CFA-induced Arthritis Research Articles

Pharmacology and Articular Cartilage Regeneration: A New Strategy for Osteoarthritis

ABSTRACT Background: Osteoarthrosis is a slowly progressive, multifactorial, and non-inflammatory degenerative disease affecting synovial joints and is typically associated with aging or previous joint damage. Method: Fifty male Sprague Dawley rats were utilized in the study and divided into five groups, with ten rats in each group. The experiment was divided into two phases: in Phase 1, ten animals received 50 μL of saline in the left tibiotarsal joint and served as the control group (SAL), while the other four groups (n = 40) received a single dose of 50 μL complete Freund’s adjuvant (CFA) to induce monoarthritis. In Phase II, after the induction of monoarthritis, from the fourth week onward, the rats in the four CFA-induced monoarthritis groups were treated with 20 μl of gamma-amino butyric acid (GABA), serotonin (SERO), or a neurotransmitter cocktail (NTC) which were given intra-articularly once for three consecutive weeks, while one group was kept as an experimental control. The animals were monitored throughout the experiment, for ankle diameter, gait, and radiological assessment, and were then euthanized and subjected to histopathological evaluation at the conclusion of the study. Results: The study found that there were statistically significant differences in the degree of inflammation between the CFA group and the groups treated with GABA, SERO, and NTC. The CFA group exhibited the most severe and persistent inflammation, whereas the NTC group showed a milder and shorter inflammatory response than the control group (SAL). In terms of gait score, the NTC treatment resulted in a significant reduction in score compared to the treatments with GABA and SERO administered separately. The radiological scores revealed that NTC was the most effective treatment in mitigating radiographic evidence of joint damage, with a significant decrease in bone destruction. The histopathological examination showed that the NTC group had minimal tissue damage or inflammation and had a regenerative effect compared to the other treatment groups. Conclusions: In conclusion, the local administration of neurotransmitters was found to be effective in reversing inflammation, improving gait score, and mitigating radiological changes in CFA-induced arthritis in the murine model.

Therapeutic potential of hyaluronic acid hydrogel combined with bone marrow stem cells-conditioned medium on arthritic rats’ TMJs

Conditioned media (CM) is derived from mesenchymal stem cells (MSC) culture and contains biologically active components. CM is easy to handle and reduces inflammation while repairing injured joints. Combination therapy of the CM with cross-linked hyaluronic acid (HA) could ameliorate the beneficial effect of HA in treating degenerative changes of articulating surfaces associated with arthritic rats’ temporomandibular joints (TMJs). This study aimed to evaluate the therapeutic potential of HA hydrogel combined with bone marrow stem cells-conditioned medium (BMSCs-CM) on the articulating surfaces of TMJs associated with complete Freund’s adjuvant (CFA)-induced arthritis. Fifty female Sprague-Dawley rats were divided randomly into five equal groups. Rats of group I served as the negative controls and received intra-articular (IA) injections of 50 µl saline solution, whereas rats of group II were subjected to twice IA injections of 50 µg CFA in 50 µl; on day 1 of the experiment to induce persistent inflammation and on day 14 to induce arthritis. Rats of group III and IV were handled as group II and instead, they received an IA injection of 50 µl HA hydrogel and 50 µl of BMSCs-CM, respectively. Rats of group V were given combined IA injections of 50 µl HA hydrogel and BMSCs-CM. All rats were euthanized after the 4th week of inducing arthritis. The joints were processed for sectioning and histological staining using hematoxylin and eosin, Masson’s trichrome and toluidine blue special staining, and immunohistochemical staining for nuclear factor-kappa B (NF-κB). SPSS software was used to analyze the data and one-way analysis of variance followed by post-hoc Tukey statistical tests were used to test the statistical significance at 0.05 for alpha and 0.2 for beta. In the pooled BMSC-CM, 197.14 pg/ml of platelet-derived growth factor and 112.22 pg/ml of interleukin-10 were detected. Compared to TMJs of groups III and IV, TMJs of group V showed significant improvements (P = 0.001) in all parameters tested as the disc thickness was decreased (331.79 ± 0.73), the fibrocartilaginous layer was broadened (0.96 ± 0.04), and the amount of the trabecular bone was distinctive (19.35 ± 1.07). The mean values for the collagen amount were increased (12.29 ± 1.38) whereas the mean values for the NF-κB expression were decreased (0.62 ± 0.15). Combination therapy of HA hydrogel and BMSCs-CM is better than using HA hydrogel or BMSCs-CM, separately to repair degenerative changes in rats’ TMJs associated with CFA-induced arthritis.

