Induced Arthritis Model Research Articles

Discovery and pre-clinical characterization of a selective PI3Kδ inhibitor, LL-00071210 in rheumatoid arthritis

PI3Kδ plays a critical role in adaptive immune cell activation and function. Suppression of PI3Kδ has been shown to counter excessive triggering of immune responses which has led to delineating the role of this isoform in the pathophysiology of autoimmune disorders. In the current study, we have described preclinical characterization of PI3Kδ specific inhibitor LL-00071210 in various rheumatoid arthritis models. LL-00071210 displayed excellent in vitro potency in biochemical and cellular assay against PI3Kδ with IC50 values of 24.6 nM and 9.4 nM, respectively. LL-00071210 showed higher selectivity over PI3Kγ and PI3Kβ as compared to available PI3K inhibitors. LL-00071210 had good stability in liver microsomes and plasma across species and showed low clearance, low-to-moderate Vss, with bioavailability of >50% in preclinical species. LL-00071210 demonstrated excellent in vivo efficacy in adjuvant-induced and collagen-induced arthritis models. Co-administration of LL-00071210 and methotrexate at subtherapeutic dose regimen in collagen induced arthritis model led to additive effects, indicating the combination potential of LL-00071210 along with available disease modifying anti-rheumatic drugs (DMARD). In conclusion, we have described a specific PI3Kδ inhibitor with ∼100-fold selectivity over other PI3K isoforms. LL-00071210 has good drug-like properties and thus warrants testing in the clinic for the treatment of autoimmune diseases.

Intra-articular injection of monoiodoacetate induces diverse hip osteoarthritis in rats, depending on its dose

BackgroundMonoiodoacetate (MIA)-induced arthritis models are used widely in osteoarthritis (OA) research to develop effective conservative treatments for hip OA, as an alternative to joint replacement surgery. In joint OA models, such as the MIA-induced knee OA model, various doses of MIA are utilized, depending on the purpose of the research. So far, only 2 mg of MIA has been used for MIA-induced hip OA research. We hypothesized that the amount of MIA should be adjusted according to the osteoarthritis model under investigation. We performed radiographic and histological evaluations in rats for hip OA models induced by different doses of MIA.MethodsOne hundred and eighty right hips of six-week-old, male Sprague–Dawley rats (n = 30 rats per group) were treated with either a single intra-articular injection of various doses of MIA (0.25, 0.5, 1.0, 2.0, and 4.0 mg) dissolved in 25 μl of sterile saline (MIA group), or with 25 μl of sterile saline alone (Sham group). Radiographic and histological evaluations of the hip joint were performed at one, two, four, eight, and 12 weeks after administration (n = 6 rats per group per time point).ResultsOA changes progressed from 1 week after administration in the 1.0-mg, 2.0-mg, and 4.0-mg MIA groups. The degree of OA changes increased as the dose of MIA increased. The 0.25-mg and 0.5-mg MIA groups presented fewer OA changes than the 2.0-mg and 4.0-mg MIA groups during the entire study period (up to 12 weeks). The administration of 0.25 mg and 0.5 mg of MIA-induced both radiographic and histological OA changes in a time-dependent manner, whereas more than 2 mg of MIA provoked end-stage OA at 8 weeks after injection. Absolute, dose-dependent histopathological OA changes were observed 4 weeks after MIA administration.ConclusionsIntra-articular MIA injection to the hip joints of rats induced diverse OA changes dose-dependently. Research for developing novel conservative treatments for hip OA and intractable pain should consider the pathological condition when determining the dose of MIA to be employed.

Pterostilbene exert an anti-arthritic effect by attenuating inflammation, oxidative stress, and alteration of gut microbiota.

