Freund's Adjuvant-induced Rheumatoid Arthritis Research Articles

Exploring the mechanism of Sendeng-4 against rheumacid arthritis through integrated serum pharmacochemistry, transcriptomics, and network pharmacology.

Mongolian medicine Sendeng-4 (SD-4) has demonstrated satisfactory clinical treatment outcomes for rheumatoid arthritis (RA); nevertheless, its bioactive components and the related mechanisms have not yet been clearly elucidated. To explore the bioactive chemical components of SD-4 in the treatment of RA and its possible mechanisms, an High Performance Liquid Chromatography-tandem mass spectrometry (HPLC-MS/MS) method was established to simultaneously quantify the main components in SD-4, and ultraperformance LC-Q-Exactive-MS/MS (UPLC-Q-Exactive-MS/MS) was used to identify the phytochemicals absorbed in the serum. Then, using network pharmacology methods, these components were constructed into a compound-target network of RA to predict possible biological targets of SD-4 as well as potential signaling pathways. Transcriptomics analysis and molecular docking were used to validate the results of network pharmacology. Subsequently, we established a complete Freund's adjuvant-induced RA rat model and observed the anti-RA effects of SD-4 through assessments of foot swelling, ankle diameter, arthritis score, morphology, serum inflammatory factors, and histopathological analysis of synovial tissue. Specifically, reverse transcription-quantitative polymerase chain reaction, Western blot, and immunohistochemical analysis were used in animal experiments to validate the pathways of serum phytochemistry, network pharmacology, and transcriptomics. Tannic acid, gallic acid, corilagin, crocin I, gardenoside, ferulic acid, quercetin, limonin, rutin, chlorogenic acid, verbascoside, catechin, epicatechin, myricetin, and dihydromyricetin in SD-4 showed good linearity within their respective concentration ranges (r ≥ 0.9991); the average recovery rate was 93.77%-109.17% (relative standard deviation < 2%). A total of 37 compounds were identified in serum samples. Based on this, network pharmacology methods collected 739 genes related to these identified compounds in SD-4 and 3807 genes related to RA. Network pharmacology and transcriptomic analysis demonstrated that the phosphatidylinositol 3-kinase (PI3K)-protein kinase B (Akt) signaling pathway is the most relevant pathway affected by SD-4 in RA. In the experiments, SD-4 treatment reduced ankle swelling and arthritis scores in RA rats, improved symptoms, and reduced the production of inflammatory factors. Compared with the RA model group, SD-4 treatment significantly reduced the expression of PI3K-Akt pathway-related messenger RNA and proteins. In addition, immunohistochemical analysis confirmed these results. This study combined serum phytochemistry, network pharmacology, and transcriptomics to demonstrate that SD-4 can alleviate RA by regulating the PI3K-Akt signaling pathway. This research provides a theoretical basis for the clinical application of SD-4 and offers an effective strategy for the identification of bioactive substances in traditional Chinese medicine formulas and the study of their potential mechanisms.

Impact of aqueous leaf-extract of psychotria vogeliana (benth) on serum electrolytes concentration in wistar rats with complete freund's adjuvant-induced rheumatoid arthritis

Plant extracts can influence electrolytes concentration and potentially have implications for rheumatoid arthritis (RA) management. 48 female albino rats weighing between 100 and 200g were used in this study. RA was induced in the rats using 0.1ml of complete Freund’s adjuvant (CFA). Arthritis index was tracked following the Chondrex technique, which only showed signs of inflammation such as redness, edema in the injected paw. Experimental animals were divided into six groups. Group 1 (control) received feeds and water only. Group 2 (toxic) was induced with RA and left untreated. Group 3 (Standard) was induced with RA and treated with standard drug (methotrexate at the dose of 0.225 mg/kg daily). Groups 4, 5 and 6 (treated groups) were induced with RA using CFA, 3 hours later, administered orally with Psychotria vogeliana aqueous leaf-extract at the doses of 200mg/kg, 400mg/kg and 600 mg/kg respectively. Standard methods were employed to determine serum electrolytes concentration. Psychotria vogeliana leaf extract at 200 mg/kg, 400 mg/kg, and 600 mg/kg significantly raised the levels of serum electrolytes of RA rats in a manner comparable to the group that was treated with standard drug and the normal control group. Our findings revealed that rheumatoid arthritis (RA) causes a notable decrease in the concentration of sodium, potassium, calcium, and bicarbonates, which can lead to a depletion of the serum electrolyte balance. Nonetheless, Psychotria vogeliana leaf extract showed a lot of potential for resupplying these vital bodily electrolytes.