Deciphering farnesol's anti-arthritic and immunomodulatory potential by targeting multiple pathways: a combination of network pharmacology guided exploration and experimental verification.

Farnesol (FAR), a sesquiterpene alcohol, has documented FAR's anti-inflammatory and antioxidant activities. Current study was undertaken to assess the efficacy and mechanism of FAR in arthritis by employing network pharmacology and experimental models. Two experimental models comprising formaldehyde- and complete Freund's adjuvant (CFA)-induced arthritis evaluated the efficacy of FAR in treating arthritis. Various parameters were assessed. Then, a network pharmacology approach was applied to gain further insight into the potential mechanism and signaling pathways. FAR significantly reduced paw volume and the arthritic score and improved the hematological and biochemical changes. Radiographic and histological examination showed the anti-arthritic efficacy of FAR, which was associated with down-regulation of pro-inflammatory mediators and upregulation of anti-inflammatory mediators. Network pharmacology analysis revealed that FAR may exert its anti-arthritic effects by targeting specific genes associated with arthritis. Pathway analysis revealed the involvement of three key signaling pathways (IL-17 signaling, TNF signaling, and toll-like receptor signaling) in the development and progression of arthritis. The results pointed out the protective attributes of farnesol against formaldehyde and CFA-induced arthritis via modulation of multiple targets. This study provides a valuable reference for the development of a new treatment or complementary therapy for arthritis.

Gingerol nanoparticles attenuate complete Freund adjuvant-induced arthritis in rats via targeting the RANKL/OPG signaling pathway.

Rheumatoid arthritis (RA) is characterized by inflammatory joint pathology leading to the degradation of articular bone and cartilage, primarily triggered by synovial inflammation, resulting in joint discomfort. The metacarpophalangeal and proximal interphalangeal joints are predominantly affected. Treatment typically involves a combination of biological and synthetic disease-modifying antirheumatic drugs (DAMARDs) alongside steroid therapy. The application of nanomedicine has been instrumental in enhancing treatment efficacy by facilitating controlled release of pharmacologically active compounds, thus augmenting bioavailability and enabling targeted drug delivery. Gingerol, a constituent of ginger, possesses multifaceted properties. including anti-inflammatory, anti-oxidant, antidiabetic, and antipyretic effects. In this study, gingerol-loaded poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs), coated with chitosan, were administered orally to rats over a period of 21days to address RA induced by complete Freund adjuvant (CFA). The rats were segregated into four experimental groups. Upon completion of the treatment regimen, blood samples were collected for the assessment of cyclooxygenase-2 (COX-2), RA factor, interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α). Subsequent gene expression analysis was conducted to evaluate the levels of interleukin-4 (IL-4), interleukin-17a (IL-17a), IL-6, interferon-gamma (INF-γ), TNF-α, interleukin-1 beta (IL-1β), osteoprotegerin (OPG), and receptor activator of nuclear factor kappa-B ligand (RANKL). Statistical analyses utilizing one-way ANOVA followed by Tukey tests were applied to the data. The gene expression profiling revealed significant disparities in mRNA levels of IL-1β, IL-6, IL-4, IL-17a, RANKL, INF-γ, and TNF-α between the CFA-induced arthritis group and the control group. Consequently, it was inferred that gingerol-loaded PLGA NPs coated with chitosan exhibited heightened therapeutic efficacy in addressing CFA-induced arthritis in rats.

Fixed oil from the Caatinga palm Syagrus coronata with effect on Complete Freund's Adjuvant (CFA)-induced arthritis.

Syagrus coronata, a native palm tree in the Caatinga domain, produces fixed oil (ScFO) used therapeutically and dietary by Northeast Brazilian communities. This study evaluated its anti-inflammatory potential of CFA-induced arthritis and its effect on behavioral parameters. In the acute model, ScFO at 25, 50, and 100mg/kg showed edematogenic effects similar to indomethacin at 4mg/kg (p > 0.05). In the arthritis model, 100mg/kg ScFO treatment was comparable to indomethacin (4mg/kg) (p > 0.05). TNF-α and IL-1β levels were significantly reduced in ScFO-treated groups at 25, 50, and 100mg/kg, and the indomethacin group (4mg/kg) versus the positive control (p > 0.05). Radiographs showed severe soft-tissue swelling and bone deformities in the control group, while the 100mg/kg ScFO group had few alterations, similar to the indomethacin group. Histopathological analysis revealed intense lymphocytic infiltration in the control group, mild diffuse lymphocytic infiltration in the indomethacin group, and mild lymphoplasmacytic infiltration with focal polymorphonuclear infiltrates in the 100mg/kg ScFO group. Behavioral analysis showed improved exploratory stimuli in ScFO and indomethacin-treated mice compared to the positive control (p > 0.05). ScFO demonstrated anti-inflammatory effects in both acute and chronic arthritis models, reducing edema and pro-inflammatory cytokines, and improved exploratory behavior due to its analgesic properties.

