosforilação , Linfócitos T Reguladores/citologia , Células Th17/citologia , Regulação para Cima
5.
J Environ Pathol Toxicol Oncol ; 40(4): 43-52, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34936299

RESUMO

The disease progression of rheumatoid arthritis (RA) is closely related to the disorder of amino acids metabolism. This study aimed to clarify the changes in the categories, quantities and metabolic pathways of amino acids associated with disease progression in adjuvant-induced arthritis (AIA) rats, and to evaluate the application value of amino acids metabolic profiling in the diagnosis of RA. A total of 20 rats were randomized into a control group and AIA model group. Thirty-three days after modeling, the synovial tissues of left ankle joints were collected for histopathological examination. Also, untargeted metabolomics based on UPLC-LTQ/Orbitrap MS was used to identify the potential amino acids and analyze the changes of their metabolic pathways. Histopathological observation showed that the synovial tissues of AIA rats exhibited hyperplastic and inflammatory lesions. Through multivariate statistical analysis, a total of 12 amino acids were identified and considered to show metabolic changes in AIA rats. Compared with the control group, the concentrations of glutamate, arginine, methionine were increased in the AIA model group; while the concentrations of proline, valine, tyrosine, phenylalanine, leucine, glycine, tryptophan, histidine, threonine were decreased. Metabolic pathway analysis showed that the selected amino acids had important physiological functions in immune and inflammatory response. The results suggested that amino acids could be employed as biomarkers closely related to RA, and the analysis of amino acids metabolic profiling also exhibited potential application value in the diagnosis, disease progression monitoring and therapy of RA.


Assuntos
Aminoácidos/metabolismo , Artrite Experimental/metabolismo , Artrite Reumatoide/metabolismo , Progressão da Doença , Metabolômica/métodos , Animais , Masculino , Ratos , Ratos Sprague-Dawley
6.
Zhen Ci Yan Jiu ; 46(12): 1016-22, 2021 Dec 25.
Artigo em Chinês | MEDLINE | ID: mdl-34970878

RESUMO

OBJECTIVE: To observe the effect of acupuncture of "Yinlingquan"(SP9) and "Sanyinjiao"(SP6) on expression of phosphatidylinositol-3 kinase/protein kinase B/mammalian target protein of rapamycin (PI3K/Akt/mTOR) signaling in adjuvant arthritis (AA) rats, so as to explore its mechanism underlying improvement of AA. METHODS: Forty-eight male Wistar rats were randomly divided into normal control, AA model, acupuncture and medication (tripterygium wilfordii) groups, with 12 rats in each group. The AA model was established by putting the rats in a windy, cold and wet environment for 12 h, once every day for 21 days and injection of Freund's complete adjuvant (CFA) into the sole of the right hindlimb on the 21st day. Manual acupuncture stimulation was applied at SP9 and SP6 for 30 min/time, once a day for 21 days. Rats of the medication group received gavage of tripterygium wilfordii tablets solution (8 mg/kg), once a day for 21 days, and those of the normal control group and model group received gavage of the same amount of normal saline, once a day for 21 days. The degree of joint swelling and arthritis index (AI) were detected 1 day before modeling, 3 days after modeling, and 21 days after the treatment. Twenty-four hours after the last treatment, the contents of serum cytokines interleukin (IL)-17, IL-6 and tumor necrosis factor (TNF)-α were detected by enzyme-linked immunosorbent assay (ELISA); and changes of synovial ultrastructure were observed under electron microscope. Western blot was used to detect the expression levels of PI3K, Akt, phosphorylated protein kinase B (p-Akt), phosphorylated mammalian rapamycin target protein (p-mTOR), mTOR, microtubule associated protein 1 light chain 3B (LC3-Ⅱ) and mammalian atg6 homologous protein (Beclin-1) in the synovial membrane tissue. RESULTS: Compared with the normal control group, the joint swel-ling degree and AI, contents of serum TNF-α, IL-6 and IL-17, and the expression levels of PI3K, Akt, p-Akt, mTOR and p-mTOR in the synovium were increased in the model group (P<0.05), while the expression levels of LC3-Ⅱ and Beclin-1 proteins in the synovium were significantly decreased (P<0.05). Compared with the model group, the joint swelling degree and AI, contents of serum TNF-α, IL-6 and IL-17, and the expression levels of PI3K, Akt, p-Akt, mTOR and p-mTOR were significantly decreased in the acupuncture and medication groups (P<0.05), while the expresion levels of LC3-Ⅱ and Beclin-1 proteins were significantly increased (P<0.05). Comparison between the two treatment groups showed that the therapeutic effects of acupuncture were obviously weaker than those of medication in down-regulating TNF-α and IL-6, Akt, p-Akt and mTOR levels (P<0.05) and in up-regulating Beclin-1 expression (P<0.05). Outcomes of electron microscope displayed widened nuclear membrane space, some fractured mitochondrial cristae with vacuoles, expanded rough endoplasmic reticulum, reduction of autophagosomes in the cytoplasm and ruptured synovial cell membrane in the model group, and increase of autophagosomes, deformed organelles in the acupuncture and medication groups. CONCLUSION: Acupuncture can relieve the inflammatory reactions and joint synovial injury of the affected joint in AA rats which may be associated with its effects in inhibiting PI3K/Akt/mTOR signaling and increasing the auto-phagy level of synovial cells.