Pterostilbene is a revesterol analog with a long bioavailability and having potent anti-inflammatory activity in animal studies. In this study, we tried to scrutinize the anti-arthritic effect of pterostilbene against complete Freund's adjuvant (CFA)-induced arthritis model in rats. CFA was used for induction of the RA, and rats were divided into groups depending on different doses of pterostilbene given. Hepatic, antioxidant, rheumatoid factor (RF), myeloperoxidase (MPO), inflammatory cytokines, anti-collagen(C)II-Ig, inflammatory mediators, and gut microbiota were estimated. Pterostilbene significantly (p<.001) decreased the paw swelling, arthritic score, and increased the body weight. Besides altered the antioxidant, inflammatory mediators, anti-collagen (C)II Ig, and inflammatory cytokines. Furthermore, pterostilbene treatment helps to restore the ecosystem of gut microbiota in rats by reducing the relative abundance of Helicobacter, Desulfovibrio, Lachnospiraceae, and Mucispirillium. Based on the findings, we can say that pterostilbene has an anti-arthritic effect via suppressing inflammatory responses and altering intestinal bacteria. PRACTICAL APPLICATIONS: Arthritis is the painful disease and affected most of the people worldwide. In this experimental study, we estimated the anti-arthritic effect of pterostilbene against CFA-induced arthritis in rats. Pterostilbene noticeably suppressed the paw thickness, arthritic score, and organ index. Pterostilbene substantially altered the oxidative stress and inflammatory reaction. Pterostilbene considerably modulated the gut microbiota, suggesting the anti-arthritic effect.

Anti-arthritic activity of the flavonoids fraction of ivy leaves (Hedera helix L.) standardized extract in adjuvant induced arthritis model in rats in relation to its metabolite profile using LC/MS.

Ivy leaves (Hedera helix) is a traditional plant used for common cold, cough, and bronchial disorders and can be used for rheumatoid arthritis (RA) as an attempt in alternative medicine. RA is a chronic autoimmune disease characterized by its increasing frequency and adverse consequences. There is an urgent need for a long-term therapy that has favorable biological effects and is less expensive than the already authorized synthetic medicines. This study aimed to determine the anti-arthritic potentials of Hedera helix with determination of the bioactive fraction and discovery of its second-generation metabolites by means of LC/MS. The total ivy ethanolic extract (TIE-E), saponins fraction (Sap-F) and flavonoids fraction (Flav-F) were investigated for their in-vitro anti-arthritic effects and in-vivo by Adjuvant-induced arthritis (AIA) using Complete Freund's Adjuvant (0.1mL, CFA) intradermal relative to the usual dose of ibuprofen (5mg/kg). We examined the physical alterations, rheumatoid biomarkers, cytokines that cause and inhibit inflammation, markers of oxidative stress, hyaluronidase and β-glucuronidase enzyme activity. Each paw's histopathology was also evaluated. The chemical profiles of TIE-E were studied using LC/MS in both positive and negative ionization modes. TIE-E (200mg/kg) and Flav-F (100mg/kg) significantly (P< 0.05) lowered the edema of the paws, serum immunological indicators, inflammatory cytokines, degenerative enzymes, and indicators of reactive oxygen species with increasing in the anti-inflammatory cytokines. Our findings suggest that extracts of ivy leaves might be used effectively to treat rheumatoid arthritis, where its flavonoid content is responsible for that, and it is able to repress biochemical, oxidative, and pathological changes associated with (AIA) Adjuvant-induced arthritis.

Anti-Inflammatory Effects of BoNT/A Against Complete Freund's Adjuvant-Induced Arthritis Pain in Rats: Transcriptome Analysis.

Arthritis is the most common cause to lead to chronic pain. Botulinum toxin type A (BoNT/A) has been widely used to treat chronic pain. In our previous study, we confirmed the anti-inflammatory and antinociceptive effects of BoNT/A in the Complete Freund’s Adjuvant (CFA)-induced arthritis model, but the underlying anti-inflammatory mechanism was not fully elucidated. The purpose of this study was to investigate the anti-inflammatory effects and mechanisms of BoNT/A on arthritis using transcriptomic analysis. The BoNT/A was injected into the rat ankle joint on day 21 after CFA injection. The von Frey and hot plate tests were applied to assess the pain-related behaviors at different time points. Five days after BoNT/A treatment, gene expression profiling in dorsal root ganglion (DRG) was performed using RNA sequencing (RNA-seq). The differentially expressed genes (DEGs) were analyzed by various tools. The mechanical allodynia and thermal hyperalgesia were significantly reversed after BoNT/A injection. RNA-seq revealed 97 DEGs between the CFA group and Sham group; these DEGs were enriched inflammatory response, IL-17 signaling pathway, etc. There are 71 DEGs between the CFA+BoNT/A group and the CFA group; these DEGs related to response to peptide, PI3K-Akt signaling pathway, ECM–receptor interactions, etc. Three key genes were significantly decreased after CFA-induced arthritis pain, while BoNT/A increased the expression of these genes. The identification of S100A9, S100A8, and MMP8 genes can provide new therapeutic targets for arthritis pain and affect the signaling pathway to play an anti-inflammatory role after the treatment of BoNT/A.