In vitro-in vivo assessments of apocynin-hybrid nanoparticle-based gel as an effective nanophytomedicine for treatment of rheumatoid arthritis.

Apocynin (APO), a well-known bioactive plant-based phenolic phytochemical with renowned anti-inflammatory andantioxidant pharmacological activities, has recently emerged as a specific nicotinamide adenine dinucleotide phosphate-oxidase (NADPH) oxidase inhibitor. As far as we know, no information has been issued yet regarding its topical application as a nanostructured-based delivery system. Herein, APO-loaded Compritol® 888 ATO (lipid)/chitosan (polymer) hybrid nanoparticles (APO-loaded CPT/CS hybrid NPs) were successfully developed, characterized, and optimized, adopting a fully randomized design (32) with two independent active parameters (IAPs), namely, CPT amount (XA) and Pluronic® F-68 (PF-68) concentration (XB), at three levels. Further in vitro-ex vivo investigation of the optimized formulation was performed before its incorporation into a gel base matrix to prolong its residence time with consequent therapeutic efficacy enhancement. Subsequently, scrupulous ex vivo-in vivo evaluations of APO-hybrid NPs-based gel (containing the optimized formulation) to scout out its momentous activity as a topical nanostructured system for beneficial remedy of rheumatoid arthritis (RA) were performed. Imperatively, the results support an anticipated effectual therapeutic activity of the APO-hybrid NPs-based gel formulation against Complete Freund's Adjuvant-induced rheumatoid arthritis (CFA-induced RA) in rats. In conclusion, APO-hybrid NPs-based gel could be considered a promising topical nanostructured system to break new ground for phytopharmaceutical medical involvement in inflammatory-dependent ailments.

Research on the improvement effect of Saposhnikovia divaricata (Trucz.) Schischk on rheumatoid arthritis based on the “component-target-pathway” association

ObjectiveTo investigate the therapeutic effect and mechanism of the traditional Chinese medicine Saposhnikovia divaricata (Trucz.) Schischk in rats with complete Freund's adjuvant-induced rheumatoid arthritis (RA). MethodsThe chemical targets and RA targets of Saposhnikovia divaricata (Trucz.) Schischk were acquired by the network pharmacological method. The complete Freund's adjuvant-induced rat RA model was used to further explore the mechanism of Saposhnikovia divaricata (Trucz.) Schischk in improving RA. Pathological changes in the volume of toes, body weight and synovial tissues of joints as well as serum inflammatory factor levels before and after the intervention of Saposhnikovia divaricata (Trucz.) Schischk were investigated. The key metabolic pathways were screened by correlations between metabolites and key targets. Finally, a quantitative analysis of key targets and metabolites was experimentally validated. ResultsSaposhnikovia divaricata (Trucz.) Schischk administration increased body weight, mitigated foot swelling and downregulated inflammatory cytokine levels in model rats. The histopathology showed that treatment with Saposhnikovia divaricata (Trucz.) Schischk can induce inflammatory cell infiltration and synovial hyperplasia and obviously reduce cartilage injuries, thus improving arthritis symptoms in rats. According to the network pharmacology-metabonomics association analysis results, the purine metabolic signaling pathway might be the key pathway for RA intervention with Saposhnikovia divaricata (Trucz.) Schischk. Targeted metabonomics, Western blotting (WB) and reverse transcription-polymerase chain reaction (RT‒PCR) assays showed that the recombinant adenosine deaminase (ADA) mRNA expression level and metabolic level of inosine in Saposhnikovia divaricata (Trucz.) Schischk administration group were lower than those of the model group. This reflected that Saposhnikovia divaricata (Trucz.) Schischk could improve RA by downregulating ADA mRNA expression levels and the metabolic level of inosine in the purine signaling pathway. ConclusionBased on the “component-disease-target” association analysis, this study concludes that Saposhnikovia divaricata (Trucz.) Schischk improves complete Freund's adjuvant-induced RA symptoms in rats mainly by downregulating ADA mRNA expression levels in the purine metabolic signaling pathway, mitigating foot swelling, improving the levels of serum inflammatory factors (IL-1β, IL-6 and TNF-α), and decreasing the ADA protein expression level to intervene in purine metabolism.