Assuntos
Terapia por Acupuntura , Artrite Experimental , Animais , Artrite Experimental/genética , Artrite Experimental/terapia , Autofagia , Masculino , Fosfatidilinositol 3-Quinases/genética , Proteínas Proto-Oncogênicas c-akt/genética , Ratos , Ratos Sprague-Dawley , Ratos Wistar , Serina-Treonina Quinases TOR/genética
7.
Int J Mol Sci ; 22(24)2021 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-34948139

RESUMO

Vitamin D plays a crucial role in regulation of the immune response. However, treatment of autoimmune diseases with 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] doses sufficient to be effective is prohibitive due to its calcemic and toxic effects. We use the collagen-induced arthritis (CIA) model to analyze the efficacy of the noncalcemic analog of vitamin D, 20S-hydroxyvitamin D3 [20S(OH)D3], as well as 1,25(OH)2D3, to attenuate arthritis and explore a potential mechanism of action. Mice fed a diet deficient in vitamin D developed a more severe arthritis characterized by enhanced secretion of T cell inflammatory cytokines, compared to mice fed a normal diet. The T cell inflammatory cytokines were effectively suppressed, however, by culture of the cells with 20S(OH)D3. Interestingly, one of the consequences of culture with 1,25(OH)2D3 or 20S(OH)D3, was upregulation of the natural inhibitory receptor leukocyte associated immunoglobulin-like receptor-1 (LAIR-1 or CD305). Polyclonal antibodies which activate LAIR-1 were also capable of attenuating arthritis. Moreover, oral therapy with active forms of vitamin D suppressed arthritis in LAIR-1 sufficient DR1 mice, but were ineffective in LAIR-1-/- deficient mice. Taken together, these data show that the effect of vitamin D on inflammation is at least, in part, mediated by LAIR-1 and that non-calcemic 20S(OH)D3 may be a promising therapeutic agent for the treatment of autoimmune diseases such as Rheumatoid Arthritis.


Assuntos
Artrite Experimental/metabolismo , Calcifediol/análogos & derivados , Calcitriol/farmacologia , Receptores Imunológicos/biossíntese , Linfócitos T/metabolismo , Regulação para Cima/efeitos dos fármacos , Animais , Artrite Experimental/tratamento farmacológico , Artrite Experimental/genética , Artrite Experimental/patologia , Calcifediol/farmacologia , Camundongos , Camundongos Knockout , Receptores Imunológicos/genética , Linfócitos T/patologia
8.
Phytomedicine ; 93: 153801, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34758437

RESUMO

BACKGROUND: Rheumatoid arthritis (RA) is a common autoimmune disease, for which no economical and safe target drug treatment is available. Chikusetsusaponin Ⅳa (CS-IVa), an active compound in Panax japonicus C.A. Mey, has a good anti-inflammatory effect, but whether this compound can serve as a targeted drug for RA and the corresponding therapeutic mechanism remain unclear. PURPOSE: To investigate the anti-inflammatory and bone-protecting effects of CS-IVa on RA and the possible corresponding mechanisms of action. METHODS: Biomarkers and underlying pathological mechanisms were examined by performing a bioinformatics analysis of RA synovial gene expression data profiles, and the feasibility of CS-IVa treatment for RA was predicted using molecular docking and molecular dynamics simulation techniques. Histomorphological and molecular biology techniques were used to verify the feasibility and molecular mechanism of CS-IVa treatment for RA in vivo using a collagen-induced arthritis (CIA) model. RESULTS: CS-IVa alleviated symptoms and reduced the immune organ index, arthritis index, hind paw thickness, and number of swollen joints in the foot for CIA mice. Bioinformatics analysis suggested that interferon-gamma (IFN-γ), interleukin-1 ß (IL-1ß), and the Janus kinase/signal transduction and activator of transcription (JAK/STAT) pathway played important roles in the pathogenesis of RA. The results of molecular docking and molecular dynamics simulations showed that CS-IVa bound effectively to IFN-γ and IL-1ß and that the combined pose has good stability and flexibility. The histomorphological results showed that CS-IVa reduced joint histopathology scores, OARSI scores, and TRAP-positive cell counts. Molecular biology analysis indicated that CS-IVa reduced the concentration of inflammatory factors in the peripheral serum of CIA mice and suppressed the mRNA expression of these factors in the spleen in a dose-dependent manner. The protein expression level of the JAK/STAT pathway was also inhibited by CS-IVa. CONCLUSION: The results of the current study demonstrate a novel inhibitory effect of CS-IVa on inflammation and bone destruction in CIA mice, and the mechanism may be related to the JAK/STAT signaling pathway, which provides new insights into the development of CS-IVa as a therapeutic agent for RA.