Mesua assamica (King & Prain) kosterm. bark ethanolic extract attenuates rheumatoid arthritis via down-regulating TLR4/NF-κB/COX-2/iNOS and activation of Nrf2/HO-1 pathways: A comprehensive study on in-vitro and in-vivo models

Ethnopharmacological relevanceRheumatoid arthritis (RA) is a multifactorial, polygenic inflammatory disease. Mesua assamica (King & Prain) Kosterm. (MA) is an endangered medicinal plant indigenous to South Asia, primarily to Assam in India. The tree bark is claimed to possess anti-inflammatory, anti-diabetic, anti-cancer, and anti-malarial properties; nevertheless, its role in RA has not been elucidated. Hence, this study aims to investigate the in-vitro and in-vivo anti-arthritic effects of Mesua assamica bark ethanolic extract (MAE). Aim of the studyThis study aims to investigate the anti-rheumatic potential of MAE in-vitro on RAW 264.7 cells for its anti-oxidant and anti-inflammatory activities and in-vivo on the CFA-induced adjuvant arthritis in the rat model. Materials and methodsWe investigated the possible therapeutic effects of MAE in-vitro using RAW 264.7 cells triggered by LPS. Meanwhile, adult Wistar rats were injected intradermally with 100 μl of CFA to induce arthritis, and they were given MAE orally at doses of 100 and 200 mg/kg for up to 28 days. Paw volume analysis, X-ray radiography, anti-oxidant levels analysis, gene and protein expression studies, and histological analysis were carried out to assess the effects of MAE in-vivo. ResultsMAE significantly mitigated the inflammation by reducing ROS levels and dropped the nitrite, PGE2, and COX-2 levels enhanced by LPS in-vitro. At the same time, MAE treatment reduced the paw and joint inflammation and increased the immune organ index in the CFA rats. Histopathology data revealed that MAE mitigated the CFA-induced lesions of the ankle joints and synovial tissues. Similarly, MAE significantly abated the secretion of pro-inflammatory cytokines, inhibited the protein expression of TLR4, NF-кB, COX-2, and iNOS, as well as improved the Nrf2 and HO-1 levels in-vitro and in-vivo. ConclusionAll the results highlighted the anti-rheumatic potential of MAE in RA in-vitro and in-vivo by inhibiting the TLR4/NF-кB/COX-2/iNOS and promoting the Nrf2/HO-1 signaling axis.

Possible Protective Effect of Trimetazidine Alone and in Combination with Methotrexate in Attenuating Joint Inflammation in Freund’s Adjuvants Induced Rheumatoid Arthritis in Rats

Abstract Rheumatoid Arthritis is a complex chronic auto-immune systemic illness. It is primarily characterized by synovial hyperplasia and localized pro-inflammatory cytokine production that causes chronic synovitis and a long-lasting inflammation of the joints and bones. The study's goal was to look into the potential anti-arthritic effects of trimetazidine in a rat model of Freund’s complete adjuvant (FCA)-induced arthritis, which has many clinical and biochemical similarities to rheumatoid arthritis (RA). The underlying antirheumatoid arthritis mechanism, as well as its impact on TLR4/NFKB pathway were investigated to understand and confirm its potential efficacy in RA treatment. Our study was based on 4 groups, control group received saline injections, trimetazidine (7.2 mg/kg/day) treated group, methotrexate (0.75 mg/kg/week) treated group and combination group that received both medications in the same doses for 21 days. The degree of ipsilateral paw swelling, ankle diameter, body weight, tissue expression levels of TLR4(toll like receptor) and NFKB (nuclear factor kappa b) in paw, and serum ACPA (anticitrullinated protein antibodies) were all measured. With a slight but not statistically significant impact on ankle diameter, trimetazidine and methotrexate considerably reduced the arthritic symptoms as indicated by ipsilateral paw volume. It had no discernible impact on body weight. Together with the overall improved synovial hyperplasia and cartilage disarrangements, TLR4 and NFKB expression in synovial tissue were reduced. In a rat model of CFA-induced arthritis, trimetazidine displays a considerable anti- inflammatory impact that is even better than methotrexate and has the potential to be anti-arthritic. Additionally, there is a large additive effect between the two medications.

