Adjuvant-induced Rheumatoid Arthritis Research Articles

Antirheumatoid Arthritic Effects of Sabia parviflora Wall. Leaf Extracts via the NF-κB Pathway and Transient Receptor Potential Protein Family.

As an important traditional medicine of Buyi and Miao ethnic groups in Guizhou, Sabia parviflora Wall. provides antiviral properties against hepatitis, eliminates wind and dampness, and exhibits anti-inflammatory and pain relief properties. It has also been shown to treat rheumatoid arthritis (RA) and other diseases. However, the pharmacodynamic mechanism of S. parviflora Wall. for RA has not been reported. In this study, we identified the effective compounds of S. parviflora Wall. leaves against RA and discussed the mechanism against complete Freund’s adjuvant-induced arthritis (AIA) based on inflammatory proteins and transient receptor potential (TRP) proteins. S. parviflora Wall. leaf extracts (0.64 g/kg, 0.32 g/kg, and 0.16 g/kg, once daily) were given orally for 21 days. On the 15th day of complete Freund’s adjuvant-induced RA, the effects of this medicine on RA rats were investigated. S. parviflora Wall. extracts increased body weight, decreased foot swelling, and reduced thymus and spleen indices in model rats. Most of pannus in the synovial tissue of RA rats disappeared upon treatment, and the local inflammatory cells were greatly reduced when given the fraction of n-butanol (0.64 g/kg/d, 0.32 g/kg/d, and 0.16 g/kg/d) of 70% alcohol-soluble fraction of S. parviflora Wall. leaves. In addition, the release of inflammatory factors such as tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-10 (IL-10), interleukin-15 (IL-15), and vascular endothelial growth factor (VEGF) in the RA rat serum was inhibited. The active compounds inhibited the expression of TNF-α, IL-1β, IL-6, IL-10, IL-15 and nuclear factor kappa-Bp65 (NF-κBp65) inflammatory protein and TRP protein transient receptor potential melastatin-5 (TRPM-5) and transient receptor potential channel-6 (TRPC-6), to reduce the expression of VEGF in synovial tissue of RA rats and relieve redness and edema. High-performance liquid chromatography identified six flavonoids and three triterpenoid saponins as active compounds. These findings suggest S. parviflora Wall. leaves may play a role in RA treatment by inhibiting the release of inflammatory factors as well as participating in the inflammatory protein expression in the NF-κB pathway and TRP protein family.

The Effect of Chia Seed Extracts against Complete Freund’s Adjuvant-Induced Rheumatoid Arthritis in Rats

Background and Objectives. It is known that the oxidation of chia seeds is minimal or absent, due to the presence of bioactive compounds, having a great potential in the foods and pharmacological industry. This investigation was done to estimate the nutrition values and anti-oxidant activity of chia seed extracts. Materials and Methods. The protective effects of chia seed extract against complete Freund’s adjuvant (CFA) rheumatoid arthritis in rats were investigated with 100, 200, 300, and 400 ppm/kg BW rat/day chia seeds aqueous extract for 4 weeks. Results. The results obtained revealed that chia seed extract contained high proportions of protein and total dietary fiber values of 21.35% and 27.24%, respectively. Moreover, the results reported that the mineral contents were the greatest in phosphorous, potassium, magnesium, calcium, and sodium (450.09, 410.46, 245.22, 218.69, and 120.34 mg/100 g, respectively). Meanwhile, iron, zinc, and copper were the lowest in chia seeds (9.26, 4.67, and 3.66 mg/100 g, respectively). Besides, vitamins C and E reported 1.65 and 0.82 mg/100 g, respectively. Chia seed extracts were effective in vitro for the bioactive components such as phenolics, flavonoids, and anti-oxidant activities. The biomarkers of complete blood picture, lipid profile, anti-oxidant enzymes, TNF-α, and IL-10 had been improved. Histopathological examination of the rat knee confirmed health amelioration, revealing that chia seed extract consumption can lower pathological changes in injured rheumatoid arthritis rats. Conclusion. It could be seen that the chia seed extracts alleviated the harmful effect of rheumatoid arthritis CFA-induced rats.