Assuntos
Artrite Experimental , Artrite Reumatoide , Animais , Anti-Inflamatórios/farmacologia , Artrite Experimental/tratamento farmacológico , Artrite Reumatoide/tratamento farmacológico , Janus Quinases/metabolismo , Camundongos , Simulação de Acoplamento Molecular , Ácido Oleanólico/análogos & derivados , Saponinas , Transdução de Sinais
9.
Zhongguo Ying Yong Sheng Li Xue Za Zhi ; 37(5): 449-453, 2021 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-34816649

RESUMO

Objective: An animal model of collagen-induced arthritis (CIA) was used to investigate the effects of norepinephrine (NE) and α1-adrenoreceptor (α1-AR) on Treg cells in CIA. Methods: Thirty-two male DBA/1 mice were randomly divided into control group and CIA model group. CIA was prepared by intradermal injection of collagen type II (CII, 100 µl) at the tail base of DBA/1 mice. On the 41th day following primary immunization, co-expression of CD4+T and α1-AR in mouse spleens was observed. Protein expressions of α1-AR in the ankle joints and the spleens of mice were measured by Western blot analysis. The CD4+ T cells were isolated from the mouse spleen tissues in CIA mice and treated with NE or α1-AR agonist phenylephrine. Percentage of Treg cells in mouse CD4+ T cells of CIA mice was determined by flow cytometry. Expressions of α1-AR, transforming growth factor-ß (TGF-ß) and IL-10 in CD4+T cells of CIA mice were assessed by Western blot. Results: Co-expression of CD4 and α1-AR was observed in spleens of both intact and CIA mice. Compared with intace mice, α1-AR expressions in the ankle joints and spleens were down-regulated in CIA mice. NE increased the function of Treg cells in CD4+ T cells of CIA mice compared with that of nothing-treated CD4+T cells of CIA mice. Moreover, the α1-AR agonist phenylephrine increased the Treg cell function in CD4+ T cells of CIA mice relative to that of nothing-treated CD4+T cells of CIA mice. Conclusion: Activating α1-AR on CD4+T cells of CIA mice enhances Treg cell function,facilitating a shift of CD4+T cells toward Treg polarization.


Assuntos
Artrite Experimental , Animais , Colágeno Tipo II , Masculino , Camundongos , Camundongos Endogâmicos DBA , Transdução de Sinais , Linfócitos T Reguladores
10.
Arthritis Res Ther ; 23(1): 265, 2021 10 25.
Artigo em Inglês | MEDLINE | ID: mdl-34696809

RESUMO

BACKGROUND: The development and optimization of therapies for rheumatoid arthritis (RA) is currently hindered by a lack of methods for early non-invasive monitoring of treatment response. Annexin A2, an inflammation-associated protein whose presence and phosphorylation levels are upregulated in RA, represents a potential molecular target for tracking RA treatment response. METHODS: LS301, a near-infrared dye-peptide conjugate that selectively targets tyrosine 23-phosphorylated annexin A2 (pANXA2), was evaluated for its utility in monitoring disease progression, remission, and early response to drug treatment in mouse models of RA by fluorescence imaging. The intraarticular distribution and localization of LS301 relative to pANXA2 was determined by histological and immunohistochemical methods. RESULTS: In mouse models of spontaneous and serum transfer-induced inflammatory arthritis, intravenously administered LS301 showed selective accumulation in regions of joint pathology including paws, ankles, and knees with positive correlation between fluorescent signal and disease severity by clinical scoring. Whole-body near-infrared imaging with LS301 allowed tracking of spontaneous disease remission and the therapeutic response after dexamethasone treatment. Histological analysis showed preferential accumulation of LS301 within the chondrocytes and articular cartilage in arthritic mice, and colocalization was observed between LS301 and pANXA2 in the joint tissue. CONCLUSIONS: We demonstrate that fluorescence imaging with LS301 can be used to monitor the progression, remission, and early response to drug treatment in mouse models of RA. Given the ease of detecting LS301 with portable optical imaging devices, the agent may become a useful early treatment response reporter for arthritis diagnosis and drug evaluation.