Novel antiarthritic mechanisms of Azelaic acid against CFA-induced arthritis in rats by modulating pro- and anti-inflammatory cytokines network.

An immunologic system attacking the body's own tissues is a hallmark of autoimmune disorders, which encompass a wide range of unique conditions. Numerous essential biologic functions, including the regulation of the immune system, inflammation, cell division, and tissue repair, are carried out by cytokines. Natural compounds are an effective treatment for autoimmune illnesses by modulation of inflammatory cytokines and infiltration of leukocytes into the inflamed tissue. Here, anti-arthritic study was carried out using oral administration of Azelaic acid (AzA) for 28days with doses (20, 40, and 80mg/kg) in Complete Freund's Adjuvant (CFA) induced arthritis model. AzA ameliorated the adjuvant-induced arthritis by decreasing arthritic score, paw volume, improved body-weight alterations and serum levels of PGE2, 5-LOX and anti-ccp. AzA showed significant down regulation of NF-κB, COX-II, TNF-α, IL-17, IL-1β, IL-6, and up regulation of IL4 and IL10. Hemoglobin and RBCs count remarkably increased and ESR, CRP, platelets, WBCs levels markedly reduced in post treatment. In addition, the weakened SOD (superoxide dismutase), Catalase (CAT), Glutathione (GSH) activity and the increased levels of malondialdehyde (MDA) were all reversed by AzA treatment. And showed improved radiographical and histologic alterations in the structure of the joints. Molecular docking studies targeting COX-II, iNOS, TNF-α, 5-LOX, IL4, IL10, IL-6, and IL-17 establish a correlation between theoretical and experimental results. Results showed that AzA inhibit pro-inflammatory cytokines (COX-II, TNF-α, 5-LOX, IL-17, NF-κB, IL-1β, and IL-6) and increase anti-inflammatory cytokines, which supported the anti-arthritic and immunomodulatory potential of AzA.

Hemp seed oil ameliorates CFA-induced rheumatoid arthritis in rats by improving metabolic abnormalities and gut microflora

Hemp seed oil (HSO) acquired from hemp seeds (Cannabis sativa L.) is rich in polyunsaturated fatty acids and has been claimed for centuries to be beneficial for atopic dermatitis and inflammation. In this study, we investigated the beneficial effects of HSO against complete Freund’s adjuvant (CFA)-induced RA using 16S rRNA amplicon sequencing technology and metabolomics. Our findings demonstrated that HSO effectively alleviated rheumatoid arthritis symptoms such as paw swelling, synovial hyperplasia, joint inflammation, and bone destruction. Moreover, nuclear magnetic resonance (NMR)-based metabolomics revealed that fifteen potential biomarkers associated with RA in both serum and urine were restored to near normal levels by the administration of HSO. The 16S rRNA gene sequencing analysis revealed that the composition and number of the intestinal flora were restored to healthy levels in RA rats treated with HSO. These results suggested that HSO may attenuate RA by modulating gut microbiota composition and regulating metabolic abnormalities.

Arbutin attenuates CFA-induced arthritis by modulating expression levels of 5-LOX, NF‑κB, IL-17, PGE-2 and TNF-α.

Arbutin, a naturally soluble glycosylated phenol has antioxidant, antimicrobial, antitumor and anti-inflammatory properties. The current exploration appraises the treatment of arthritis by use of Arbutin (25, 50 and 100mg/kg) orally in CFA-induced rat arthritis model. Body weight changes, paw size, and joint diameter were recorded till the 28th day in the arthritic-induced rats. Hematological, biochemical, oxidative and inflammatory biomarkers were measured through the blood samples of anesthetized rats. Arbutin markedly decreased paw volume, PGE-2, anti-CCP and 5-LOX levels, however, maintained metabolic and hematological balance and prevented weight loss. Radiology and histology changes improved significantly in the ankle joints of rats. Moreover, Arbutin increased gene pointers such as IL-10 and IL-4 while significantly reducing the levels of CRP and WBCs, whereas Hb, platelets and RBCs count markedly raised in post-treatments. Antioxidant levels of SOD, CAT and GSH were improved and MDA level was reduced in treated groups. Rt-PCR investigation showed a significant reduction of the interleukin-1β, TNF-α, interleukin-6, cyclooxygenase-2, NF-κB and IL-17 and increased expression of gene pointers like IL-4, and IL-10 in treated groups. Assessment of molecular docking revealed a strong binding interaction of Arbutin against 5-LOX, IL-17, TNF-alpha and interleukin-6, cyclooxygenase-2, nuclear factor-κB, IL-4 and iNOS providing a strong association between experimental and theoretical results. As a result, Arbutin has significantly reduced CFA-induced arthritis by modulation of anti-inflammatory cytokines, i.e., IL-10 and IL-4, the pro-inflammatory cytokines panel such as NF-κB, TNF-alpha, IL-1β, IL-6, PGE-2, 5-LOX and COX-2 and oxidative biomarkers.