Effects of Pereskia aculeate Miller Petroleum Ether Extract on Complete Freund's Adjuvant-Induced Rheumatoid Arthritis in Rats and its Potential Molecular Mechanisms.

Objective: To investigate the therapeutic effect of petroleum ether extract of P. aculeate Miller (PEEP) on rheumatoid arthritis (RA). Methods: In vitro: The Cell Counting Kit-8 (CCK-8) was used to detect cell activity and select the optimal concentration of the extract; the effective site was screened by nitric oxide (NO) colorimetric method and Q-PCR method; the expression of p38, p-p38, p-MK2, and Tristetraprolin (TTP) in RAW 264.7 cells were detected by Western blot. In vivo: The rat model was established by complete Freund’s adjuvant (CFA). The different doses of PEEP on CFA rats were observed with life status, paw swelling, spleen index, X-ray, Hematoxylin eosin (HE) staining; the secretion of Tumor necrosis factor α (TNF-α), interleukin-6 (IL-6) and Prostaglandin E2 (PGE2) were detected by Enzyme linked immunosorbent assay (ELISA); the expressions of p38, p-p38, p-MK2, and TTP in the ankle joints of CFA rats were detected by Western blot. Result: In vitro: PEEP, Ethyl Acetate Extract of P. aculeate Miller (EEEP), N-butanol Extract of P. aculeate Miller (BEEP) have no toxic effects on RAW264.7 macrophages. PEEP, EEEP, and BEEP reduce the secretion of NO in RAW264.7 cells induced by lipopolysaccharide (LPS), only PEEP significantly inhibited the mRNA expression levels of inflammatory factors TNF-α and IL-6; PEEP-dependently reduce the secretion of TNF-α and IL-6, decrease the expression of p-p38 and p-MK2, and the level of TTP phosphorylation in LPS-induced RAW264.7 cells. In vivo: PEEP improve the living conditions of CFA rats, reduce foot swelling, spleen index, bone surface erosion and joint space narrowing; reduce the formation of synovial cells, inflammatory cells and pannus in the foot and ankle joints. PEEP reduce the secretion of TNF-α, IL-6, PGE2 in rat serum, downregulate the expression of p-p38 and p-MK2 in the ankle joint, and reduce the phosphorylation of TTP. Conclusion: PEEP improve the living conditions of CFA rats, reduce the degree of foot swelling, protect immune organs, reduce inflammatory cell infiltration, cartilage damage, pannus formation, reduce inflammation and RA damage. The mechanism through regulating the signal pathway of p38 mitogen-activated protein kinase (p38/MAPK), which reduces the release of TNF-α, IL-6, and PGE2 in the serum.

Pharmacological effect of Lidang Tang against adjuvant-induced rheumatoid arthritis in rats

Rheumatoid arthritis (RA) is a great concern across the globe. It is characterized as an autoimmune disease, where the body's immune system mistakenly attacks bone joints causing them to erode which leads to deformity in structure. Thus, the present study was conducted to elucidate the effect of Lidang Tang (LDT) decoction, a traditional Chinese medicine against rheumatoid arthritis. The rheumatoid arthritis was induced in Sprague-Dawley albino rats by intradermal injection of bovine collagen II type at the tail. Results suggested that LDT reduces paw swelling and arthritic scores in rats. The protective effect of LDT against RA was further substantiated by histopathological analysis of synovial tissue of rats, where LDT improves the architecture of the tissues as compared to CIA rats. It also reduces oxidative stress and inflammation in CIA rats as compared to the disease model group. The serum level of anti-collagen II-specific immunoglobulins (IgG1 and IgG2a) was reduced significantly in LDT treated group. In a western blot analysis, LDT treated group showed a significant reduction in the expression of NF-ĸB and COX-2 as compared to CIA rats. Collectively, our study for the first time demonstrated the protective effect of Lidang Tang decoction against rheumatoid arthritis.