“Cerastes snake venom as a promising approach in the management of complete Freund's adjuvant-induced rheumatoid arthritis in rats: Involvement of RANKL and JAK/STAT pathway”

Ethnopharmacological relevanceCerastes is a snake found mainly in the Egyptian desert. Many studies were performed to explain the possible snake venom's pharmacological therapeutic effect in different autoimmune diseases. One of the most common auto-immune diseases is rheumatoid arthritis. Rheumatoid arthritis is characterized by a high release of pro-inflammatory and immune-modulatory cytokines. The reduction of these markers can indicate how effective is the administered drug. Aim of the studyThis study aims to explore the potential pharmacological effects of cerastes venom in experimentally-induced RA in rats using Complete Freund's adjuvant - via different mechanisms - by assessing various tissue and serum parameters. Materials and methodsThe rats were assigned to negative control group, cerastes control group, positive control group, dexamethasone-treated group, infliximab-treated group, and cerastes-treated group. The study ended on the 20th day when serum and tissue samples were prepared for further evaluation of reduced glutathione, malondialdehyde, rheumatoid factor, tumor necrosis factor-α, interleukin-6, and nuclear factor kappa-light-chain-enhancer of activated B cells as well as relative expression of phosphorylated Janus-kinase, phosphorylated signal transducers and activators of transcription, nuclear factor erythroid 2–related factor 2, and receptor activator of nuclear factor Kappa-B ligand. In addition, a histopathological examination of different groups' knees joints, and spleen was done. ResultsThe results showed a significant improvement of arthritis induced in the cerastes-treated group in contrast to the positive control group in all assessed parameters. In addition, significant improvement of arthritis was observed in the histopathological examination of different groups' knees joints, and spleen. ConclusionThese results revealed that cerastes snake venom has potent anti-inflammatory and immunomodulatory effects and can be used in the management of arthritis.

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.

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.

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.

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.

Transdermal delivery of fluvastatin sodium via tailored spanlastic nanovesicles: mitigated Freund's adjuvant-induced rheumatoid arthritis in rats through suppressing p38 MAPK signaling pathway

The current study aimed to encapsulate fluvastatin sodium (FVS), a member of the statins family possessing pleiotropic effects in rheumatoid arthritis (RA), into spanlastic nanovesicles (SNVs) for transdermal delivery. This novel delivery could surmount FVS associated oral encumbrances such as apparent first-pass effect, poor bioavailability and short elimination half-life, hence, accomplishing platform for management of RA. To consummate this objective, FVS-loaded SNVs were elaborated by thin film hydration method, utilizing either Span 60 or Span 80, together with Tween 80 or Brij 35 as an edge activator according to full factorial design (24). Applying Design-Expert® software, the influence of formulation variables on SNVs physicochemical properties and the optimized formulation selection were explored. Additionally, the pharmacokinetic studies were scrutinized in rats. Furthermore, in Freund's adjuvant-induced arthritis, rheumatoid markers, TNF-α, IL-10, p38 MAPK, and antioxidant parameters were measured. The optimum SNVs were nano-scaled spherical vesicles (201.54 ± 9.16 nm), having reasonable entrapment efficiency (71.28 ± 2.05%), appropriate release over 8 h (89.45 ± 3.64%) and adequate permeation characteristics across the skin (402.55 ± 27.48 µg/cm2). The pharmacokinetic study disclosed ameliorated bioavailability of the optimum SNVs gel by 2.79- and 4.59-fold as compared to the oral solution as well as the traditional gel, respectively. Moreover, it elicited a significant suppression of p38 MAPK expression and also significant improvement of all other measured biomarkers. Concisely, the foregoing findings proposed that SNVs can be auspicious for augmenting FVS transdermal delivery for management of RA.