Effect of Orally-administration Niclosamide on serum level of VCAM-1 and E-selectin in Collagen induced arthritis: Experimental study

Soluble adhesion molecules (mainly VCAM-1 and E-selectin) have a vital role in the pathogenesis of the rheumatoid arthritis (RA) and consider as angiogenic mediators for this disease. The main goal for this research is to evaluate the efficacy of orally administer niclosamide (NCS) in prevention the angiogenic mediators (VCAM-1 and E-selectin) using collagen induced arthritis model in rats (CIA). Fifty male Spraque-Dawley rats underwent collagen induced arthritis (CIA) model. When arthritis was fully developed, the rats were either treated orally with low-dose (50mg/kg) NCS or high-dose (100mg/kg) NCS or treated intrapertonially (IP) with 30mg/kg NCS or leave without treatment for 4 weeks. Body weight measurement and arthritis index were monitor before and after treatment in all groups. At the end of the treatment period serum level of vascular cell adhesion molecule-1 (VCAM-1), E-selectin and TNFα were measured together with collection of articular synovial tissue to evaluate the pathological changes. The experiment showed that NCS significantly reduce the arthritis index, foot pad thickness and ankle swelling (p value &lt; 0.05) when given orally in a high dose and IP to the experimental animal. Comparing to the CIA model group, significant reduction in the serum level of VCAM1 and E-selectin has been observed in those rats treated with high dose of oral NCS or IP injection of NCS (p value &lt; 0.05). Niclosamide can effectively decrease, in dose dependent manner, the clinical scores, joint swelling, VCAM1, E-selectin and pathological changes in arthritic rats induced by collagen type II.

Pharmacological and Biochemical evaluation of anti-arthritic activity of Justicia gendarussa extract in FCA induced arthritis in Wistar rats

The present study is carried out to evaluate anti-inflammatory and anti-arthritic potential of Justicia gendarussa. Leaves and stem extracted with ethanol and chloroform. These extract is tested against in-vitro (HRBC membrane stabilization method and Inhibition of protein denaturation method) and in-vivo (Carrageenan induced paw edema model) anti-inflammatory models. To assess the anti-arthritic activity, FCA induced arthritis model is used. Assessment of arthritis is done by paw volume, joint diameter, body weight, biochemical parameters, hematological parameters (Hb, RBC, WBC, ESR). The effect of in-vitro anti-inflammatory model depends on concentration. Both test extract and standard Diclofenac sodium has been shown concentration dependent effect. The maximum anti-inflammatory effect of the test extract achieved at 2000μg/ml. The test (JGLE, JGLC, JGSE) has been shown inhibition of paw edema induced by carrageenan at 50mg/kg body weight. The extract JGLE, JGLC, JGSE at 50mg/kg body weight and 100mg/kg body weight orally showed the significant (P&lt; 0.05) and dose dependent inhibitory effect against FCA induced arthritis model. Diclofenac sodium 20mg/kg body weight orally is used as a standard. JGLE exhibit more significant and most promising anti-arthritic and anti-inflammatory effect than other extracts these effects support the traditional role of J. gendarussa in arthritis and other inflammatory condition.

A 8-mer Peptide of PGLYRP1/Tag7 Innate Immunity Protein Binds to TNFR1 Receptor and Inhibits TNFα-Induced Cytotoxic Effect and Inflammation.

Search for novel regulatory protein fragments with potential functional roles is required both for understanding the immune response mechanisms and the development of targeted immunotherapy. Earlier we demonstrated that the PGLYRP1/Tag7 innate immunity protein can be regarded as an inhibitor of TNFα cytotoxic activity via the interaction with its TNF receptor 1 (TNFR1). A C-terminal peptide fragment 17.1 of the molecule is responsible for this function. In this study we have identified a minimal 8-mer region of this peptide (hereinafter – 17.1A) capable to bind to TNFR1. As a result of such interaction, the cytotoxic signals induced by this receptor are blocked. Also, this peptide demonstrates an anti-inflammatory activity in vivo in the complete Freund’s adjuvant (CFA)-induced arthritis model in laboratory mice. Peptide 17.1A is capable to reduce periarticular inflammation, inhibit the development of synovitis and exhibit a protective effect on cartilage and bone tissues. This peptide can turn out to be a promising medicinal agent for autoimmune arthritis and other diseases.

