scholarly journals AB0012 THE EFFECTS OF COMMON THERAPEUTICS ON AIRE METHYLATION STATUS AND THE LEVELS OF IL-16, IL-1Β, AND IFN-Γ IN LPS-INDUCED MACROPHAGE CELLS CAN SULPHASALAZINE’S INFLUENCE ON METHYLATION STATUS OF AIRE EXPLAIN ITS AUTOIMMUNE SIDE EFFECTS?

2021 ◽  
Vol 80 (Suppl 1) ◽  
pp. 1040.3-1041
Author(s):  
Z. B. Çelik ◽  
C. Gunaydin ◽  
A. K. Cengiz
Keyword(s):  
Rheumatoid Arthritis ◽  
Side Effects ◽  
Methylation Status ◽  
Treg Cells ◽  
Biological Properties ◽  
Drug Induced ◽  
Macrophage Cells ◽  
Drug Concentrations ◽  
Disease Modifying

Background:The autoimmune regulator gene (AIRE) has very important role in self-tolerance. Estrogen induced hypermethylation of AIRE promoter was put forward as one of the reasons for susceptibility to autoimmunity in females. [1] Sulphasalazine is a commonly used drug in rheumatology and is known to be responsible for various autoimmune side effects.Objectives:We aimed to investigate the effects of commonly used treatments on the methylation status of AIRE and levels of interleukin-16 (IL16), interleukin-1β (IL1β), and interferon-γ (IFNγ), in lipopolysaccharides (LPS)-induced RAW264.7 macrophage cells which mimic inflammatory state in rheumatoid arthritis in vitro.[2]Methods:RAW264.7 cells were stimulated by LPS (1μ/mL). Cell viability test was performed to determine drug concentrations. Following drug treatments, cell media was isolated for the determination of IL16, IL1β, and IFNγ levels by ELISA. Also, the cells detached by trypsin-EDTA were used for DNA isolation and bisulfite modification. Then, the promoter methylation status of AIRE was analyzed with methylation-specific PCR and agarose jel electrophoresis.Results:Our results demonstrated that the AIRE promoter is highly methylated in absence of any inflammatory stimulus.LPS treatment changed methylation status to unmethylated form. Leflunomide(LEF), sulfasalazine(SLZ) and methotrexate(MTX) significantly altered methylation status as the methylated (p=0.041,p<0.001,p<0.001, respectively). Surprisingly, SLZ treatment caused significantly more methylation in AIRE promoter than other drugs (p<0.001,Fig 1A). Analysis of IL16, IL1β and IFNγ levels demonstrated that LPS caused potent increase in all three cytokines (p<0.001, Fig 1B-D). LEF and SLZ significantly prevented LPS-induced increase in these cytokines (p=0.025, p<0.001 for IL-16, p<0.001 for IL-1β and IFNγ, Fig 1B-D). Although MTX exacerbated LPS-induced increase in IL16 levels, it inhibited LPS induced increase in IL1β and IFNγ levels (p<0.001, Fig 1B-D).Figure 1.PCR gel image. All data expressed as mean±SD. ***p<0.001 versus control, ###p<0.001, #p<0.05 versus LPS, +++p<0.001 versus sulfasalazine.Conclusion:Our results suggest that although LEF,SLZ and MTX potently suppresses LPS-induced inflammatory cytokines, they affect AIRE methylation differently.Changes in the methylation status of AIRE are important for autoimmunity. Relative increase in methylation of AIRE promoter by SLZ usage can be responsible for its autoimmune side effects, like drug induced lupus or hypersensitivity reactions, which are not common in MTX and LEF usage.IL16 is a key regulator of biological properties of CD4+T cells, it also regulates migration of CD4+CD25+Treg cells.[3] MTX treatment was shown to increase Treg cells in early rheumatoid arthritis patients. LEF and SLZ’s effect on Treg cells was shown to be different from that of MTX.[4] Our results demonstrated that MTX exacerbated LPS-induced increase in IL16 levels in contrary to LEF and SLZ. This difference may also be responsible for the different effect of these medications on Treg functions.The different effect of commonly used disease modifying drugs on IL16 levels and methylation of AIRE promoter is interesting and deserves attention.References:[1]Berrih-Aknin, Le Panse R, Dragin N. AIRE:a missing link to explain female susceptibility to autoimmune diseases. Ann.N.Y.Acad.Sci, 2018;1412(1):21-32.[2]Lin Y-Y, Jean Y-H, Lee H-P et al. Excavatolide B attenuates rheumatoid arthritis through the inhibition of osteoclastogenesis. Marine Drugs, 2017; 15(1):9.[3]Skundric DS, Cruikshank WW, Montgomery PC et al. Emerging role of IL-16 in cytokine-mediated regulation of multiple sclerosis. Cytokine, 2015;75(2):234-248.[4]Oh JS, Kim Y-G, Lee SG et al. The effect of various disease modifying anti-rheumatic drugs on suppressive function of CD4+CD25+ regulatory Tcells. Rhematol Int. 2013;33:381-388.Disclosure of Interests:None declared