Structure of type II collagen from sturgeon cartilage and its effect on adjuvant-induced rheumatoid arthritis in rats.

The purpose of this paper was to extract and characterize type II collagen of sturgeon cartilage (SC-CII), and to explore the effects of taking SC-CII orally on rheumatoid arthritis (RA) in rats. SC-CII showed a triple-helix structure (RPN = 0.12), with d1 of 11.82 Å and d2 of 4.08 Å, which was analyzed by FT-IR, CD, XRD, and MS. It was constructed of the repeating tripeptide unit Gly-X-Y, where X and Y are generally Pro or Hyp, proved by amino acid composition and peptide mass fingerprinting. Furthermore, the effects of SC-CII on RA were evaluated. Ankle thickness was significantly decreased in SC-CII groups, with changes in lymphocyte proliferation also observed. Compared with the model control group, there was an evident decrease in TNF-α, IL-1β, COX-2, MCP-1, and TLR-4 mRNA levels, but no remarkable differences in APF, MMP-3, and MyD88 mRNA levels in the SC-CII groups. In addition, TNF-α, IL-1β, RF, Anti-CII Ab were significantly reduced in the SC-CII groups, proved by ELISA. Therefore, SC-CII showed alleviating effects on RA through the TLR4/MyD88-NFκB pathway.

Comparative study of cannabinoid receptor 2 agonist and dexamethasone in experimentally induced rheumatoid arthritis

To assess the impact of cannabinoid receptor 2-agonist (JWH-133) vs. dexamethasone in rats in a model of adjuvant-induced rheumatoid arthritis. The study utilized male albino rats weighing between 180 and 200 gm. The obtained data from the present study used dexamethasone orally daily for one 1week (1.5mg/kg), Cannabinoid receptor 2 agonists (JWH133) was administered as the other medication, at a dose of 4mg/kg I.P. injection daily for one week. Normal rats that received saline and DMSO showed i

Integrative serum metabolomic analysis for preventive effects of Yaobitong capsule in adjuvant-induced rheumatoid arthritis rat based on RP/HILIC-UHPLC-Q-TOF MS

Yaobitong capsule (YBTC) has been used for the prevention and treatment of inflammation-related lumbago and leg pain. However, its intervention mechanism still remains unclear. This study was aimed to evaluate the control efficiency of YBTC on adjuvant-induced rheumatoid arthritis (RA) rats by metabonomic method and to explore its possible anti-arthritis mechanism. Taking into account the complexity of endogenous metabolites in serum samples, an integrated metabolomics method based on RP/HILIC-UHPLC-Q-TOF MS was developed, to overcome the limitations of a single chromatographic in this study. The results showed that 32 potential biomarkers of arthritis were identified, primarily related to amino acid metabolism, glucose metabolism, lipid metabolism and nucleotide metabolism. Further receiver operating characteristic analysis revealed that the area under the curve of two down-regulated metabolites (3-Hydroxy-hexadecanoic acid, 2-Oxoarginine) and one up-regulated metabolite (l-Glutamic acid) among 32 biomarkers were 0.906, 0.969 and 1.000, respectively, indicating that high predictive ability of this method for RA. In this study, an integrated serum metabolomics method based on high-resolution mass spectrometry was successfully established for the first time to study the intervention mechanism of YBTC in RA, providing evidence regarding the clinical application of YBTC and a new insight for the prevention of RA in the future.

The role of the metabolic status of lymphocytes in the pathogenesis of rheumatoid arthritis