Nicorandil and theophylline can protect experimental rats against complete Freund's adjuvant-induced rheumatoid arthritis through modulation of JAK/STAT/RANKL signaling pathway

Signaling pathways are interesting fields of study of pathogenesis and treatment trials. We elucidated the possible protective effects of nicorandil (15mg/kg/day) and theophylline (20mg/kg/day) on experimentally-induced RA, focusing on the role of JAK (Janus Kinase) / STAT (Signal Transducer and Activator of Transcription) / RANKL (Receptor Activator of Nuclear factor-Kappa B Ligand) / cytokine signaling pathway. Four sets of experiments were performed. First, effect of test agents on normal animals was evaluated. Second, effect of test agents was evaluated on Complete Freund's Adjuvant (CFA; 0.3ml, s.c.)-induced RA to investigate anti-arthritic effect. Third, effect of test agents was evaluated on growth hormone (GH; 2mg/kg/day, s.c.)-induced stimulation of JAK/STAT/RANKL/cytokine signaling pathway to investigate the role of this signaling pathway in their anti-arthritic effect. Fourth, the effect of test agents was performed on CFA/GH-induced RA. To fulfill this purpose, serum anti-citrullinated peptide antibody (ACPA), interleukin-6 (IL-6), and cartilage oligomeric matrix protein (COMP), together with tissue JAK2, STAT3, RANKL, inducible and endothelial nitric oxide synthases (iNOS and eNOS) as well as macrophage inflammatory protein (MIP1α) were estimated using ELISA, Western blotting and PCR techniques, confirmed by a histopathological study. Test agents significantly corrected JAK2, STAT3, RANKL and IL-6 values in animals receiving GH. Additionally, test agents could correct ACPA, IL-6, COMP, JAK2, STAT3, RANKL, iNOS, eNOS and MIP1α levels compared with the respective CFA or CFA/GH controls. These results conclude that nicorandil and theophylline have good anti-arthritic effects related to modulation of JAK/STAT/RANKL signaling pathway. Further clinical trials are claimed.

Granisetron and carvedilol can protect experimental rats againstadjuvant-induced arthritis

Context: Rheumatoid arthritis (RA), a disabling autoimmune disorder of the joints as well as other organs, affects about 1% of population. Unfortunately, all current treatments of RA cause severe gastrointestinal, renal and other complications.Objective: We aimed to evaluate the possible antiarthritic effects of a serotonin 5-HT3 receptor blocker, granisetron, and a nonselective adrenergic receptor blocker, carvedilol, on complete Freund's adjuvant-induced RA in adult female albino rats.Materials and methods: Rats were allocated into a normal control group, an arthritis control group, two reference treatment groups receiving dexamethasone (1.5 mg/kg/day) and methotrexate (1 mg/kg/day), and two treatment groups receiving granisetron (2.5 mg/kg/day) and carvedilol (10 mg/kg/day). Serum-specific rheumatoid, immunological, inflammatory and oxidative stress biomarkers were assessed. A confirmatory histopathological study on joints and spleens was performed.Results: Granisetron administration significantly improved all the measured biomarkers, with the values of rheumatoid factor, matrix metalloproteinase-3, cartilage oligomeric matrix protein, immunoglobulin G, antinuclear antibody and myeloperoxidase being restored back to normal levels. Carvedilol administration significantly improved all biomarkers, with serum MPO value restored back to normal levels.Discussion and conclusions: Serotonin 5-HT3 receptor blockers and adrenergic receptor blockers, represented by granisetron and carvedilol, may represent new promising protective strategies against RA, at least owing to immune-modulator, anti-inflammatory and antioxidant potentials.

The anti-osteoporosis effects of CSN1S2 protein of goat milk and yoghurt on a complete Freund's adjuvant-induced rheumatoid arthritis model in rats