scholarly journals Therapeutic Effect of Exogenous Treg Cells on Collagen-Induced Arthritis and Rheumatoid Arthritis

2019 ◽  
Author(s):  
Shutong Li ◽  
Hongxing Wang ◽  
Hui Wu ◽  
Guoqing Zhang ◽  
Xiaotian Chang
Keyword(s):  
Rheumatoid Arthritis ◽  
Flow Cytometry ◽  
Peripheral Blood ◽  
Synovial Fibroblast ◽  
Treg Cells ◽  
Cell Counting ◽  
Fibroblast Cells ◽  
Collagen Induced Arthritis ◽  
Gm Csf

Abstract Background Regulatory T (Treg) cells have anti-inflammatory and anti-autoimmune functions. The proportion and functions of Treg cells are perturbed in rheumatoid arthritis (RA) patients. Methods Human Treg cells were induced to amplify in vitro and cocultured with RA synovial fibroblast cells (RASFs). The proliferation and apoptosis of RASFs were determined by the cell counting kit-8 (CCK-8) assay and flow cytometry, respectively. Human Treg cells were also injected to collagen-induced arthritis (CIA) rats via the tail vein. Changes in lymphocyte subtypes and cytokines in the peripheral blood and spleen were observed by flow cytometry. Results After coculture with the Treg cells, the proliferation of RA synovial fibroblast cells decreased (p<0.01), and the rate of apoptosis increased (p=0.037). The human Treg cells were injected into the tail veins of collagen-induced arthritis (CIA) rats. The severity of the CIA was reduced (p<0.01) following the injection, the percentages of rat endogenous Treg cells in the peripheral blood and spleen increased significantly (p=0.007 and p<0.01, respectively), and the proportion of B cells decreased (p=0.031). The levels of interleukin IL-5 and IL-6 and the Th1/Th2 ratio in the peripheral blood were significantly decreased (p=0.013, 0.009 and 0.012, respectively). The number of NK cells and the levels of IL-4, IL-13, TNF-α, IFN-γ and GM-CSF in the peripheral blood and spleen did not change significantly. Conclusion These results suggest that exogenous Treg cells play a therapeutic role in RA and CIA. Treg cell treatment could serve as a therapy for RA.

Rheumatoid Arthritis: Strategies for Pharmacotherapy

1999 ◽  
Vol 12 (4) ◽  
pp. 271-281
Author(s):  
Bruce C. Carlstedt ◽  
Walter F. Stanaszek
Keyword(s):  
Rheumatoid Arthritis ◽  
Side Effects ◽  
Adverse Reactions ◽  
Initial Therapy ◽  
Disease Modifying Antirheumatic Drugs ◽  
Antirheumatic Drugs ◽  
Disease Modifying

Historically, treatment of rheumatoid arthritis (RA) has been conservative and symptom or patient-complaint oriented. Newer approaches use disease-modifying antirheumatic drugs (DMARDs), sometimes known as slow-acting antirheumatic drugs (SAARDs), to slow the progression of the disease earlier in its course. Some of these drugs may be used as initial therapy and some may be used in combinations to reduce the side effects or adverse reactions.

scholarly journals Anti-Inflammatory Effects of Formononetin 7-O-phosphate, a Novel Biorenovation Product, on LPS-Stimulated RAW 264.7 Macrophage Cells