Objective: to study in dynamics metabolic changes in lymphocytes on the model of adjuvant-induced rheumatoid arthritis (RA) in warm-blooded animals.Material and methods. The RA model was elicited by administering Freund's complete adjuvant (AF) to male Wistar rats. Lymphocyte metabolism was corrected with a mixture of citric and succinic acids, which was injected for 4 weeks (at doses of 17 and 88 mg/kg body weight) from the first day of model formation. Animals of the control group were injected subcutaneously with isotonic sodium chloride solution. The total number of leukocytes, the size of lymphocytes and the activity of lactate dehydrogenase (LDH) and succinate dehydrogenase (SDH) in these cells were determined. Changes in the musculoskeletal system were assessed radiographically and actometrically in the «open field» model.Results and discussion. In the first 2 weeks of the experiment in the model group, an increase in the level of leukocytes by 65% compared to the control (p=0.002) was noted, an increase in the activity of lymphocytes SDH by 51% and a decrease in the total («horizontal») mobility of animals by 30% were also revealed. Subsequently, the level of leukocytes decreased by 25%, the activity of LDH and SDH – by 38%, the radius of lymphocytes – by 14% (p<0.01). The action of carboxylic acids was dose-dependent: at the maximum dose, no statistically significant differences in the total number of leukocytes, LDH and SDH activity in lymphocytes, as well as their sizes were found between the group with adjuvant- induced RA and the control group.Conclusion. In animals with adjuvant-induced RA, correction of the metabolic status of lymphocytes with a mixture of citric and succinic acids, which are key substrates of the Krebs cycle, led to a decrease in structural damage of musculoskeletal system and, as a consequence, to the maintenance of normal range of motion.

Adverse effect of rheumatoid arthritis on male Wistar rat’s fertility: protective role of Costus extract

Rheumatoid arthritis (RA) is a systemic autoimmune complaint. Advanced treatments resort to the traditional herbal therapy. The aim of this study is to assess the protective effect of Costus extract on the fertility of male rats with Freund's adjuvant-induced rheumatoid arthritis. Thirty male adult Wistar rats (190-200 g) were divided into six groups. They were subdivided into three groups; group I was the control group that received distilled water, and groups II and III received two various doses of Costus extract (200 and 400 mg/kg, respectively) for 60 days. Another three groups were subjected to RA induction via Freund's adjuvant. Rats were injected a dose of 0.1 ml of Freund's complete adjuvant (FCA) in the planter area of the left hind paw and then subdivided into 3 groups. Group I of RA-induced rats were given distilled water. The other two groups were given orally (200 and 400 mg/kg dosage of extract, respectively) from the 2nd day of RA induction for 60 days. Sex organ relative weight, sperm concentration assay, testicular histopathology and immunohistochemistry of androgen receptors, TNF α, and BAX protein were determined. The results showed that RA caused a significant decrease in the relative weight of sex organs and sperm count, which were relatively improved by doses of Costus (200, 400 mg/kg). RA induction caused testicular degeneration which markedly enhanced with Costus treatment as shown in histopathological sections. RA caused a reduction in %IHC of androgen receptors and increased expression level of both TNF α and BAX protein. Using IHC, it was revealed that RA caused a reduction in the expression level of androgen receptors and an increase in the expression of both TNF α and BAX protein. We can conclude that Costus speciosus had a potentially valuable role in improving fertility disorders caused by RA.

Cimiracemate A confers protection on arthritic neonatal rats via regulation of iNOS/NF-κB/TLR-4 pathway