Investigation of anti-inflammatory and anti-arthritic potentials of Terminalia catappa bark using in vitro assays and carrageenan-induced inflammation, complete Freund's adjuvant induced arthritis model in rats

Terminalia catappa Linn (T. catappa) belongs to the family Combretaceae, locally known as Bengal Almond. The plant is known to have many phytochemicals such as carbohydrates, phytosterols, tannins, polyphenols, flavonoids, and saponins. It is widely used in the treatment of various diseases such as dermatitis, leprosy, and scabies and internally for colic, headache and hepatitis and also, the plant possesses anticancer, antioxidant, antidiabetic, hepatoprotective and extracts have been shown to modify reproductive behavior in rats. The main objective of this study is to assess the pharmacological potentials of T. catappa bark in the therapeutic areas of inflammation and arthritis. T. catappa bark alcoholic extract (TCE) and water extract (TCW) were prepared and evaluated for its possible anti-inflammatory activity by in vitro protein denaturation assays and membrane stabilization assay. Further, the anti-inflammatory and anti-arthritic activity of bark extracts were evaluated by in vivo models such as carrageenan-induced paw edema, air-pouch model and Complete Freund's adjuvant (CFA) induced arthritis model in rats. Total phenolic content (TPC) was found to be 287 and 175 mg/g for TCE and TCW. Total flavonoid content (TFC) was found to be 10.2 and 61.7 mg/Kg for TCE and TCW. The concentration required for the 50 % inhibition of the protein denaturation (IC 50) was found to be lower for TCE (36.2 µg/mL) in egg albumin denaturation assay, whereas in the membrane stabilization assay method TCW exhibited a lower IC50 value in comparison with diclofenac. Both the extracts showed activity in the in vitro models of inflammation and were also found to be effective in reducing the paw edema in the carrageenan-induced inflammation model and inhibition of cellular infiltration and Myeloperoxidase activity in air pouch fluid, suggesting its anti-inflammatory activity. In CFA-induced arthritis model, the extracts exhibited anti-arthritic potential by improving the altered blood parameters, arthritic scores, radio-graphical, and histopathological changes. The results of the study suggested that the bark of T. catappa possessed anti-inflammatory and anti-arthritic activities.

The preventive potential of Citrullus vulgaris (Thunb.) seed extract on cafeteria diet, atherogenic diet-induced obesity, and formaldehyde/ Freund's adjuvant arthritis models in rats

This present study was aimed to explore the anti obesity and anti arthritic activities of seed extracts of Citrullus vulgaris (Thunb) in cafeteria diet (CD), atherogenic diet (AD) induced obesity and Formaldehyde and Freund’s adjuvant (FA) induced arthritis in experimental animals i.e. rats. The ethanolic and aqueous extracts were prepared by using soxhlet apparatus and maceration process respectively. Anti obesity activity of seed extracts of this plant was evaluated in CD and AD induced obesity and anti arthritic activity was evaluated in formaldehyde and FA induced arthritis in rats. Phytochemical studies with both ethanolic extract (ELSCV) and aqueous extract (AQSCV) revealed the presence of carbohydrates, proteins, flavonoids, saponins, fixed oils, glycosides and steroids. No mortality or behavioral abnormality recorded in mice at the highest dose level of 2000 mg/kg tested for LD50 studies. Both the extracts with medium and high doses exhibited a significant anti obesity activity by reducing the body weight, food intake, organ and fat pads weight and serum glucose, cholesterol, triglyceride, low density lipoproteins and very low density lipoprotein cholesterol levels with an increased high density levels in CD and AD induced obesity models in rats. Both extracts with medium and high doses exhibited a significant anti arthritic activity and reduced serum biochemical parameters in FA induced arthritis model in rats. The anti obesity and anti arthritic activities with extracts conformed the above mentioned activities because presence of flavonoids, saponins, fixed oils, steroids, alkaloids and glycosides.