Molecules ◽  
2019 ◽  
Vol 24 (21) ◽  
pp. 3910 ◽  
Author(s):  
Min-Seon Kim ◽  
Jin-Soo Park ◽  
You Chul Chung ◽  
Sungchan Jang ◽  
Chang-Gu Hyun ◽  
...  
Keyword(s):  
Biological Properties ◽  
In Vitro Assays ◽  
Raw 264.7 Cells ◽  
No Production ◽  
Raw 264.7 ◽  
Dependent Manner ◽  
Macrophage Cells ◽  
Anti Inflammatory ◽  
Reduced Toxicity

Biorenovation is a microbial enzyme-catalyzed structural modification of organic compounds with the potential benefits of reduced toxicity and improved biological properties relative to their precursor compounds. In this study, we synthesized a novel compound verified as formononetin 7-O-phosphate (FMP) from formononetin (FM) using microbial biotransformation. We further compared the anti-inflammatory properties of FMP to FM in lipopolysaccharide (LPS)-treated RAW264.7 macrophage cells. We observed that cell viabilities and inhibitory effects on LPS-induced nitric oxide (NO) production were greater in FMP-treated RAW 264.7 cells than in their FM-treated counterparts. In addition, FMP treatment suppressed the production of proinflammatory cytokines such as prostaglandin-E2 (PGE2), interleukin-6 (IL-6), and interleukin-1β (IL-1β) in a dose-dependent manner and concomitantly decreased the mRNA expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2). We also found that FMP exerted its anti-inflammatory effects through the downregulation of the extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and nuclear factor kappa B (NF-κB) signaling pathways. In conclusion, we generated a novel anti-inflammatory compound using biorenovation and demonstrated its efficacy in cell-based in vitro assays.

Adherence Rates and Associations with Nonadherence in Patients with Rheumatoid Arthritis Using Disease Modifying Antirheumatic Drugs

2009 ◽  
Vol 36 (10) ◽  
pp. 2164-2170 ◽  
Author(s):  
BART J.F. van den BEMT ◽  
FRANK H.J. van den HOOGEN ◽  
BART BENRAAD ◽  
YECHIEL A. HEKSTER ◽  
PIET L.C.M. van RIEL ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Risk Factors ◽  
Side Effects ◽  
Clinical Characteristics ◽  
Coping Styles ◽  
Self Report ◽  
Disease Modifying Antirheumatic Drugs ◽  
Antirheumatic Drugs ◽  
Potential Risk Factors ◽  
Disease Modifying

Objective.Nonadherence in patients with rheumatoid arthritis (RA) using disease modifying antirheumatic drugs (DMARD) may result in unnecessarily high levels of disease activity and function loss. The aim of this descriptive study was to assess adherence rates with self-report measures in a large random population, and to identify potential risk factors for nonadherence.Methods.A randomly selected sample of 228 patients with RA using DMARD was invited for a standardised interview. For each medicine, the patients were asked about adherence, consumption and perceived (side) effects. After the interview, the patients received self-report questionnaires to assess adherence [Compliance Questionnaire on Rheumatology (CQR) and the Medication Adherence Scale (MARS)], coping, beliefs about medicines, satisfaction about medicine information, and physical functioning. Subsequently, associations between adherence and demographics, clinical characteristics, and patient attitudes were examined.Results.Depending on the instrument used, 68% (CQR) and 60% (MARS) of the patients were adherent to DMARD. Nonadherence was not associated with demographic and clinical characteristics, satisfaction about information, medication concerns, and coping styles. The disease duration, the number of perceived side-effects, and beliefs about the necessity of the medicine were weakly associated with adherence.Conclusion.In this large study with a random RA population, 32%–40% of the patients did not adhere to their DMARD prescription. As none of the possible risk factors was strongly related to adherence, no general risk factor seems to be powerful enough as a possible screening tool or target for adherence-improving interventions. This implies that nonadherence barriers should be assessed on an individual basis.

scholarly journals Immediate Effect of Baricitinib on Arthritis and Biological Disease-Modifying Antirheumatic Drug-Induced Psoriasis-Like Skin Lesions in Two Patients with Rheumatoid Arthritis

2021 ◽  
Vol 2021 ◽  
pp. 1-4
Author(s):  
Yoshifumi Tada ◽  
Nobuyuki Ono ◽  
Syuichi Koarada
Keyword(s):  
Rheumatoid Arthritis ◽  
Adverse Events ◽  
Certolizumab Pegol ◽  
Skin Lesions ◽  
Disease Modifying Antirheumatic Drugs ◽  
Antirheumatic Drugs ◽  
Drug Induced ◽  
Beneficial Effects ◽  
Disease Modifying ◽  
Disease Modifying Antirheumatic Drug

