scholarly journals Pinolenic acid Mediated Anti-inflammatory and Immunometabolic Effects in Peripheral blood-derived Monocytes from patients with Rheumatoid Arthritis is Associated with Modulation of miRNA expression

2022 ◽  
Author(s):  
Rabaa Takala ◽  
Dipak Ramji ◽  
Robert Andrews ◽  
You Zhou ◽  
Mustafa Farhat ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Mitochondrial Dysfunction ◽  
Pathway Analysis ◽  
Peripheral Blood ◽  
Interleukin 1 ◽  
Pyruvate Dehydrogenase Kinase ◽  
Metabolic Effects ◽  
Pinolenic Acid ◽  
Tnf Α ◽  
Anti Inflammatory

Abstract Objectives: Pinolenic acid (PNLA), an omega-6 polyunsaturated fatty acid from pine nuts, has anti-inflammatory and anti-atherogenic effects. We aimed to investigate the actions of PNLA on activated purified monocytes from peripheral blood of patients with rheumatoid arthritis (RA).Methods: Flow cytometry was used to assess the intracellular expression of TNF-α, IL-6, IL-1β, and IL-8 in purified monocytes from patients with RA after lipopolysaccharide (LPS) stimulation with/without PNLA pre-treatment. The whole genomic transcriptomic (WGT) profile of PNLA-treated, and LPS-activated monocytes from patients with active RA was investigated by RNA-sequencing.Results: PNLA reduced percentage of monocytes expressing the cytokines TNF-α by 23% (p=0.048), IL-6 by 25% (p=0.011), IL-1β by 23% (p=0.050) and IL-8 by 19% (p=0.066). Canonical pathway analysis showed that PNLA inhibited oxidative phosphorylation (p= 9.14E-09) and mitochondrial dysfunction (p=4.18E-08), while the sirtuin (SIRTs) signalling pathway was activated (p=8.89E-06). Pathway analysis predicted upstream activation of peroxisome proliferator-activated receptors (PPARs), sirtuin3, and let7miRNA, which are anti-inflammatory and antioxidative. In contrast, DAP3, LIF and STAT3, which are involved in TNF-α, and IL-6 signal transduction, were inhibited. Many miRNAs were modulated by PNLA suggesting potential post-transcriptional regulation of metabolic and immune response that has not been described previously. Multiple miRNAs target pyruvate dehydrogenase kinase-4 (PDK4), single-immunoglobulin interleukin-1 receptor-related molecule (SIGIRR), mitochondrially encoded ATP synthase membrane subunit 6 (MT-ATP6) and acetyl-CoA acyltransferase 2 (ACAA2); genes implicated in cell metabolism, inflammation, and mitochondrial dysfunction.Conclusion: PNLA has anti-inflammatory and immune-metabolic effects on monocytes that are pathogenic in RA and atherosclerosis. Dietary PNLA supplementation may regulate key miRNAs that are involved in mitochondrial, metabolic, and inflammatory pathways.

scholarly journals POS0346 THE THERAPEUTIC EFFECTS OF OMEGA-6 PUFA (PINOLENIC ACID) ON THE TRANSCRIPTOMIC PROFILE OF ACTIVATED PERIPHERAL BLOOD MONONUCLEAR CELLS ISOLATED FROM HEALTHY CONTROLS AND RHEUMATOID ARTHRITIS PATIENTS

2021 ◽  
Vol 80 (Suppl 1) ◽  
pp. 402-403
Author(s):  
R. Takala ◽  
D. Ramji ◽  
R. Andrews ◽  
Y. Zhou ◽  
J. Burston ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Rheumatic Diseases ◽  
Pathway Analysis ◽  
Healthy Controls ◽  
Pinolenic Acid ◽  
Eular Recommendations ◽  
Tnf Α ◽  
Omega 6 ◽  
Disease Modifying ◽  
Anti Inflammatory