Assuntos
Anexina A2 , Artrite Experimental , Artrite Reumatoide , Animais , Artrite Experimental/diagnóstico por imagem , Artrite Experimental/tratamento farmacológico , Artrite Reumatoide/diagnóstico por imagem , Artrite Reumatoide/tratamento farmacológico , Condrócitos , Camundongos , Imagem Óptica , Tirosina
11.
In Vivo ; 35(6): 3245-3251, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34697155

RESUMO

BACKGROUND/AIM: Using a rat model of collagen-induced arthritis (CIA), we evaluated the therapeutic effects of HM71224 (BTKi), as well as the drug-drug interactions in combined therapy with methotrexate (MTX) based on both drugs' pharmacological role in immune regulation and antiinflammation. MATERIALS AND METHODS: Arthritis in rats was induced using type II collagen and incomplete Freund's adjuvant. The therapeutic effects of HM71224 (alone or in combination with MTX) were evaluated by arthritis score, paw volume, body weight, and histopathological examination (H&E and Safranin-O staining). The drug-drug interactions between HM71224 and MTX were investigated by measuring plasma, liver enzyme and creatinine levels and blood cell counts. RESULTS: HM71224 reduced the clinical signs of arthritis, paw volume, and body weight loss in CIA rats. ED50 and ED90 were 1.0 and 2.5 mg/kg, respectively. HM71224 combined with MTX decreased the arthritis score, bone erosion, synovitis, and cartilage degradation without apparent interaction. CONCLUSION: The combination of HM71224 and MTX improved the therapeutic effect with no drug-drug interactions in RA.


Assuntos
Artrite Experimental , Metotrexato , Animais , Anti-Inflamatórios/uso terapêutico , Artrite Experimental/tratamento farmacológico , Quimioterapia Combinada , Metotrexato/farmacologia , Metotrexato/uso terapêutico , Inibidores de Proteínas Quinases/uso terapêutico , Ratos
12.
Arthritis Res Ther ; 23(1): 273, 2021 10 29.
Artigo em Inglês | MEDLINE | ID: mdl-34715926

RESUMO

BACKGROUND: Central nervous system (CNS)-mediated symptoms, such as fatigue, depression, and hyperalgesia, are common complications among patients with rheumatoid arthritis (RA). However, it remains unclear how the peripheral pathology of RA spreads to the brain. Accumulated evidence showing an association between serum cytokine levels and aberrant CNS function suggests that humoral factors participate in this mechanism. In contrast to the well-known early responses of microglia (CNS-resident immune cells) in the area postrema [AP; a brain region lacking a blood-brain barrier (BBB)] to experimental inflammation, microglial alterations in the AP during chronic inflammation like RA remain unclear. Therefore, to determine whether microglia in the AP can react to persistent autoimmune-arthritis conditions, we analyzed these cells in a mouse model of collagen-induced arthritis (CIA). METHODS: Microglial number and morphology were analyzed in the AP of CIA and control mice (administered Freund's adjuvant or saline). Immunostaining for ionized calcium-binding adaptor molecule-1 was performed at various disease phases: "pre-onset" [post-immunization day (PID) 21], "establishment" (PID 35), and "chronic" (PID 56 and 84). Quantitative analyses of microglial number and morphology were performed, with principal component analysis used to classify microglia. Interleukin-1ß (IL-1ß) mRNA expression was analyzed by multiple fluorescent in situ hybridization and real-time polymerase chain reaction. Behavioral changes were assessed by sucrose preference test. RESULTS: Microglia in the AP significantly increased in density and exhibited changes in morphology during the establishment and chronic phases, but not the pre-onset phase. Non-subjective clustering classification of cell morphology (CIA, 1,256 cells; saline, 852 cells) showed that the proportion of highly activated microglia increased in the CIA group during establishment and chronic phases. Moreover, the density of IL-1ß-positive microglia, a hallmark of functional activation, was increased in the AP. Sucrose preferences in CIA mice negatively correlated with IL-1ß expression in brain regions containing the AP. CONCLUSIONS: Our findings demonstrate that microglia in the AP can sustain their activated state during persistent autoimmune arthritis, which suggests that chronic inflammation, such as RA, may affect microglia in brain regions lacking a BBB and have various neural consequences.