Ammodaucus Leucotrichus Seed Extract as a Potential Therapy in Animal Models of Rheumatoid Arthritis Induced by Complete Freund Adjuvant and Chicken Cartilage Collagen.

This study assessed the efficacy of an Ammodaucus leucotrichus seed extract to treat rheumatoid arthritis in rat models of this disease. Rheumatoid arthritis was induced in rats using two methods: immunization with 100 µL of Complete Freund Adjuvant (CFA) and immunization with 100 µL of a 3mg/ml solution of type II collagen (CII) from chicken cartilage. The therapeutic potential of the extract was assessed at different doses (150, 300, and 600mg/kg/day for 21 days in the CII-induced arthritis model and for 14 days in the CFA-induced arthritis model) and compared with methotrexate (MTX; 0.2mg/kg for the same periods), a commonly used drug for rheumatoid arthritis treatment in humans. In both models (CII-induced arthritis and CFA-induced arthritis), walking distance, step length, intra-step distance and footprint area were improved following treatment with the A. leucotrichus seed extract (all concentrations) and MTX compared with untreated animals. Both treatments increased the serum concentration of glutathione and reduced that of complement C3, malondialdehyde and myeloperoxidase. Radiographic data and histological analysis indicated that cartilage destruction was reduced already with the lowest dose of the extract (100mg/kg/dose) in both models. These results show the substantial antiarthritic potential of the A. leucotrichus seed extract, even at the lowest dose, suggesting that it may be a promising alternative therapy for rheumatoid arthritis and joint inflammation. They also emphasize its efficacy at various doses, providing impetus for more research on this extract as a potential therapeutic agent for arthritis.

Mechanistic Evaluation of Antiarthritic Effects of Eupatilin in CFA-Induced Arthritic Rats: An in Vivo and in Silico Study

Objectives In the present study, we explored the antiarthritic potential of Eupatilin on complete Freund's adjuvant (CFA)-induced arthritis rats. Materials and Methods In order to induce arthritis in the Sprague-Dawley rats, CFA vaccination was used, and Eupatilin was given to the rats at dose of 5, 10, and 20 mg/kg. Various biochemical and other parameters were studied to define the pharmacological benefit of Eupatilin. Results The study's findings revealed that Eupatilin increases rats’ body weight and significantly reduces hind paw volume. It also improves the antioxidant status of CFA rats by lowering malondialdehyde levels and increasing the concentration of endogenous antioxidants superoxide dismutase and GPx. Eupatilin also improved hematological markers, which were demonstrated to be altered in CFA rats as compared to the control. In the Eupatilin-treated group, the level of pro-inflammatory cytokines was found to be reduced as compared to CFA rats. In a molecular docking analysis, it showed favorable interaction with the active site of cyclooxygenase-2 (COX-2) by the formation of numerous nonbonded interactions by interacting with Cys32, Leu138, Pro 139, Val141, Ala142, and Cys145. It also showed a Vina score of −9.4 and interacts with the cavity volume of 4435 Å3. Conclusion Eupatilin showed favorable antiarthritic benefits in the CFA-induced arthritis model, as evidenced by reductions in paw volume, significant weight loss prevention, and maintenance of the healthy hematological and biochemical profile may be through inhibiting COX-2.