Purpose: To investigate the protective effect of cimiracemate A on Freund’s adjuvant-induced rheumatoid arthritis (RA) in neonatal rats, and the underlying mechanism.
 Methods: Rheumatoid arthritis was induced in rat pups using Complete Freund’s adjuvant (100 µg/100 µL/body weight) which was intra-dermally injected at the tail region. After 21 days of establishment of RA, the rats were randomly assigned to four groups of ten rats each: control group, RA group, 5 mg/kg cimiracemate A group, and 10 mg/kg cimiracemate A group. Cimiracemate A was orally administered for 45 days. The effect of cimiracemate A on oxidative stress biomarkers, superoxide dismutase (SOD), malondialdehyde (MDA) and reduced glutathione (GSH) were determined using standard methods. Plasma levels of the inflammatory cytokines interleukin 1β (IL-1β) and tumor necrosis factor-α (TNF-α), and prostaglandin E2 (PGE-2) and matrix metalloproteinase-3 (MMP-3) were determined using enzyme-linked immunosorbent assay (ELISA). Western blotting was used to determine the levels of protein expressions of iNOS, NF-κB and TLR-4.
 Results: The level of MDA significantly increased and the level of GSH significantly decreased in RA group relative to control group (p < 0.05) following treatment with cimiracemate A. SOD activity was significantly reduced in RA group, when compared with control group (p < 0.05). However, treatment with cimiracemate A significantly and dose-dependently reversed the altered levels of MDA and GSH and SOD activity, when compared with RA group (p < 0.05). Plasma levels of IL-1β, TNF-α, PGE-2 and MMP-3 were significantly higher in RA group than in control group, but were significantly and dosedependently reduced after treatment with cimiracemate A (p < 0.05). There were significant increases in the levels of expression of iNOS, NF-κB and TLR-4 proteins in the chondrocytes of RA group, relative to control group (p < 0.05). However, treatment with cimiracemate A significantly and dose-dependently down-regulated the expressions of these proteins, when compared with RA group (p < 0.05).
 Conclusion: The results of this study indicate that cimiracemate A confers some degree of protection on arthritic neonatal rats via a mechanism that involves regulation of iNOS/NF-κB/TLR-4 pathway.

Beneficial anti-inflammatory and anti-arthritic activities of Grape seed proanthocyanidin extract in Freund's Complete Adjuvant-induced Rheumatoid arthritis in rats

Rheumatoid arthritis (RA) is systemic autoimmune inflammatory disorder withpermanent joint destruction. The Grape seed proanthocyanidin extract (GSPE) beneficial effects were evaluated in adjuvant-induced arthritis (AIA). RA was induced by 0.1 mL Freund's complete adjuvant (CFA) that was injected once into the rat's right hind leg footpads. Thirty male rats were equally divided into 5 groups. Group I (Normal control): Rats received no drugs, Group II (CFA- induced arthritis for treatment): Rats injected with 0.1 ml CFA on the 1st day and sacrificed after 29 days of CFA injection, Group III (CFA + GSPE treated): After 14 days of CFA injection, rats treated with GSPE (100 mg.kg-1 b wt./day) for 15 sequential days. Group IV (CFA- induced arthritis for protection): Rats injected with 0.1 ml CFA on 15th day of the experiment and sacrificed 15 days later after arthritis induction. Group V (CFA + GSPE protected): Rats pretreated with GSPE (100 mg.kg-1 b wt./day) one day before CFA injection and 14 days later after arthritis induction. A significant up-regulation in IL-1β, MMP‐3 and LOX-1 gene expressions were observed in the arthritic rats' synovial membrane. However, significant down-regulations in all previous parameters were observed in GSPE treated groups. Also, GSPE significantly activated Nrf2 gene in liver with marked up-regulation of Lactoferrin (LF) gene in arthritic rats' synovial membrane. Interestingly, improvement of histopathological alterations in synovium and stifle joint capsule of arthritic rats supported the anti-arthritic effects of GSPE. Conclusively, GSPE ameliorates adjuvant induced arthritis via its anti-arthritic and joint protecting properties.

Total Saponins from Nigella glandulifera Seeds Ameliorate Adjuvant-Induced Rheumatoid Arthritis in Rats by Inhibition of an Inflammatory Response and Bone Erosion.

Rheumatoid arthritis (RA) is a widespread inflammatory disease whose clinical manifestations are joint swelling, pain, and disability, affecting approximately 1% of individuals worldwide. Conventional anti-RA drugs currently used in clinic have severe side effects. The present study is aimed at investigating the antiarthritic effects of total saponins from Nigella glandulifera seeds (TSNGS) in rats with adjuvant-induced rheumatoid arthritis (AIA). Arthritis score, paw swelling, and body weight were monitored throughout the period of TSNGS treatment. The histopathological features and levels of cytokines, including IFN-γ, TNF-α, IL-1β, IL-4, IL-6, IL-10, and IL-17A, and OPG/RANKL signaling, were measured to determine the amelioration by TSNGS and its potential mechanisms on the inflammatory response and bone erosion. The differentiation of regulatory T cells (Tregs) in serum was assessed by flow cytometry. The results demonstrate that TSNGS at 10 mg/kg, 50 mg/kg, and 250 mg/kg inhibited AIA-induced clinical score, paw swelling, and histological changes. TSNGS reduced the immune-inflammatory reaction by restoring the secretion and expression of inflammatory cytokines and elevating the proportion of CD4+ CD25+ Tregs, accompanied by an increase in transcription factor Foxp3 levels. TSNGS also displayed bone protection by upregulation of the OPG/RANKL pathway. Collectively, TSNGS inhibited arthritis in AIA rats and so represents a potential novel treatment for RA.