Background:Globally, Rheumatoid arthritis (RA) is the most common chronic inflammatory arthritis. EULAR recommends patient education and disease modifying anti-rheumatic drugs including advanced targeted therapies for management1 2. Many patients seek advice on a dietary intervention that may improve symptoms. Foods high in omega-3 polyunsaturated fatty acids (PUFAs) is one of the most common recommended based on their anti-inflammatory properties3 4. Previously we showed pinolenic acid (PNLA), an omega-6 (PUFAs) found in pine nuts, reduced lipid uptake, endocytosis and migration of monocyte in vitro. PNLA also reduced IL-6, TNF-α, IL-1β and PGE2 produced by lipopolysaccharide (LPS) activated PBMCs from patients with RA and healthy controls (HCs).Objectives:We hypothesis that PNLA inhibits key inflammatory processes in synovities as in diagram.1. We investigated the transcriptomic profile of PNLA treatment on LPS activated PBMCs isolated from HCs and RA patients.Methods:Adult RA patients (n=6) were recruited from the Rheumatology Department of University Hospital of Wales together with 6 HCs. Blood was collected after taking signed informed consent. PBMCs were isolated by Ficoll gradient centrifugation and pre-treated with 25 μM PNLA or vehicle and then activated with 100 ng/ml LPS. RNA was extracted followed by library construction and sequencing for whole genomic transcriptome. DEGs were analysed using DESeq2 and pathway analysis was performed using Ingenuity pathway analysis. The study was approved by the Research Ethics Committee for Wales (reference no. 12/WA/0045).Results:DEGs were selected with at least differential genes using at least 1.2* fold changes and adjusted p-value of <0.05. PNLA significantly upregulates the expression of PDK4, PAI-1 (SERPINE1), FBP1, and NDRG2. These genes have important roles in metabolic pathways. Pathway analysis predicted upstream activation of nuclear receptors peroxisome proliferator-activated receptor (PPARs) involved in the anti-inflammatory process, and inhibition of NF-κB and STAT1, the major transcription factors of the pro-inflammatory cytokines TNF-α, IL-1, IL-6 and IFN-γ.Conclusion:PNLA has significant effects on the regulation of metabolic and inflammatory pathways in PBMCs from RA patients and HCs. Based on these results we demonstrate that PNLA may have beneficial anti-inflammatory effects in patients with RAReferences:[1]Agca R, Heslinga SC, Rollefstad S, Heslinga M, McInnes. IB, Peters MJL, Kvien, TK., Dougados, M, Radner, H, Atzeni, F. and Primdahl, J. 2017. EULAR recommendations for cardiovascular disease risk management in patients with rheumatoid arthritis and other forms of inflammatory joint disorders: 2015/2016 update. Annals of the rheumatic diseases, 76(1), pp.17-28.[2]Smolen JS, Landewé RBM, Bijlsma JWJ, et al. 2019. EULAR recommendations for the management of rheumatoid arthritis with synthetic and biological disease-modifying antirheumatic drugs: updateAnnals of the Rheumatic Diseases 2020; 79:685-699.[3]Chehade L, Jaafar ZA, El Masri D, Zmerly H, Kreidieh D, Tannir H, Itani L, and El Ghoch, M. 2019. Lifestyle Modification in Rheumatoid Arthritis: Dietary and Physical Activity Recommendations Based on Evidence. Current rheumatology reviews, 15(3), pp.209-214.[4]Marchand NE, Chiu Y-H, Yoshida K, Malspeis S, Sparks JA, Costenbader K, et al. Threshold Level for Long-term Healthy Diet Adherence to Reduce the Risk of Rheumatoid Arthritis Among Women in a Prospective Cohort Using a Marginal Structural Model Approach.:25.Diagram 1.Disclosure of Interests:None declared

scholarly journals P201 Anti-inflammatory and immunologic actions of pinolenic acid in rheumatoid arthritis

Rheumatology ◽  
2020 ◽  
Vol 59 (Supplement_2) ◽  
Author(s):  
Rabaa Takala ◽  
Dipak Ramji ◽  
Ernest Choy
Keyword(s):  
Rheumatoid Arthritis ◽  
Flow Cytometry ◽  
Mononuclear Cells ◽  
Vehicle Control ◽  
Boyden Chamber ◽  
Pinolenic Acid ◽  
Human Macrophages ◽  
Monocyte Migration ◽  
Tnf Α ◽  
Anti Inflammatory