Manual acupuncture ameliorates inflammatory pain by upregulating adenosine A3 receptor in complete Freund’s adjuvant-induced arthritis rats

BackgroundAdenosine A3 receptor (A3R) exerts analgesic, anti-inflammatory, and anti-nociceptive effects. In this study, we determined the analgesic mechanism of manual acupuncture (MA) in rats with complete Freund's adjuvant (CFA)-induced arthritis and explored whether MA ameliorates inflammation in these rats by upregulating A3R. MethodsSixty Sprague Dawley (SD) rats were randomly divided into the following groups: Control, CFA, CFA + MA, CFA + sham MA, CFA + MA + DMSO, CFA + MA + IB-MECA, and CFA + MA + Reversine groups. The arthritis rat model was induced by injecting CFA into the left ankle joints. Thereafter, the rats were subjected to MA (ST36 acupoint) for 3 days. The clinical indicators paw withdrawal latency (PWL), paw withdrawal threshold (PWT), and open field test (OFT) were used to determine the analgesic effect of MA. In addition, to explore the effect of A3R on inflammation after subjecting arthritis rats to MA, IB-MECA (A3R agonist) and Reversine (A3R antagonist) were injected into ST36 before MA. ResultsMA ameliorated the pathological symptoms of CFA-induced arthritis, including the pain indicators PWL and PWT, number of rearing, total ambulatory distance, and activity trajectory. Furthermore, after MA, the mRNA and protein expression of A3R was upregulated in CFA-induced arthritis rats. In contrast, the protein levels of TNF-α, IL-1β, Rap1, and p-p65 were downregulated after MA. Interestingly, the A3R agonist and antagonist further downregulated and upregulated inflammatory cytokine expression, respectively, after MA. Furthermore, the A3R antagonist increased the degree of ankle swelling after MA. ConclusionMA can alleviate inflammatory pain by inhibiting the NF-κB signaling pathway via upregulating A3R expression of the superficial fascia of the ST36 acupoint site in CFA-induced arthritis rats.

Selegiline modulates inflammatory indicators in RAW 264.7 macrophages and LPS-aggravated CFA-induced rheumatoid arthritis in rats

Introduction and aim. Rheumatoid arthritis (RA) causes pain, inflammation, and deformities in numerous joints. Monoamine oxidase B (MOA-B) inhibitor selegiline exhibits anti-inflammatory characteristics and has the propensity to scavenge free radicals. Therefore, the aim of this research comprises of assessing the effect of selegiline on proinflammatory cytokines in RAW 264.7 macrophages as well as its capacity to improve various arthritic parameters in rats with lipopolysaccharide (LPS) accelerated complete Freund’s adjuvant (CFA) induced RA. Material and methods. In RAW 264.7 cells (lipopolysaccharide accelerated), nitric oxide (NO), tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6), inducible nitric oxide synthase (iNOS), and prostaglandin E2 (PGE2 ) were determined after treat ment with selegiline. Different arthritic parameters were analyzed after administration of selegiline in LPS accelerated CFA-induced arthritis in rats. Results. LPS escalates NO, TNF-α, IL-6, iNOS, and PGE2 quantities in the RAW 264.7 cells, which was minimized by selegiline at 100 µg/mL and 150 µg/mL respectively. In rats, CFA induction causes a decrease in body weight, elevation of paw volume, splenic index, and arthritic index, which are further accelerated by LPS. 20 mg/kg of selegiline managed all these arthritic parameters effectively, including TNF-α, IL-6, and a few other biochemical parameters. Conclusion. Selegiline may be beneficial in RA extenuating joint and cartilage damage, and modulating inflammatory responses.

Estragole Ameliorates CFA Induced Rheumatoid Arthritis Symptoms in Wistar Rats by Inhibiting JAK-2/STAT-3 Pathway.

The present study was conducted to scrutinize the pharmacological effect of Estragole (ESG) against CFA-induced arthritis in rats. The rats underwent induction of arthritis using the administration of CFA and after that, the rats were randomly divided into five different groups, where three groups correspond to diverse dosages of ESG, and the other two were control and CFA-arthritic control. Results of the study suggested that ESG in a dose-dependent manner, improves body weight and arthritis score of rats as evidenced by reduction of hind-paw volume. ESG also improved the antioxidant status of rats by reducing MDA levels and enhancing the concentration of endogenous antioxidants SOD and GPx. The level of pro-inflammatory cytokines was also found to be reduced in the case of ESG treated group as compared to CFA-group. In a western blot analysis, ESH showed downregulation of p-JAK-2/STAT-3. The study provided concrete evidence for the protective effect of ESG against rheumatoid arthritis in rats.

Antiarthritic and Antioxidant Activities of Antrocaryon micraster Seed Extract and Its Fractions.