Assuntos
Artrite Experimental , Artrite Reumatoide , Animais , Área Postrema , Humanos , Hibridização in Situ Fluorescente , Camundongos , Microglia
13.
Stem Cell Res Ther ; 12(1): 535, 2021 10 09.
Artigo em Inglês | MEDLINE | ID: mdl-34627365

RESUMO

BACKGROUND: Tumor necrosis factor (TNF)-α inhibitors represented by Etanercept (a fusion protein containing soluble TNF receptor II (sTNFRII) and the Fc segment of human IgG1) play a pivotal role in Rheumatoid arthritis (RA) treatment. However, long-term use increases the risk of infection and tumors for their systemic inhibition of TNF-α, which disrupts the regular physiological function of this molecular. Mesenchymal stem cells (MSCs)-based delivery system provides new options for RA treatment with their "homing" and immune-regulation capacities, whereas inflammatory environment (especially TNF-α) is not conducive to MSCs' therapeutic effects by inducing apoptosis/autophagy. Here, we constructed a strain of sTNFRII-Fc-expressing MSCs (sTNFRII-MSC), aiming to offset the deficiency of those two interventions. METHODS: Constructed sTNFRII-Fc lentiviral vector was used to infect human umbilical cord-derived MSCs, and sTNFRII-MSC stable cell line was generated by monoclonal cultivation. In vitro and vivo characteristics of sTNFRII-MSC were assessed by coculture assay and an acute inflammatory model in NOD/SCID mice. The sTNFRII-MSC were transplanted into CIA model, pathological and immunological indicators were detected to evaluate the therapeutic effects of sTNFRII-MSC. The distribution of sTNFRII-MSC was determined by immunofluorescence assay. Apoptosis and autophagy were analyzed by flow cytometry, western blot and immunofluorescence. RESULTS: sTNFRII-Fc secreted by sTNFRII-MSC present biological activity both in vitro and vivo. sTNFRII-MSC transplantation effectively alleviates mice collagen-induced arthritis (CIA) via migrating to affected area, protecting articular cartilage destruction, modulating immune balance and sTNFRII-MSC showed prolonged internal retention via resisting apoptosis/autophagy induced by TNF-α. CONCLUSION: sTNFRII-Fc modification protects MSCs against apoptosis/autophagy induced by TNF-α, in addition to releasing sTNFRII-Fc neutralizing TNF-α to block relevant immune-inflammation cascade, and thus exert better therapeutic effects in alleviating inflammatory arthritis.


Assuntos
Artrite Experimental , Transplante de Células-Tronco Mesenquimais , Animais , Apoptose , Autofagia , Humanos , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Fator de Necrose Tumoral alfa/genética
14.
Zhongguo Zhen Jiu ; 41(10): 1119-25, 2021 Oct 12.
Artigo em Chinês | MEDLINE | ID: mdl-34628745

RESUMO

OBJECTIVE: To observe the effect of moxibustion at "Zusanli" (ST 36) and "Shenshu" (BL 23) on inflammatory factors and intestinal flora in the rats with adjuvant arthritis. METHODS: A total of 36 Wistar rats were randomized into a normal group, a model group and a moxibustion group, 12 rats in each one. In the model group and the moxibustion group, the adjuvant arthritis model was established by a compound method, including the environmental factors, i.e. wind, cold and damp, and Freund's complete adjuvant. In the moxibustion group, moxibustion intervention was exerted at "Zusanli" (ST 36) and "Shenshu" (BL 23), for 20 min at each acupoint, once daily, consecutively for 21 days. The paw swelling degree and arthritis index (AI) score were observed before and after intervention in the rats of each group. Using real-time fluorescence quantitative PCR method (real-time PCR) and Western blot method, the mRNA and protein expressions of inflammatory factors of colon tissue, i.e. interleukin (IL) 1ß, tumor necrosis factor-α (TNF-α), IL-6, were detected after intervention in the rats of each group. The intestinal flora was detected with 16SrRNA sequencing technology after intervention in the rats of each group. RESULTS: Compared with the normal group, the paw swelling degree and AI score were increased in the rats of the model group (P<0.05); after intervention, compared with the model group, the paw swelling degree and AI score were reduced in the rats of the moxibustion group (P<0.05). Compared with the normal group, the expressions of IL-1ß, TNF-α and IL-6 mRNA, as well as proteins were increased in the colon tissue of the rats in the model group (P<0.05); compared with the model group, the expressions of IL-1ß, TNF-α and IL-6 mRNA, as well as proteins were reduced in the colon tissue of the rats in the moxibustion group (P<0.05). Compared with the normal group, OTUs count was reduced in the rats of the model group (P<0.05); and compared with the model group, OTUs count was increased in the rats of the moxibustion group (P<0.05). Compared with the normal group, Simpson index was increased, Chao1 and Ace were reduced in the rats of the model group (P<0.05); while, compared with the model group, Chao1 and Ace were increased in the rats of the moxibustion group (P<0.05). Compared with the normal group, the relative abundance of uncategorized Clostridium, Lactobacillus, Prevotella, uncategorized Porphyromonadaceae and uncategorized Prevotella was increased (P<0.05), and the relative abundance of uncategorized Spironella was reduced in the model group (P<0.05). While, compared with the model group, the relative abundance of uncategorized Clostridium, Lactobacillus, uncategorized Prevotella was reduced in the moxibustion group (P<0.05) and that of uncategorized Spironella was increased (P<0.05). CONCLUSION: Moxibustion at "Zusanli" (ST 36) and "Shenshu" (BL 23) relieves the joint symptoms of adjuvant arthritis rats and inhibits the expressions of inflammatory factors, which is probably related to the regulation of the structure of intestinal flora.