Activation of the Nrf2/HO-1 signaling pathway by dimethyl fumarate ameliorates complete Freund's adjuvant-induced arthritis in rats

The nuclear factor erythroid 2-related factor (Nrf2) signaling pathway has recently emerged as a novel therapeutic target in treating various diseases. Therefore, the present study aimed to assess the protective role of the Nrf2 activator, dimethyl fumarate (DMF) in the complete Freund's adjuvant (CFA)- induced arthritis model. DMF (25, 50, and 100 mg/kg) and dexamethasone (2 mg/kg) were orally administered for 14 days. Pain-related tests, paw volume, and arthritic scores were measured weekly. Serum TNF-α, IL-1β, cyclic citrullinated peptide (CCP), C-reactive protein (CRP), and rheumatoid factor (RF) levels were estimated. Nitrite, malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione (GSH), catalase (CAT), and myeloperoxidase (MPO) levels were also evaluated. NF-κB, Nrf2, HO-1, and COX-2 levels were estimated in the joint tissue. DMF treatment exerted anti-arthritic activity by enhancing the nociceptive threshold, improving arthritis scores, and reducing paw edema. Also, DMF suppressed changes in oxidative stress markers and inflammatory mediators and enhanced Nrf2 and HO-1 levels in CFA-injected rats. These findings indicate that the anti-arthritic activity of DMF may be mediated by the activation of the Nrf2/HO-1 pathway, which reduced oxidative damage and inflammation.

Pharmacological screening of ethanolic extract of Pithecellobium dulce for antiarthritic activity in Rats

Arthritis is an inflammatory joint disorder in which cartilage of the joint is gradually lost and categorized by swelling of joints, pain, and loss of function. The present study was conducted to explore the antiarthritic activity of ethanolic extract of Pithecellobium dulce against formaldehyde induced at sub plantar region of the left hind paw during days 1 and 3 of the study period. The changes observed in paw diameter during the study period, various biochemical, and haematological parameters were monitored. The Radiographic analysis and histopathology significantly improved after treatment with test extract P. dulce (250 mg/kg, b.w., p.o. ) as compared to the standard treatment with indomethacin (10 mg/kg, b.w., p.o. ).The results of the current investigation concluded that ethanolic extract of P. dulce possesses significant anti-arthritic activity against formaldehyde induced arthritis model, justifying its therapeutic role in arthritic conditions. The observed antiarthritic activity may be due to the presence of phytoconstituents such as alkaloids and flavonoids. P. dulce significantly suppressed the paw oedema in formaldehyde models ( P &lt;0.001). The Histopathological and radiographic studies of joints also showed a protective effect of P. dulce .

Evaluation of Prophylactic Diacerein Treatment on Anti-Arthritic Activity in Freund’s Complete Adjuvant-Induced Arthritis in Rat Model

Introduction: Osteoarthritis (OA) is commonly prevalent disorder seen in Indian population. Nonsteroidal Anti-Inflammatory Drugs (NSAIDS) are used as primary drugs for its symptomatic treatment. However, its use is limited by its long-term adverse effects. Many clinical studies have shown that diacerein reduces clinical symptoms of OA. Aim: To evaluate whether diacerein has beneficial anti-arthritic property when used prophylactically in Freund’s Complete Adjuvant (FCA) induced arthritis in rats. Materials and Methods: An experimental study was conducted over a period of about two months in the Department of Pharmacology, SCBMC, Cuttack. Thirty albino rats of Wistar strain was divided into 5 groups with 6 animals in each group. The basal body weight and the hind paw volumes of both right and left paw of all the animals was noted in day 0 and then on 4th, 8th, 14th and 21st day. Arthritis was induced in all animals by injecting FCA on day 0 into the plantar surface of right hind paw. Normal saline, diclofenac 5 mg/kg, three doses of diacerein (50 mg/kg, 100 mg/kg, 200 mg/kg), was administered orally once daily to groups I, II, III, IV and V respectively. Paw volumes and body weight was measured for arthritic parameters. On day 22 radiological and histopathologic evaluation was done. Results: Maximum inhibition of Freund’s adjuvant induced arthritic paw volume was on the 21st day with 100 mg/kg of diacerein and with diclofenac 5 mg/kg was from 8th to 14th day. There was a decrease in body weight due to freund’s adjuvant in normal saline group from 4th to 21st day, still greater decline with diclofenac 5 mg/kg group, but in diacerein treated group there was an increase in body weight from 4th to 21st day in all the three doses. Radiologically and histopathology maximum benefit was noted with 100 mg/kg of diacerein. Conclusion: Three weeks treatment of oral diacerein can significantly inhibit arthritic swelling of the injected paw at all the 3 doses in adjuvant induced arthritis model in rats and can be a promising disease modifying drug for OA.