Abstract Background Rheumatoid arthritis (RA) is a common inflammatory arthritis. Although advanced targeted therapies have improved prognosis, many patients seek advice on dietary intervention that may improve symptoms. Pinolenic acid (PNLA) is a polyunsaturated fatty acid found in pine nuts. We investigated the anti-inflammatory effects of 25-100 μM PNLA on cell line, primary culture, and peripheral blood mononuclear cells (PBMCs) from patients with RA and healthy controls (HCs). Methods 1- Migration using modified Boyden Chambers: THP-1 monocytes incubated with vehicle or PNLA were added to the apical compartment of a modified Boyden chamber. The migration of the cells through inserts of 8 μm pore size in response to the chemokine monocyte chemoattractant protein-1 (MCP-1) added to basolateral (bottom) chamber was determined. 2- Macropinocytosis using Lucifer yellow (LY): THP-1 and primary human macrophages were pre-incubated with PNLA or vehicle control followed by LY. After incubation, cells were removed, fixed and assessed by flow cytometry. 3- Lipid uptake using Dil-oxidised low-density lipoprotein (Dil-oxLDL): THP-1 and primary macrophages were pre-incubated with PNLA or vehicle control followed by Dil-oxLDL. After incubation, cells were removed, fixed and assessed by flow cytometry. 4- Cytokines release by lipopolysaccharide (LPS) stimulated PBMCs: PBMCs were isolated from blood obtained from RA patients aged ≥18 years and HCs. Monocytes were purified and cultured with PNLA or vehicle control. Cells were stimulated with LPS. IL-6, TNF-α, IL-1β and prostaglandin E2 (PGE2) in the supernatant were assessed by ELISAs. For macrophages, monocytes were left to grow and differentiate over 10 days, the differentiated macrophages were treated with PNLA or vehicle and activated with LPS and assayed in identical conditions for monocytes. Results PNLA at all concentrations reduced THP-1 monocytes migration by average of 55% (p &lt; 0.001) when compared with vehicle controls. Macropinocytosis of THP-1 macrophages and human macrophages were reduced by almost 50% (p &lt; 0.001) and 45% (p &lt; 0.001) respectively by PNLA. Similarly, Dil-oxLDL uptake by THP-1 macrophages and primary macrophages were reduced by 40% (p &lt; 0.01) and 25% (p &lt; 0.05) respectively by 25 μM PNLA. Release of IL-6 and TNF-α by LPS stimulated monocytes in RA patients were reduced with 25 and 50 μM PNLA by 60% (p &lt; 0.001) and in HC by 50% and 35% respectively (p &lt; 0.01). PGE2 levels were inhibited by the same percentage in both HC and RA monocytes (p &lt; 0.001) by 50 μM PNLA. Similarly, effects were observed with IL-6, TNF- α, and PGE2 levels in LPS-stimulated macrophages especially in RA patients 30% (p &lt; 0.05). Conclusion Our data suggest that PNLA significantly attenuated monocyte migration, significantly reduced macropinocytosis and Dil-oxLDL uptake in macrophages. Furthermore, PNLA inhibited production of IL-6, TNF-α and PGE2 levels in LPS-stimulated monocytes and macrophages from RA patients. These data inform on the potential anti-inflammatory and analgesic effects of PNLA. Disclosures R. Takala None. D. Ramji None. E. Choy None.

scholarly journals Anti-Rheumatic Effect of Antisense Oligonucleotide Cytos-11 Targeting TNF-α Expression

2021 ◽  
Vol 22 (3) ◽  
pp. 1022
Author(s):  
Tatyana P. Makalish ◽  
Ilya O. Golovkin ◽  
Volodymyr V. Oberemok ◽  
Kateryna V. Laikova ◽  
Zenure Z. Temirova ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Gene Expression ◽  
Rat Model ◽  
Peripheral Blood ◽  
Antisense Oligonucleotide ◽  
Joint Inflammation ◽  
Blood Concentrations ◽  
Tnf Α ◽  
Effective Drugs ◽  
First Time

The urgency of the search for inexpensive and effective drugs with localized action for the treatment of rheumatoid arthritis continues unabated. In this study, for the first time we investigated the Cytos-11 antisense oligonucleotide suppression of TNF-α gene expression in a rat model of rheumatoid arthritis induced by complete Freund’s adjuvant. Cytos-11 has been shown to effectively reduce peripheral blood concentrations of TNF-α, reduce joint inflammation, and reduce pannus development. The results achieved following treatment with the antisense oligonucleotide Cytos-11 were similar to those of adalimumab (Humira®); they also compared favorably with those results, which provides evidence of the promise of drugs based on antisense technologies in the treatment of this disease.

scholarly journals Anti-Inflammatory Effect and Cellular Uptake Mechanism of Carbon Nanodots in in Human Microvascular Endothelial Cells

Nanomaterials ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1247
Author(s):  
Sarah Belperain ◽  
Zi Yae Kang ◽  
Andrew Dunphy ◽  
Brandon Priebe ◽  
Norman H. L. Chiu ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Interleukin 1 ◽  
Antioxidant Properties ◽  
Synthesis Method ◽  
Microvascular Endothelial Cells ◽  
Carbon Nanodots ◽  
Uptake Mechanism ◽  
Tnf Α ◽  
Anti Inflammatory ◽  
Microvascular Endothelial