Biological disease-modifying antirheumatic drugs (bDMARDs) are very effective for treating rheumatoid arthritis (RA). However, they sometimes induce adverse events such as psoriasis-like skin lesions. We describe psoriasis-like skin lesions that developed simultaneously with an RA flare in patient 1 during treatment with abatacept and in patient 2 soon after starting certolizumab pegol. The skin lesions persisted in patient 2 despite stopping certolizumab. Baricitinib was initiated because of RA flare and resulted in immediate beneficial effects on arthritis as well as skin lesions. The RA went into remission in both patients, and the psoriasis-like skin lesions disappeared within four weeks (patient 1) and three months (patient 2).

Chiral Forms of 4-(2',4'-Difluorobiphenyl-4-yl)-2-methyl-4-oxobutanoic Acid (Flobufen) and Its Metabolite.Synthesis and Basic Biological Properties

1997 ◽  
Vol 62 (3) ◽  
pp. 498-509 ◽  
Author(s):  
Miroslav Kuchař ◽  
Marie Poppová ◽  
Antonín Jandera ◽  
Vladimíra Panajotovová ◽  
Hana Zůnová ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Biological Activities ◽  
Biological Properties ◽  
Optical Isomers ◽  
Clinical Testing ◽  
Chemical Parameters ◽  
Physico Chemical ◽  
Oxobutanoic Acid

4-(2',4'-Difluorobiphenyl-4-yl)-2-methyl-4-oxobutanoic acid (1, flobufen) is subjected to clinical testing in the treatment of rheumatoid arthritis. Owing to the occurrence of a centre of chirality, the compound exists in two enantiomers, and its major human metabolite, viz. 4-(2',4'-difluorobiphenyl-4-yl)-4-hydroxy-2-methylbutanoic acid isolated in the lactone form (2), possesses two chiral centres, making possible the existence of four stereoisomers. All of the optical isomers of the substances 1 and 2 were prepared. For flobufen (1), the racemate was separated into the stereoisomers by using the salts 3 with R-(+)- or S-(-)-1-phenylethylamine. The pairs of stereoisomers of 2, obtained by reduction of R-(+)-flobufen or the S-(-)-enantiomer, were separated by column chromatography. The physico-chemical parameters of the optical isomers were determined and some biological activities were evaluated in both in vitro and in vivo models.

scholarly journals PREPARATION AND EVALUATION OF SULFASALAZINE LOADED SODIUM ALGINATE MICROBEADS FOR SUSTAINED DELIVERY

2016 ◽  
pp. 72
Author(s):  
Krishna Sailaja A ◽  
Jyothika Mattam
Keyword(s):  
Rheumatoid Arthritis ◽  
Side Effects ◽  
Drug Release ◽  
Zeta Potential ◽  
Sodium Alginate ◽  
Process Parameters ◽  
Megaloblastic Anemia ◽  
Entrapment Efficiency ◽  
Sustained Delivery

<p>ABSTRACT<br />Objective: The main objective of the work was to prepare and evaluate sulfasalazine loaded sodium alginate microbeads for sustained delivery for<br />the treatment of inflammatory bowel disease and rheumatoid arthritis. Sulfasalazine has crystalluria, thrombocytopenia, and megaloblastic anemia<br />as side effects, so to reduce side effect microbeads were prepared.<br />Methods: The sulfasalazine microbeads were prepared by inotropic gelation method by optimizing process parameters such as concentration of<br />calcium chloride, agitation speed, and time of agitation. The concentration of polymer sodium alginate was varied.<br />Result: Among the five formulations, the best formulation was considered by comparing process parameters such as the entrapment efficiency, drug<br />content, in vitro drug release studies, scanning electron microscope analysis, and zeta potential.<br />Conclusion: On comparison, B3 formulation was considered as best formulation with a mean particle size ranging from 40.9 to 244 µm, drug content<br />of 94.7%, entrapment efficiency of 87.7%, and the drug release was found to be 97.1% for 12 hrs and followed zero order kinetics and non-Fickian<br />diffusional pathway, with a zeta potential value of −56.8 mV indicating higher stability.<br />Keywords: Inotropic gelation method, Sodium alginate, Microbeads, Rheumatoid arthritis, Side effects.</p>

scholarly journals Tafenoquine is a Promising Drug Candidate for the Treatment of Babesiosis