Rheumatoid arthritis (RA) is an incurable debilitating disease which attacks the joints and impairs quality of life. Antrocaryon micraster is used to treat RA in African traditional medicine. However, its antiarthritic activity has not been pharmacologically studied. This study, therefore, reports the antiarthritic and antioxidant activities of A. micraster seed extract and its fractions. The seed extract (ASE) was produced by Soxhlet extraction and partitioned into petroleum ether (ASEP), ethyl acetate (ASEE), and aqueous (ASEA) fractions. The total polyphenolic content, DPPH antioxidant activity, and in vitro arthritic activity using the protein denaturation assay were evaluated for ASE and its fractions. The arthritic activity of the crude extract (ASE) and its most effective fraction (ASEA), in the in vitro assay, were then evaluated against CFA-induced arthritis in rats. The polyphenolic constituent of ASE was estimated to be 13.00 ± 0.00 mg/100 mg of GAE. ASEA contained the highest quantity of polyphenolic constituents (10.76 ± 0.00 mg/100 mg of GAE) among the fractions of the extract. ASE and ASEA produced profound antioxidant activity (IC50 = 20.17 ± 1.291 and 19.35 ± 0.865, respectively) which were similar to that of ascorbic acid (IC50 = 17.35 ± 0.500) in the DPPH free radical scavenging assay. Furthermore, in vitro antiarthritic activity of ASEA was 13.63 and 5.75 times higher than the antiarthritic activity of the crude extract and diclofenac sodium, respectively. In the CFA-induced arthritis assay, both ASE and ASEA significantly (P < 0.001) inhibited cachexia, paw edema, infiltration of inflammatory cells, pannus formation, and synovium damage. These results indicate that A. micraster seed extract and its fractions possessed significant antiarthritic activity via inhibition of oxidative stress, inflammation, protein denaturation, infiltration of inflammatory cells, and synovium injury due to its constituents such as polyphenols and phytosterols.

Investigating the anti-inflammatory and anti-arthritis effects of fucoidan from a brown seaweed

Severe inflammation in joints caused by the detrimental effects of the immune system is termed Rheumatoid arthritis. The unconstrained proliferation of immune cells and pro-inflammatory cytokines deteriorates Synovium which secretes synovial fluid to lubricate joints and cartilage. Non-steroidal anti-inflammatory drugs (NSAIDs) are the only therapeutics for treating rheumatoid arthritis, and long-term intake causes serious side effects on the organs. Fucoidan, a sulfated polysaccharide found on the cell walls of brown algae shows bioactive potential. In our study, fucoidan was extracted from Padina pavonica (PD), Stoechospermum marginatum (StM), Spatolossum macrodontum (SpM), Dictyota bartayresiana (DD), and Turbinaria decurrens (TD) and evaluated for anti-inflammatory and anti-arthritis activities. Fucoidan was extracted and evaluated for anti-inflammatory activity in vitro using RAW 264.7 macrophage cell lines, followed by in vivo anti-arthritis activity on Wistar male rats. Nitric oxide suppression was comparatively high in fucoidan from TD (IC50 − 12.93 µg/mL). Purified fucoidan from TD, significantly reduced inflammation, size of paw edema, downregulated proinflammatory cytokines (IL-6, IL-1β, TNF-α), and upregulated anti-inflammatory cytokine (IL10) in CFA-induced arthritis in Wistar male rats. Biochemical parameters like SOD, CAT, GSH, GPX, and GST and haematological parameters like total-protein, albumin, haemoglobin, and RBC were upregulated, and other parameters like urea, uric acid, creatinine, bilirubin, SGOT, SGPT, ALP, WBC, ESR, RF, and CRP were downregulated. Histopathology of the liver, kidney, and ankle joints reveals that fucoidan intake restrained inflammation and tissue damage. Therefore, fucoidan extracted from TD is a potential candidate for the treatment of rheumatoid arthritis.

Navel orange peel ethanolic extract and naringin ameliorate CFA-induced arthritis in Wistar rats through their modulatory effects on Th1/Th2/Th17 cytokines and oxidative stress.