Cardiovascular disease (CVD) has become an increasingly important topic in the field of medical research due to the steadily increasing rates of mortality caused by this disease. With recent advancements in nanotechnology, a push for new, novel treatments for CVD utilizing these new materials has begun. Carbon Nanodots (CNDs), are a new form of nanoparticles that have been coveted due to the green synthesis method, biocompatibility, fluorescent capabilities and potential anti-antioxidant properties. With much research pouring into CNDs being used as bioimaging and drug delivery tools, few studies have been completed on their anti-inflammatory potential, especially in the cardiovascular system. CVD begins initially by endothelial cell inflammation. The cause of this inflammation can come from many sources; one being tumor necrosis factor (TNF-α), which can not only trigger inflammation but prolong its existence by causing a storm of pro-inflammatory cytokines. This study investigated the ability of CNDs to attenuate TNF-α induced inflammation in human microvascular endothelial cells (HMEC-1). Results show that CNDs at non-cytotoxic concentrations reduce the expression of pro-inflammatory genes, mainly Interleukin-8 (IL-8), and interleukin 1 beta (IL-1β). The uptake of CNDs by HMEC-1s was examined. Results from the studies involving channel blockers and endocytosis disruptors suggest that uptake takes place by endocytosis. These findings provide insights on the interaction CNDs and endothelial cells undergoing TNF-α induced cellular inflammation.

scholarly journals Camel Milk Attenuates Rheumatoid Arthritis Via Inhibition of Mitogen Activated Protein Kinase Pathway

2017 ◽  
Vol 43 (2) ◽  
pp. 540-552 ◽  
Author(s):  
Hany H. Arab ◽  
Samir A. Salama ◽  
Tamer M. Abdelghany ◽  
Hany A. Omar ◽  
El-Shaimaa A. Arafa ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Mapk Pathway ◽  
Lipid Peroxide ◽  
Mitogen Activated Protein Kinase ◽  
Camel Milk ◽  
Tnf Α ◽  
Mitogen Activated Protein ◽  
Anti Oxidant ◽  
Cox 2 ◽  
Anti Inflammatory

Background/Aims: Camel milk (CM) has shown beneficial anti-inflammatory actions in several experimental and clinical settings. So far, its effect on rheumatoid arthritis (RA) has not been previously explored. Thus, the current work aimed to evaluate the effects of CM in Adjuvant-induced arthritis and air pouch edema models in rats, which mimic human RA. Methods: CM was administered at 10 ml/kg orally for 3 weeks starting on the day of Freund’s adjuvant paw inoculation. The levels of TNF-α and IL-10 were measured by ELISA while the protein expression of NF-κBp65, COX-2 and iNOS was detected by immunohistochemistry. The expression of MAPK target proteins was assessed by Western blotting. Results: CM attenuated paw edema, arthritic index and gait score along with dorsal pouch inflammatory cell migration. CM lowered the TNF-α and augmented the anti-inflammatory IL-10 levels in sera and exudates of arthritic rats. It also attenuated the expression of activated NF-κBp65, COX-2 and iNOS in the lining of the dorsal pouch. Notably, CM inhibited the MAPK pathway signal transduction via lowering the phosphorylation of p38 MAPK, ERK1/2 and JNK1/2 in rat hind paws. Additionally, CM administration lowered the lipid peroxide and nitric oxide levels and boosted glutathione and total anti-oxidant capacity in sera and exudates of animals. Conclusion: The observed CM downregulation of the arthritic process may support the interest of CM consumption as an adjunct approach for the management of RA.

Oxymatrine inhibits lipopolysaccharide-induced inflammation by down-regulating Toll-like receptor 4/nuclear factor-kappa B in macrophages

2015 ◽  
Vol 93 (4) ◽  
pp. 253-260 ◽  
Author(s):  
Yu Zhang ◽  
Ruhong Yan ◽  
Yae Hu
Keyword(s):  
Nitric Oxide ◽  
Interleukin 1 ◽  
Chinese Herb ◽  
Mrna Levels ◽  
Nuclear Factor ◽  
Toll Like Receptor ◽  
Toll Like Receptor 4 ◽  
Necrosis Factor Alpha ◽  
Tnf Α ◽  
Anti Inflammatory