Metabolomic and transcriptomic analyses of the anti-rheumatoid arthritis potential of xylopic acid in a bioinspired lipoprotein nanoformulation

Xylopic acid (XA), a diterpene kaurene and the major active ingredient of the African spice Xylopia aethiopica (Annonaceae), is reported to possess anti-inflammatory and analgesic properties. Here, we investigated the therapeutic potential of XA for rheumatoid arthritis (RA), a debilitating autoimmune inflammatory disease characterized by joint damage, in the complete Freund's adjuvant (CFA)-induced arthritis model in rats. We synthesized bioinspired reconstituted high-density lipoprotein (rHDL) nanoparticles loaded with purified XA crystals (rHDL/XA) that passively accumulate in inflamed joints of CFA-induced arthritic rats. Treatment with rHDL/XA minimized mononuclear cell infiltration of CFA-induced arthritic sites and ameliorated disease burden. Metabolomic and transcriptomic analyses revealed that the major molecular pathways perturbed following CFA-induced arthritis correlated with amino acid and lipid metabolism, which were restored to normal states by rHDL/XA treatment. This work demonstrates the anti-RA potential of XA in a nanoformulation and uncovers its underlying therapeutic mechanisms at the transcript and metabolite levels.

Tunisian Olea europaea L. leaf extract suppresses Freund's complete adjuvant-induced rheumatoid arthritis and lipopolysaccharide-induced inflammatory responses

Ethnopharmacological relevanceOlea europaea L. (olive) is traditionally used as a folk remedy and functional food in Europe and Mediterranean countries to treat inflammatory diseases. O. europaea contains phenolic compounds and have been reported to prevent cartilage degradation. However, the function and mechanism of O. europaea in rheumatoid arthritis are not known.Aim of the study: In this study, we aimed to examine anti-inflammatory and anti-arthritic effects of Tunisian O. europaea L. leaf ethanol extract (Oe-EE). Materials and methodsTo do this, we employed an in vitro macrophage-like cell line and an in vivo Freund's complete adjuvant (AIA)-induced arthritis model. Levels of inflammatory genes and mediators were determined from in vivo samples. ResultsThe Oe-EE clearly reduced the production of the lipopolysaccharide-mediated inflammatory mediators, nitric oxide (NO) and prostaglandin E2 (PGE2), in RAW264.7 cells. The results of HPLC showed that Oe-EE contained many active compounds such as oleuropein and flavonoids. In AIA-treated rats, swelling of paws, pain, and cartilage degeneration were alleviated by oral Oe-EE administration. Correlating with in vitro data, PGE2 production was significantly reduced in paw samples. Furthermore, the molecular mechanism of Oe-EE was dissected, and Oe-EE regulated the gene expression of interleukin (IL)-6, inducible NO synthase (iNOS), and MMPs and inflammatory signaling activation. ConclusionConsequently, Oe-EE possesses anti-inflammatory and anti-rheumatic effects and is a potential effective treatment for rheumatoid arthritis.

Sesquiterpene lactones; Damsin and neoambrosin suppress cytokine-mediated inflammation in complete Freund's adjuvant rat model via shutting Akt/ERK1/2/STAT3 signaling

Ethnopharmacologial relevanceAlthough Damsissa (Ambrosia maritima) is traditionally used as anti-inflammatory and diuretic, the biological activity and mechanism of action of its major constituents are to be elucidated. Aimto decipher the anti-arthritic potential of damsin (DMS) and neoambrosin (NMS) and to unfold their molecular signaling in complete Freund's adjuvant (CFA)-induced arthritis model. Materials and methodsthe right hind paw was inoculated with CFA (0.1 ml) at day 0 and 7 while treatments were started from the 14th day and continued for 2 weeks. Rats were randomly assigned into 4 groups; normal group (NRML), CFA-induced arthritis group, CFA-induced arthritis treated with DMS and NMS (10 mg/kg/day) as 3rd and 4th group; respectively. ResultsThroughout experimental period, treatments ameliorated the increase of paw volume, knee joint diameter and nociception tests as reflected in open field arena. Also, DSM and NMS suppressed phosphorylation of Akt, STAT-3, ERK1/2 which was further mirrored by inactivation of GSK3β and downregulation of MCP-1 together with CCN1 and NF-kβ in hind paw tissue. Concomitantly, inflammation markers; TNF-α, IL-6, -12 were lowered as confirmed microscopically during examination of hind paw tissue. ConclusionDSM and NMS-induced suppression of NF-kβ subdues clinical features of RA most probably through repression of Akt/ERK1/2/STAT3 pathway. Therefore, DMS and NMS can serve as safe and effective treatment for rheumatoid arthritis, one of the most disabling chronic, inflammatory and painful autoimmune disease.