Assuntos
Artrite Experimental , Artrite Reumatoide , Microbioma Gastrointestinal , Moxibustão , Animais , Artrite Experimental/genética , Artrite Experimental/terapia , Ratos , Ratos Wistar
15.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 37(10): 865-871, 2021 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-34670662

RESUMO

Objective To investigate the effects of lncRNA-H19 on fibroblast-like synovial cells(FLS) and collagen induced arthritis (CIA) mice. Methods With FLS isolated and cultured from patients with rheumatoid arthritis (RA), adenovirus, lentivirus and siRNA were used to up-regulate and down-regulate the expression of H19 in RA FLS. BrdU, TranswellTM assay and cell scratch assay were employed to evaluate the proliferation, invasion and migration of RA FLS, respectively. Mice with CIA were locally injected with LV-sh-H19. The progression of CIA was observed by paw thickness, clinical arthritic index, and histologic analysis. Results The expression of H19 was closely associated with the proliferation, invasion and migration of FLS cells and knockdown of H19 significantly ameliorated the progression of CIA in mice. Conclusion H19 is involved in synovial inflammation and progression in CIA mice by promoting the activation of FLS.


Assuntos
Artrite Experimental , RNA Longo não Codificante/metabolismo , Animais , Artrite Experimental/genética , Proliferação de Células , Células Cultivadas , Fibroblastos , Humanos , Inflamação/genética , Camundongos , RNA Longo não Codificante/genética
16.
Zhen Ci Yan Jiu ; 46(8): 649-55, 2021 Aug 25.
Artigo em Chinês | MEDLINE | ID: mdl-34472749

RESUMO

OBJECTIVE: To observe the effect of electroacupuncture (EA) of "Zusanli"(ST36) and "Sanyinjiao"(SP6) on serum TNF-α, IL-1ß, and IL-6 and expression of synovial matrimetalloproteinases (MMPs) and articular morphology in collagen-induced arthritis (CIA) rats, so as to explore its mechanisms underlying relief of arthritis. METHODS: Thirty male SD rats were randomly divided into normal control, CIA model and EA groups (n=10 rats per group). The arthritis model was induced by multi-point intradermal injection of bovine type Ⅱ collagen emulsion. EA (2 Hz/100 Hz, 1 mA) was applied to bilateral ST36 and SP6 for 30 min, once a day for 28 days. The hind-limb paw volume was measured and the arthritis index (AI) score given according to the swelling degree, rigidity and deformity of the ankle joint (0-4 points). After EA intervention, the morphological damage of the affected ankle joints was revealed by H.E. staining, safranin O-fast green staining, and tartrate-resistant acid phosphatase (TRAP) staining, separately. The levels of serum TNF-α, IL-1ß, and IL-6 were measured by ELISA, and the expression levels of MMP-1, MMP-3, MMP-13, and receptor activator of nuclear factor Kappa B ligand (RANKL) in the synovial tissue were detected by Western blot. RESULTS: Compared with the normal control group, the paw volume, AI score, TRAP-revealed number of osteoclasts, contents of serum TNF-α, IL-1ß and IL-6, and expression levels of MMP-1, MMP-3, MMP-13 and RANKL proteins were significantly increased in the model group (P<0.01, P<0.05). Following the intervention, the paw volume, AI score, number of osteoclasts, contents of serum TNF- α, IL-1ß and IL-6, and expression levels of MMP-1, MMP-3, MMP-13 and RANKL proteins were significantly decreased in the EA group (P<0.05, P<0.01) in contrast to the model group. H.E. and safranin O-fast green staining showed rough articular cartilage surface with thinned cartilage layer, obvious hyperplasia of the synovial tissue with many inflammatory cells, and serious damage and degradation of the cartilage matrix in the model group, these situations were relatively milder in the EA group. CONCLUSION: EA of ST36 and SP6 can reduce the articular damage in collagen-induced arthritis rats, which is associated with its function in reducing inflammatory response and down-regulating the expression of synovial MMP-1, MMP-3, MMP-13 and RANKL proteins.