Oxymatrine (OMT) is the quinolizidine alkaloid extracted from the Chinese herb Sophora flavescens Ait. that has many pharmacological effects and is used for the treatment of some inflammatory diseases. In this study, RAW264.7 cells and THP-1 differentiated macrophages were pretreated with various concentrations of OMT at 2 h prior to treatment with lipopolysaccharide (LPS) (1.0 μg/mL) for different durations. We detected the anti-inflammatory effect of OMT in LPS-stimulated macrophages and investigated the molecular mechanism. We showed that OMT pretreatment significantly inhibited the LPS-induced secretion of nitric oxide (NO), interleukin-1 beta (IL-1β), and tumor necrosis factor-alpha (TNF-α) in supernatant, attenuated the mRNA levels of inducible nitric oxide synthase (iNOS), IL-1β, TNF-α, and Toll-like receptor 4 (TLR4), increased TLR4 and phosphorylation of inhibitor of kappa B-alpha (p-IBα) in cytosol, and decreased the nuclear level of nuclear factor-κB (NF-κB) p65 in macrophages. In conclusion, OMT exerts anti-inflammatory properties in LPS-stimulated macrophages by down-regulating the TLR4/NF-κB pathway.

scholarly journals Inflammatory Cytokine Neutralizing Antibody Treatment Could Recover Increased Follicular Helper and Follicular Regulatory T Cells in Murine Models of Arthritis

2021 ◽  
Author(s):  
Xingyue Zeng ◽  
Meng Li ◽  
Mohan Zheng ◽  
Tianci Liu ◽  
Rui Kang ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Peripheral Blood ◽  
Neutralizing Antibody ◽  
Mouse Spleen ◽  
Inflammatory Factor ◽  
Antibody Treatment ◽  
Follicular Helper ◽  
Anti Inflammatory ◽  
And Function ◽  
The Impact

Abstract Objective. We aimed to illustrate the changes of follicular helper (TFH) and follicular regulatory (TFR) cells in rheumatoid arthritis using the collagen-induced arthritis (CIA) model and clarity the impact of anti-inflammatory treatment on TFH and TFR cells.Methods. We established 10-week-old CIA model and used flow cytometry to analyze the changes of TFH and TFR cells in peripheral blood and spleen at different time points (5w, 7w, 10w,13w). The expression of TIGIT, CD226, ICOS and PD-1 characterizing the functions of TFH and TFR were also analyzed. The function of spleen TFH and TFR cells from CIA was further analyzed. The effects of anti-inflammatory antibody treatments on the subpopulation and function changes of TFH and TFR were also analyzed in CIA mice.Results. The levels of TFH and TFR were significantly increased in spleen and peripheral blood of CIA mice. After treatment, TFH and TFR cells decreased significantly. TIGIT+ and TIGIT+CD226-TFH cells in CIA mouse spleen were elevated and PD-1 and ICOS expression on TFH and TFR cells in the spleen were also significantly increased. The ability of TFH to secrete IL-21 and help B cells, and TFR to secrete IL-10 and inhibit TFH were both enhanced in CIA mouse spleen. After antibody treatment, cell subsets and functions were significantly recovered.Conclusion. In the pathogenesis of rheumatoid arthritis, TFH and TFR cells in the germinal center increases and their functions are enhanced. After the treatment by inflammatory factor antibodies, TFH and TFR subsets and their functions can be significantly recovered.

scholarly journals Sec-O-glucosylhamaudol suppressed inflammatory reaction induced by LPS in RAW264.7 cells through inhibition of NF-κB and MAPKs signaling

2020 ◽  
Vol 40 (2) ◽  
Author(s):  
Guiming Liu ◽  
Jing Xie ◽  
Yurui Shi ◽  
Rongda Chen ◽  
Li Li ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Bioactive Compound ◽  
Therapeutic Effects ◽  
Raw 264.7 ◽  
Pcr Assay ◽  
Raw 264.7 Cell ◽  
Tnf Α ◽  
Anti Inflammatory ◽  
Saposhnikovia Divaricata ◽  
Inflammation Cytokines

Abstract As a major bioactive compound from the Saposhnikovia divaricata (Turcz.) Schischk, sec-O-glucosylhamaudol (SOG), has been reported to have anti-nociceptive activity and high 5-lipoxygenase (5-LOX) activity. Nevertheless, the mechanism of the potential anti-inflammatory effects of SOG is unclear. The anti-inflammatory impacts of SOG in RAW 264.7 cell lines stimulated by LPS were explored in the present study. It was found that SOG dose-dependently reduced the emergence of inflammation cytokines, such as IL-6 and TNF-α in Raw264.7 murine macrophages stimulated by LPS. Real-time PCR assay demonstrated the SOG dose-dependently inhibited transcription of these cytokines as well. In addition, it was also found that NF-κB activation and MAPKs phosphorylation including p38, JNK and ERK1/2 induced by LPS were suppressed by SOG. Due to its anti-inflammatory activity, our results suggest that SOG might have therapeutic effects on inflammatory disease, such as acute lung injury or rheumatoid arthritis.

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