Antiarthritic activity of asiaticoside against the Freund's complete adjuvant-induced rheumatoid arthritis in experimental wistar rats

Background: Rheumatoid synovial inflammation occurs at the synovial joint, which is activated by the articular system and mediated by the immune cells. In the case of untreated rheumatoid arthritis, the inflammation may affect the extra-articular system including cardiovascular, hepatic, respiratory, circulatory, and neuromuscular systems, which may ultimately interfere with the quality of life of patients with rheumatoid arthritis. This, in turn, reduces the life expectancy of the patient. Materials and Methods: The effectiveness of asiaticoside was evaluated by the Freund's complete adjuvant (FCA)-induced rheumatoid arthritis model using experimental Wistar rats. According to our results, asiaticoside exhibited potent anti-arthritic activity in contrast to the control animals in the FCA-induced rheumatoid arthritic model. Results: The parameters of arthritis such as reduced body weight, increased paw volume and pro-inflammatory cytokines, and cartilage destruction were significantly (P < 0.05) affected by the administration of asiaticoside. It significantly (P < 0.01) increased the body weight, decreased the paw volume and pro-inflammatory cytokines, restored the blood components, increased the levels of antioxidant enzymes, and protected the cartilage. Conclusion: Asiaticoside exhibited potential anti-arthritic activity against FCA-induced rheumatoid arthritis in experimental rats.

Zingerone (4-(four-hydroxy-3-methylphenyl) butane-two-1) modulates adjuvant-induced rheumatoid arthritis by regulating inflammatory cytokines and antioxidants

ABSTRACT Objective Ginger (Zingiber officinale Roscoe) is considered to be one of the most commonly consumed dietary condiments of the world. The present study was designed to explicate the protective role of zingerone; an active ingredient of ginger in complete Freund’s adjuvant (FCA)-immunized arthritic rats. Methods 24 Wistar rats were divided into 4 groups with 6 rats each. Group I as control followed by group II, III and IV were treated with single intradermal injection of FCA (0.1 ml = 100 µg) to induce rheumatoid arthritis. Group III and IV were also administered with zingerone orally at 25 mg/kg b.w for 3 weeks at two different time points. Results Adjuvant-treated rats exhibited a significant increase in lipid peroxidation and a reduction in the enzymatic antioxidants such as SOD, catalase and GPx, in the liver and joint tissues. Moreover, FCA inoculation resulted in the increase in levels of NF-κB, TGF-β, TNF-α, IL-1β, IL-6 and Hs-CRP and a decrease in IL-10 levels. Zingerone significantly reduced the levels of NF-κB, TGF-β, TNF-α, IL-1β, IL-6 and Hs-CRP and markedly increased IL-10 levels. Levels of antioxidant enzymes were also restored by zingerone treatment. Discussion Oral administration of zingerone ameliorated inflammatory outburst and decreased oxidative stress, suggesting its role in the prevention of rheumatoid arthritis. Further mechanistic insights are necessary to study the exact mechanism involved.

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.