Assuntos
Artrite Experimental , Eletroacupuntura , Animais , Artrite Experimental/genética , Artrite Experimental/terapia , Bovinos , Inflamação/genética , Inflamação/terapia , Masculino , Ratos , Ratos Sprague-Dawley , Membrana Sinovial
17.
Biomed Res Int ; 2021: 5574282, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34497850

RESUMO

Programmed cell death 1 ligand (PD-L1) and its receptor (PD-1) are key molecules for immunoregulation and immunotherapy. PD-L1 binding PD-1 is an effective way to regulate T or B cell immunity in autoimmune diseases such as rheumatoid arthritis (RA). In our study, we overexpressed PD-L1 by constructing a recombinant of PD-L1-lentiviral vector, which was subsequently used to transfect mouse bone marrow mesenchymal stem cells (MBMMSCs) and significantly suppressed the development of collagen-induced arthritis (CIA) in DBA/1j mice. In addition, PD-L1-transfected MBMMSCs (PD-L1-MBMMSCs) ameliorated joint damage, reduced proinflammatory cytokine expression, and inhibited T and B cell activation. Furthermore, PD-L1-MBMMSCs decreased the number of dendritic cells and increased the numbers of regulatory T cells and regulatory B cells in joints of CIA mice. In conclusion, our results provided a potential therapeutic strategy for RA treatment with PD-L1-MBMMSC-targeted therapy.


Assuntos
Artrite Experimental/terapia , Artrite Reumatoide/terapia , Antígeno B7-H1/administração & dosagem , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/citologia , Linfócitos T Reguladores/imunologia , Animais , Artrite Experimental/imunologia , Artrite Experimental/patologia , Artrite Reumatoide/imunologia , Artrite Reumatoide/patologia , Células Cultivadas , Modelos Animais de Doenças , Ativação Linfocitária , Masculino , Células-Tronco Mesenquimais/imunologia , Células-Tronco Mesenquimais/metabolismo , Camundongos , Camundongos Endogâmicos DBA
18.
Sci Rep ; 11(1): 17971, 2021 09 09.
Artigo em Inglês | MEDLINE | ID: mdl-34504248

RESUMO

Rheumatoid arthritis (RA) is an autoimmune disease associated with advanced joint dysfunction. Madhuca indica J. F. Gmel, from the family Sapotaceae, is an Indian medicinal plant reported to have an array of pharmacological properties. The aim of present investigation was to determine the anti-arthritic potential of an isolated phytoconstituent from methanolic leaf extract of Madhuca indica (MI-ALC) against FCA-induced experimental arthritis. Polyarthritis was induced in female rats (strain: Wistar) via an intradermal injection of FCA (0.1 mL) into the tail. Polyarthritis developed after 32 days of FCA administration. Then rats were treated orally with an isolated phytoconstituent from MI-ALC at doses of 5, 10, and 20 mg/kg. Findings suggested that High-Performance Thin-Layer Chromatography, Fourier-Transform Infrared Spectroscopy, and Liquid Chromatography-Mass Spectrometry spectral analyses of the phytoconstituent isolated from MI-ALC confirmed the structure as 3,5,7,3',4'-Pentahydroxy flavone (i.e., QTN). Treatment with QTN (10 and 20 mg/kg) showed significant (p < 0.05) inhibition of increased joint diameter, paw volume, paw withdrawal threshold, and latency. The elevated synovial oxidative stress (Superoxide dismutase, reduced glutathione, and malondialdehyde) and protein levels of Tumor necrosis factor-α (TNF-α) and Interleukin (ILs) were markedly (p < 0.05) reduced by QTN. It also effectively (p < 0.05) ameliorated cyclooxygenase-2 (COX-2), Nuclear factor of kappa light polypeptide gene enhancer in B cells (NF-kß) and its inhibitor-α (Ikßα), and ATP-activated P2 purinergic receptors (P2X7) protein expressions as determined by western blot analysis. In conclusion, QTN ameliorates FCA-induced hyperalgesia through modulation of elevated inflammatory release (NF-kß, Ikßα, P2X7, and COX-2), oxido-nitrosative stress, and pro-inflammatory cytokines (ILs and TNF-α) in experimental rats.