Interferon gamma/interleukin-4 modulation, anti-inflammatory and antioxidant effects of hesperidin in complete Freund’s adjuvant (CFA)-induced arthritis model of rats

Background This study sought to assess the effect of hesperidin on serum inflammatory cytokines and oxidative damage in liver of complete Freund’s adjuvant (CFA)-induced arthritic rats. Method Fifty-six adult female Wistar rats (220–250 g) were acclimatized for two weeks. Intraplantar injection of CFA was done for the induction of arthritis and confirmed on the 14th day prior to oral administration of 40 and 80 mg/kg of hesperidin or dexamethasone for 45 days. Result The result showed that treatment with both doses of hesperidin and dexamethasone in the joint of arthritic rats significantly (p < .05) diminished paw swelling/edema and arthritis score as well as enhanced latency in thermal hyperalgesia test. In addition, hesperidin treatment in arthritis rats showed significant (p < .01) improvement in red blood cells and platelets counts as well as hemoglobin and hematocrit compared to the arthritis control rat group. Furthermore, hesperidin treatment significantly (p < .05) reduced serum interferon gamma (IFN-γ) and interleukin-4 (IL-4) levels in arthritic rat. In addition, treatment with hesperidin significantly (p < .05) decreased the liver of thiobarbituric acid reactive species and reactive oxygen species levels but raised the levels of total and non-protein thiols of rat induced with CFA. The reduced activities of liver δ-aminolevulinate dehydratase, catalase, glutathione-S transferase in arthritic rats were significantly (p < .05) increased with hesperidin treatment in arthritic rats. This study suggests that hesperidin demonstrated an anti-arthritic effect via modulation of serum IFN-γ and IL-4 levels as well as protection against oxidative damage. Conclusion Hence, hesperidin could be a potential immune-modulatory, anti-inflammatory and anti-oxidant agent.

Anti-inflammatory effect of stevioside abates Freund's complete adjuvant (FCA)-induced adjuvant arthritis in rats.

Adjuvant arthritis is a chronic, autoimmune and inflammatory disorder of the joints. The occurrence of disorder causes a severe damage to the connective tissue eventually leading to progressive physical disability and eventual death. The recent years of evidence suggests the anti-inflammatory properties of stevioside, a diterpene glycoside. However, the effect of stevioside against adjuvant arthritis, a chronic inflammatory disorder is not known. Hence, the present study was designed to study the effect of stevioside against Freund's complete adjuvant induced arthritis model in rats. The acute anti-inflammatory effect of stevioside also studied by employing carrageenan-induced paw oedema model in rats. The biochemical markers, haematological parameters, lipid peroxidation, myeloperoxidase activity, lipoxygenase activity, the levels of PGE2 and pro-inflammatory (TNF-α, IL-6 & IL-1β) and anti-inflammatory cytokine (IL-10) were analysed. The protein expression of NF-κB (p65) COX-2 and iNOS in paw tissues were estimated by western blotting. Stevioside treatment significantly ameliorates the adjuvant induced arthritic scoring, histological alterations, paw volume, elevation of biochemical (AST, ALT, ALP and glucose levels) and haematological (haemoglobin, differential and platelet count) parameters and restored the endogenous anti-oxidant (SOD, CAT, GSH and GST) activities. Treatment with stevioside also significantly prevented the adjuvant induced elevation of pro-inflammatory cytokines (TNF-α, IL-6 and IL-1β), pro-inflammatory protein expressions (iNOS, COX-2, NF-κB (p65) and pIκB/IκB ratio), prevented the increase in myeloperoxidase activity and significantly restored the anti-inflammatory (IL-10) cytokine level in paw tissues. Collectively, our findings suggest that stevioside may serve as anti-inflammatory agent and could serve as a potential adjunct therapeutic option in treating adjuvant arthritis.