2021 ◽  
Author(s):  
Mingming Liu ◽  
Shengwei Ji ◽  
Daisuke Kondoh ◽  
Eloiza May Galon ◽  
Jixu Li ◽  
...  
Keyword(s):  
Side Effects ◽  
Complete Blood Count ◽  
Survival Rates ◽  
Vital Signs ◽  
Repeated Administration ◽  
Farm Animals ◽  
Drug Candidate ◽  
Radical Cure ◽  
Drug Induced

Due to drug resistance, commonly used anti-Babesia drugs have limited efficacy against babesiosis, and inflict severe side effects. Tafenoquine (TAF) was approved by the U.S. Food and Drug Administration in 2018 for the radical cure of Plasmodium vivax infection and for malaria prophylaxis. Here, we evaluated the efficacy of TAF for the treatment of Babesia infection and elucidated the suspected mechanisms of TAF activity against Babesia parasites. Parasitemia and survival rates of B. rodhaini–infected BALB/c and SCID mice were used to explore the role of the immune response in Babesia infection after TAF treatment. Parasitemia, survival rates, body weight, vital signs, complete blood count and blood biochemistry of B. gibsoni–infected splenectomized dogs were determined to evaluate the anti-Babesia activity and side effects of TAF. Then, to understand the mechanism of TAF activity, hydrogen peroxide was used as an oxidizer for short-term B. rodhaini incubation in vitro, and the expression levels of antioxidant enzymes were confirmed using B. microti–infected mice by qRT-PCR. Acute B. rodhaini and B. gibsoni infections were rapidly eliminated with TAF administration. Repeated administration of TAF or a combination therapy with other antibabesial agents is still needed to avoid a potentially fatal recurrence for immunocompromised hosts. Caution about hyperkalemia should be taken during TAF treatment for Babesia infection. TAF possesses a babesicidal effect that may be related to drug-induced oxidative stress. Considering the lower frequency of glucose-6-phosphate dehydrogenase deficiency in animals than in humans, TAF use on Babesia-infected farm animals and pets is eagerly anticipated.

scholarly journals Expression and Purification of an Active Form of the Mycobacterium leprae DNA Gyrase and Its Inhibition by Quinolones

2007 ◽  
Vol 51 (5) ◽  
pp. 1643-1648 ◽  
Author(s):  
Stéphanie Matrat ◽  
Stéphanie Petrella ◽  
Emmanuelle Cambau ◽  
Wladimir Sougakoff ◽  
Vincent Jarlier ◽  
...  
Keyword(s):  
Dna Cleavage ◽  
Active Form ◽  
Mycobacterium Leprae ◽  
Dna Gyrase ◽  
Dna Supercoiling ◽  
Drug Induced ◽  
Vitro Assay ◽  
Drug Concentrations

ABSTRACT Mycobacterium leprae, the causative agent of leprosy, is noncultivable in vitro; therefore, evaluation of antibiotic activity against M. leprae relies mainly upon the mouse footpad system, which requires at least 12 months before the results become available. We have developed an in vitro assay for studying the activities of quinolones against the DNA gyrase of M. leprae. We overexpressed in Escherichia coli the M. leprae GyrA and GyrB subunits separately as His-tagged proteins by using a pET plasmid carrying the gyrA and gyrB genes. The soluble 97.5-kDa GyrA and 74.5-kDa GyrB subunits were purified by nickel chelate chromatography and were reconstituted as an enzyme with DNA supercoiling activity. Based on the drug concentrations that inhibited DNA supercoiling by 50% or that induced DNA cleavage by 25%, the 13 quinolones tested clustered into three groups. Analysis of the quinolone structure-activity relationship demonstrates that the most active quinolones against M. leprae DNA gyrase share the following structural features: a substituted carbon at position 8, a cyclopropyl substituent at N-1, a fluorine at C-6, and a substituent ring at C-7. We conclude that the assays based on DNA supercoiling inhibition and drug-induced DNA cleavage on purified M. leprae DNA gyrase are rapid, efficient, and safe methods for the screening of quinolone derivatives with potential in vivo activities against M. leprae.

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