A molecular docking study: Cepharanthine protects articular cartilage against arthritis by Wnt/PI3K/TLR-3 signaling

Rheumatoid arthritis (RA) is an autoimmune disorder that causes joint inflammation and permanent disability in affected patients. This study investigated the protective effect of cepharanthine treatment against Freund's adjuvant-induced RA in a rat model. RA was induced in rats by injection of incomplete Freund’s adjuvant (IFA) into the plantar surface on days 0 and 9. Cepharanthine at a dose of 10 or 20 mg/kg was administered intraperitoneally to separate groups of rats 12 days after the administration of IFA. The effects of cepharanthine on RA were determined by measuring serum biochemical parameters, blood cell counts and serum inflammatory and oxidative stress mediators. The expression levels of several proteins that regulate the Wnt/PI3K/Toll-like receptor (TLR)-3 pathway were measured via immunohistochemistry, western blotting and quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) assays. Molecular docking was performed to assess the interactions of cepharanthine with Wnt, PI3K and TLR-3 proteins. Our data revealed that cepharanthine treatment attenuated serum biochemical parameters, blood cell counts and serum inflammatory and oxidative stress mediators in RA rats. Analyses with qRT-PCR and western blotting revealed that the mRNA expression levels of Wnt/PI3K/TLR-3 pathway components were attenuated in cepharanthine-treated RA rats. Cepharanthine also reduced chondrocyte apoptosis and autophagy. Molecular docking analysis revealed significant interactions of cepharanthine with Wnt, PI3K and TLR-3 proteins. In conclusion, the findings implied that cepharanthine treatment attenuates chondrocyte autophagy, apoptosis and proliferation by regulating the Wnt/PI3K/TLR-3 pathway in RA rats.

Phytochemical Screening and Evaluation of the Antiarthritic PotentialofAmmoides pusillaAqueous Extract on Freund's Adjuvant-Induced Rheumatoid Arthritis

Background: Ammoides pusilla plant is a species of therapeutic interest which used in traditional medicines. This work aims to valorize this plant by the characterization of their bioactive components and the evaluation of the in vivo anti-inflammatory potential against a severe disease affecting the bone structure and the stability of the articular cartilage. Methods: First, the phytochemical screening of the polyphenolic extracts of A. pusilla was carried out. The second part of our study is devoted to the evaluation of the in vivo anti-inflammatory activity of the aqueous extract of A. pusilla based on the method of Freund's adjuvant-induced rheumatoid arthritis. Results: Phytochemical tests demonstrated the richness of extract with flavonoids, tannins, coumarins, anthocyanins and triterpenes. Whereas, the quantitative determination reveals that the aqueous extract of A. pusilla is the richest with bioactive components with contents of total polyphenols, flavonoids and tannins equal to 9.52 ± 0.11 mg gallic acid/g, 4.75 ± 0.05 mg quercetin/g and 8.64 ± 0.02 mg catechin/g respectively. The results of the anti-inflammatory activity showed that the aqueous infused extract of A. pusilla has an interesting antiarthritic potential on Freund's adjuvant-induced rheumatoid arthritis in mice. It is manifested by a weight gain; a normal arthritic index and biochemical parameters close to those of Diclofenac®. Conclusion: The aqueous infused extract of A. pusilla is therefore of considerable therapeutic interest as an alternative compound for the prevention of inflammation and for the improvement in bone structure.

Therapeutic effects of celecoxib polymeric systems in rat models of inflammation and adjuvant-induced rheumatoid arthritis.

The incidence of rheumatoid arthritis (RA), an autoimmune inflammatory disease, is rapidly increasing in aging societies. In the current study, celecoxib (CXB) micelles were developed to improve the oral absorption and anti-inflammatory effects of CXB in cell studies and λ-carrageenan rat models, and to enhance the therapeutic effects of CXB on RA in complete Freund's adjuvant (CFA)-induced RA rat models. Moreover, CXB micelles and previously developed solid dispersion (SD6) formulations were evaluated. The physical properties of optimal CXB micelles (M3), such as crystallinity, thermal properties, and intramolecular interactions, were altered. Compared with the commercial product (Celebrex®), the M3 and SD6 formulations showed significantly improved anti-inflammatory effects in terms of nitric oxide reduction, 1.5-fold and 2.2-fold, respectively, at the cellular level. The relative bioavailability (BA) of the M3 and SD6 formulations was also significantly improved as oral bioavailability (167.2% and 219.8% respectively), compared with that of Celebrex®. In particular, M3 and SD6 significantly reduced inflammation and edema volume relative to Celebrex® in CFA-induced RA rat models. Moreover, both M3 and SD6 effectively suppressed CFA-induced pro-inflammatory cytokines (TNF-α and IL-1β) in rat splenic tissues. In conclusion, polymeric systems improved the solubility, relative BA (%) and anti-inflammatory effects of CXB. Thus, CXB polymeric systems show potential as therapeutic agents against inflammation and RA and may need to be tested at the clinical level.