To study the effect and mechanism of dimethyl fumarate [DMF] in adjuvant induced arthritis model in rats

Currently available disease modifying anti-rheumatoid drugs have limitations like dose-dependent toxicity and tolerance.Dimethyl fumarate has demonstrated anti-inflammatory and immunomodulatory properties in various animal models. Thus, thepresent study aimed to evaluate the effects and mechanism of DMF in a murine model of adjuvant-induced arthritis.A total of 84 rats were divided into early treatment groups (n=48) and late treatment groups (n=36). There were 8 subgroupsand 6 subgroups (n=6 in each group) in the early and late treatment groups, respectively. Experimental rheumatoid arthritis(RA) was induced in Wistar rats by injecting complete Freund's adjuvant (CFA) intradermally at the base of the tail. Antirheumatic effects were evaluated by arthritis and histopathological scoring of ankle joints. To evaluate anti-oxidant properties,GSH, catalase, SOD, and lipid peroxidation were measured. ESR, WBC count, TNF-α and IL-6 levels were measured to evaluatethe immunomodulatory properties of DMF. DMF demonstrated anti-inflammatory effects by decreasing arthritis andhistopathological scores compared to the CFA control group, though the difference was not statistically significant. DMFexhibited immunomodulatory properties as decreases in TLC count, serum TNF-α, and plasma IL-6 levels were observed. In allthe above-mentioned parameters, the best response was achieved with the early combination therapy of DMF 30 mg/kg andmethotrexate [Mtx] 0.1 mg/kg. In the present study, DMF demonstrated antirheumatoid effects in a rat model of CFA-inducedarthritis. The best antirheumatoid effect was achieved with the early combination of DMF and Mtx.

THU0081 MIR-17-5P REDUCES INFLAMMATION AND BONE EROSIONS IN COLLAGEN INDUCED ARTHRITIS MICE AND DIRECTLY TARGETS THE JAK-STAT PATHWAY IN RHEUMATOID ARTHRITIS FIBROBLAST-LIKE SYNOVIOCYTES.

Background:micro-RNAs (miR) are strong regulators of gene expression. Their involvement in RA key cytokines pathway regulation entitles them as important players in RA pathophysiology. The miR 17-92 cluster has been widely studied in cancer as they regulate cell apoptosis.Objectives:The aims of this study were to screen miR 17-92 cluster’s expression in different RA phenotypes (erosive and non erosive), further elucidate the mechanisms and direct targets involved in miR-17-5p anti-inflammatory role and to investigate miR-17-5p therapeutic effect in arthritis.Methods:A miR array was performed in synovial tissue from naïve erosive and non-erosive RA patients. Intra-articular delivery of miR-17 lipoplex was performed in collagen induced arthritis model in mice. Clinical, histological and structural effects were studied over the course of arthritis. In depth studies of miR-17 mechanisms of action were performed in primary RA-FLS isolated from RA synovial tissue.Results:Among others, miR-17-5p expression was reduced in erosive RA. miR-17 transfection in arthritic paws significantly reduced clinical inflammation. Moreover, synovial B cells, T cells, macrophages and polynuclear neutrophils infiltrates were significantly reduced. Structural damage was also decreased as shown by a reduction in the number of osteoclasts and erosion score by CT analysis. Pro-inflammatory cytokines of the IL-6 family, STAT3 target genes and IL-1β expression were also significantly reduced, but not TNF-alpha. miR17 directly targeted the 3’-untranslated region of STAT3 and JAK1. STAT3 and JAK1 mRNA and protein expression were reduced in RA-FLS following miR-17 transfection. STAT3 and JAK1 mRNA and activation of STAT3 as assessed by immunohistochemistry were also reduced in injected paws.Conclusion:We demonstrate an anti-inflammatory and anti-erosive role of miR-17 in vivo. This effect involves the suppression of the IL-6 family autocrine amplifying loop through the direct targeting of JAK1 and STAT3 as shown in RA-FLS.FigureEffect of miR-17 lipoplex injection in Collagen induced arthritis (CIA) mice’s ankles.Disclosure of Interests:None declared