Assuntos
Antirreumáticos/administração & dosagem , Artrite Experimental/tratamento farmacológico , Artrite Reumatoide/tratamento farmacológico , Flavonoides/administração & dosagem , Madhuca/química , Fitoterapia/métodos , Extratos Vegetais/administração & dosagem , Plantas Medicinais/química , Adjuvantes Imunológicos/efeitos adversos , Administração Oral , Animais , Antirreumáticos/química , Antirreumáticos/isolamento & purificação , Artrite Experimental/induzido quimicamente , Artrite Experimental/metabolismo , Artrite Reumatoide/induzido quimicamente , Artrite Reumatoide/metabolismo , Ciclo-Oxigenase 2/metabolismo , Citocinas/metabolismo , Feminino , Flavonoides/química , Flavonoides/isolamento & purificação , Adjuvante de Freund/efeitos adversos , Hiperalgesia/induzido quimicamente , Hiperalgesia/tratamento farmacológico , Hiperalgesia/metabolismo , Estrutura Molecular , NF-kappa B/metabolismo , Estresse Nitrosativo/efeitos dos fármacos , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Ratos , Ratos Wistar , Transdução de Sinais/efeitos dos fármacos , Resultado do Tratamento
19.
J Mech Behav Biomed Mater ; 124: 104719, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34481308

RESUMO

Rheumatoid arthritis (RA) is a systemic polyarticular arthritis that primarily affects the small joints but also causes bone erosion in large joints. None of the currently existing treatment approaches is curable. In this study, the effects of human gingiva-derived mesenchymal stem cells (GMSCs) on collagen-induced arthritis (CIA) mice are examined by experimentally assessing the microstructure and mechanical behaviors of tibia. Bone morphology and mineral density of mouse tibiae were assessed using micro-X-ray computed tomography (micro-CT). Compression testing was performed on mouse tibia to access its stiffness. The deformation and strain localized inside proximal tibia were mapped using mechanical testing coupled with micro-CT and digital volume correlation of micro-CT images. The results show that CIA disease caused bone erosion in epiphyseal cortical bone, which manifested into the adjacent epiphyseal trabecular bone, and also affected the metaphyseal cortical bone. CIA disease also weakened the load-bearing function of proximal tibia. GMSC treatment interfered with the progress of CIA, attenuated the bone erosion in epiphyseal and metaphyseal trabecular bone and resulted in improved load-bearing function of proximal tibia. GMSCs provide a promising potential treatment of autoimmune arthritis.


Assuntos
Artrite Experimental , Células-Tronco Mesenquimais , Animais , Colágeno , Gengiva , Camundongos , Tíbia/diagnóstico por imagem
20.
J Immunol ; 207(7): 1755-1762, 2021 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-34470853

RESUMO

Conformation-specific Ags are ideal targets for mAb-based immunotherapy. Here, we demonstrate that the monomeric form of C-reactive protein (mCRP) is a specific therapeutic target for arthritis and nephritis in a murine model. Screening of >1800 anti-mCRP mAb clones identified 3C as a clone recognizing the monomeric, but not polymeric, form of CRP. The anti-mCRP mAb suppressed leukocyte infiltration in thioglycollate-induced peritonitis, attenuated rheumatoid arthritis symptoms in collagen Ab-induced arthritis model mice, and attenuated lupus nephritis symptoms in MRL/Mp-lpr/lpr lupus-prone model mice. These data suggest that the anti-mCRP mAb 3C has therapeutic potential against rheumatoid arthritis and lupus nephritis.


Assuntos
Anticorpos Monoclonais/uso terapêutico , Artrite Experimental/imunologia , Artrite Reumatoide/imunologia , Proteína C-Reativa/imunologia , Imunoterapia/métodos , Nefrite Lúpica/imunologia , Peritonite/imunologia , Pleura/metabolismo , Animais , Anticorpos Monoclonais/metabolismo , Artrite Reumatoide/terapia , Modelos Animais de Doenças , Humanos , Nefrite Lúpica/terapia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos DBA , Camundongos Endogâmicos MRL lpr , Peritonite/terapia , Ligação Proteica , Conformação Proteica , Isoformas de Proteínas , Toracentese
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