Rhoifolin regulates oxidative stress and proinflammatory cytokine levels in Freund's adjuvant-induced rheumatoid arthritis via inhibition of NF-κB.

Rheumatoid arthritis (RA) is an autoimmune disease of knee joints involving pain and inflammation. Rhoifolin is a plant flavonoid known to have antioxidant and anti-inflammatory properties. This study was taken to identify the effect of rhoifolin on complete Freund’s adjuvant (CFA)-induced arthritis in the rat model. Treatment with rhoifolin (10 and 20 mg/kg) showed a significant improvement in the overall health parameters such as paw edema and weight loss. This improvement in morphological parameters corroborated the findings with gross morphological changes observed in the histopathological analysis. Rhoifolin treatment also caused a significant decrease in oxidative stress, evident from changes in intracellular levels of glutathione, glutathione peroxidase, malondialdehyde, and superoxide dismutase in the articular cartilage tissue. Moreover, proinflammatory cytokines, tumor necrosis factor (TNF)-α, interleukin(IL)-1β, and IL-6 showed a significant downregulation of gene expression and intracellular protein concentration levels. The NF-κB pathway showed a significant attenuation as evident in the significant reduction in the levels of NF-κB p65 and p-IκB-α. These results indicated that rhoifolin can be a natural therapeutic alternative to the extant regimens, which include non-steroidal anti-inflammatory drugs and immunosuppressants. Additionally, the antioxidant and anti-inflammatory action of rhoifolin was probably mediated by the NF-κB pathway. However, the exact target molecules of this pathway need to be determined in further studies.

Anti-arthritic potential of ethanol and aqueous extracts of stem bark of Cleistopholis patens on complete Freund's adjuvant-induced rheumatoid arthritis in rats

BackgroundTraditional medicine intervention has been used in rheumatoid arthritis (RA) treatment due to limitations of conventional drugs. ObjectiveThis study aimed at evaluating the anti-arthritic potentials of ethanol and aqueous extracts of stem bark of Cleistopholis patens (SBCP) in complete Freund's adjuvant (CFA) induced rheumatoid arthritis in rats. Materials and methodsRheumatoid arthritis was induced in groups 2 to 9 by intradermal injection of 0.1 mlkg−1 chicken type II collagen in CFA into the left hind paw of the rats. Group 1 served as normal control. Group 2 (negative control) received 5 mlkg−1 body weight normal saline while group 3 (positive control) received 10 mg/kg body weight standard drug (indomethacin). Groups 4 to 9 received varied doses of the extracts. After 10 days of RA induction, rats were treated with ethanol and aqueous extracts of SBCP orally at a dose of 400, 600 and 800 mgkg−1 for 21 days. The paw size, body weight changes, inflammatory parameters, lipid peroxidation maker and malondialdehyde (MDA) were assessed. ResultsRheumatoid arthritis induction caused marked (p < 0.05) increase in paw size, inflammatory makers and MDA while significant (p < 0.05) reduction was observed in body weight relative to normal control. Treatment with extracts analogous to indomethacin markedly (p < 0.05) decreased the paw size and caused weight gain while the altered inflammatory parameters and MDA were reversed relative to negative control. ConclusionThe findings suggest that the extracts of SBCP have good antiarthritic potentials comparable to indomethacin and hence could be used in rheumatoid arthritis management.