Rheumatoid arthritis (RA) is an autoimmune illness affecting joint articulations, leading to a disability state. Currently, there is no satisfying optimal therapy except for immunosuppressants, which have variable and bad effects after long-term use. Hence, researchers have attempted to develop other alternative, safer, and more effective natural treatment agents that are effective and without undesirable effects. The objective of this research is to assess the antiarthritic properties of navel orange peel ethanolic extract (NOPEE) and naringin (NAR) in experimentally induced RA in male Wistar rats. RA was induced via two successive subcutaneous injections of 0.1 mL complete Freund's adjuvant (CFA) into a footpad of the right hind leg. The arthritic rats were orally treated with 100 mg/kg body weight (b.w.)/day of NOPEE or with 25 mg/kg b.w./day of NAR for 14 days. Results showed that treatment with NOPEE or NAR obviously counteracted the increased ankle joint circumference, inflammatory cell infiltration, pannus development, cartilage degradation, and synovial hyperplasia that developed in CFA-induced arthritic rats. Additionally, the elevation of serum rheumatoid factor (RF), prostaglandin E2 (PGE-2), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-17 (IL-17) were significantly declined in parallel to enhanced level of serum interleukin-4 (IL-4). Furthermore, NOPEE and NAR supplementation, reversed the negative oxidative effects of lipid peroxidation (LPO), nitric oxide (NO), as well as improved the antioxidant glutathione level (GSH), glutathione reductase (GR) and superoxide dismutase (SOD) activities. Overall, the anti-arthritic effects of NOPEE and NAR may be mediated through their modulatory effects on T helper (Th)1/Th2/Th17 cytokines, oxidative stress, and the antioxidant defense system.

Immunostimulant, hepatoprotective, and nephroprotective potential of Bacillus subtilis (NMCC-path-14) in comparison to dexamethasone in alleviating CFA-induced arthritis.

To investigate and compare efficacy as well as safety of Bacillus subtilis and dexamethasone (Dexa) in complete Freund's adjuvant (CFA)-induced arthritis, we used glucocorticoid monotherapy (Dexa 5mg/kg/day) and B. subtilis (1 × 108CFU/animal/day p.o) as pre-treatment and concurrent treatment for a duration of 35days. Specific emphasis was on chronic aspect of this study since long-term use of Dexa is known to produce undesirable side effects. Treatment with Dexa significantly attenuated the arthritic symptoms but produced severe side effects like weight loss, increased mortality, immunosuppression, and altered histology of liver, kidney, and spleen. Oxidative stress was also elevated by Dexa in these organs which contributed to the damage. Treatment with B. subtilis improved symptoms of arthritis without producing any deleterious side effects as seen with Dexa therapy. Immunohistochemistry (IHC) profile revealed decreased expression of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), interleukin (IL)-1β, tumor necrosis factor alpha (TNF-α), and increased nuclear factor erythroid 2-related factor 2 (Nrf-2) expression by B. subtilis and Dexa treatment in ankle joint of arthritic mice. Radiological scores were also improved by both treatments. This study concludes that B. subtilis could be an effective alternative for treating arthritis than Dexa since it does not produce life-threatening side effects on prolong treatment.

Attenuation of CFA-induced arthritis through regulation of inflammatory cytokines and antioxidant mechanisms by Solanum nigrum L. leaves extracts.

Solanum nigrum L. is a popular traditional medicine for various inflammatory conditions including rheumatism and joint pain. The current study aimed to evaluate the anti-arthritic mechanism of Solanum nigrum L. Four extracts were prepared using n-hexane, methanol, chloroform, and water. The anti-nociceptive and anti-inflammatory activity was carried out with 100, 200, and 300mg/kg body wt. PO of each extract by the hot plate and carrageenan-induced paw oedema methods, respectively. The anti-arthritic study was performed with chloroform and aqueous extracts (300mg/kg) in complete Freund's adjuvant (CFA)-induced arthritis. Paw size (mm), ankle joint diameter (mm), and latency time (sec) were recorded on day 0 and every 4th day till 28days. The hematological, inflammatory, and oxidative biomarkers were estimated. Results showed that significant analgesia (p < 0.05) and reduction in paw inflammation were achieved with all extracts. The highest percent inhibition in Carrageenan-induced inflammation was achieved with 300mg/kg of chloroform (72.19%) and aqueous (71.30%) extracts, respectively. In the CFA model, both extracts showed a significant reduction in paw size and ankle joint diameter (p < 0.05). The RT-qPCR analysis revealed the upregulation of interleukin-4 and interleukin-10, and down-expression of interleukin-1β, interleukin-6, tumor necrosis factor-α, cycloxygenase-2, nuclear factor-κB, prostaglandin E synthase 2, and interferon-γ. A significant increase in superoxide dismutase, catalase, and glutathione levels was observed. Hence, it is concluded that Solanum nigrum L. leaf extracts regulate the expression of inflammatory markers and improve oxidative stress resulting in the attenuation of CFA-induced arthritis.