This study aimed to evaluate the anti-osteoporosis effect of CSN1S2 protein from goat milk and yoghurt on a complete Freund's adjuvant (CFA)-induced rheumatoid arthritis (RA) model in rats. Twenty-four rats were randomly divided into six groups: control (untreated) group (C), group treated with CSN1S2 (CM) protein from goat milk, group treated with CSN1S2 protein from goat yoghurt (CY), RA group, RA group treated with CSN1S2 protein from goat milk (RAM), and RA group treated with goat yoghurt (RAY). Mineral elements and mesostructure were analyzed using X-ray fluorescence and scanning electron microscopy. Bone histomorphometry was analyzed using the BoneJ software. One way analysis of variance and Tukey post hoc tests were used to analyze and compare the means of all variables between groups. The phosphorus levels were not significantly different between treatment groups relative to the control group (p > 0.05), but were significantly higher in the CM and RAY groups relative to that observed in the CY group (p < 0.05). CSN1S2 protein of goat milk repaired the collagen structure in the femur trabecular bone. The trabecular thickness and volume were significantly lower in CM and CY groups relative to the control group (p < 0.05). The trabecular volume also decreased significantly in the CM group relative to the control group (p < 0.05). The trabecular thickness was significantly lower in the CY group relative to the CM group (p < 0.05), but the trabecular separation and trabecular volume were significantly greater (p < 0.05). The trabecular volumes were significantly elevated in the RAM and RAY groups relative to the CM group (p < 0.05). The trabecular thickness was significantly higher in the RAY group relative to the CM and CY groups (p < 0.05). CSN1S2 protein of goat milk is better than goat yoghurt in repairing femur crystallization and mesostructure in CFA-induced rheumatoid arthritis in rats.

Protective Effects of Simvastatin and Hesperidin against Complete Freund's Adjuvant-Induced Rheumatoid Arthritis in Rats

Background/Aims: Rheumatoid arthritis (RA) is a disabling autoimmune disease for which most current treatments cause massive complications, thereby limiting treatment dose and duration. The anti-arthritic effects of the 3-hydroxy-3-metylglutary-CoA reductase inhibitor, simvastatin, and the natural flavonoid, hesperidin, were investigated against complete Freund's adjuvant-induced RA in rats. Methods: A normal control group, an arthritis control group, 2 reference treatment groups receiving dexamethasone (1.5 mg/kg/day) and methotrexate (1 mg/kg/day), and 2 treatment groups receiving simvastatin (0.5 mg/kg/day) and hesperidin (200 mg/kg/day) were included in the study. Serum rheumatoid factor, matrix metalloprotinease-3 (MMP-3) and cartilage oligomeric matrix protein (COMP) as specific rheumatoid biomarkers, serum immunoglobulin G (IgG) and antinuclear antibody (ANA) as immunological biomarkers, serum tumor necrosis factor-alpha and interleukin-10 as immunomodulatory cytokines, serum myeloperoxidase (MPO) and C-reactive protein as inflammatory biomarkers, and malondialdehyde (MDA) and glutathione (GSH) as oxidative stress biomarkers were assessed, supported by joint and spleen histopathological study. Results: Simvastatin significantly improved all the measured biomarkers, with MMP-3, COMP, and GSH restored to normal levels. Hesperidin significantly improved all the measured biomarkers, with COMP, IgG, ANA, MPO, MDA and GSH restored to normal levels. Conclusion: Simvastatin and hesperidin may be promising protective agents against RA through immunomodulatory, anti-inflammatory and antioxidant potentials.

Ramipril and haloperidol as promising approaches in managing rheumatoid arthritis in rats

Rheumatoid arthritis (RA) is a challenging autoimmune disorder, whose treatments usually cause severe gastrointestinal, renal and other complications. We aimed to evaluate the beneficial anti-arthritic effects of an angiotensin converting enzyme (ACE) inhibitor, ramipril and a dopamine receptor blocker, haloperidol, on Complete Freund's Adjuvant-induced RA in adult female albino rats. Rats were allocated into a normal control group, an arthritis control group, two reference treatment groups receiving dexamethasone (1.5mg/kg/day) and methotrexate (1mg/kg/day), and two treatment groups receiving ramipril (0.9mg/kg/day) and haloperidol (1mg/kg/day). Serum rheumatoid factor, matrix metalloprotinease-3 (MMP-3) and cartilage oligomeric matrix protein as specific rheumatoid biomarkers, serum immunoglobulin G and antinuclear antibody as immunological biomarkers, serum tumor necrosis factor alpha (TNF-α) and interleukin-10 (IL-10) as immunomodulatory cytokines, serum myeloperoxidase and C-reactive protein as inflammatory biomarkers, as well as malondialdehyde and glutathione reduced (GSH) as oxidative stress biomarkers were assessed. A histopathological study on joints and spleens was performed to support the results of biochemical estimations. Ramipril administration significantly corrected all the measured biomarkers, being restored back to normal levels except for MMP-3, TNF-α and IL-10. Haloperidol administration restored all the measured biomarkers back to normal levels except for TNF-α, IL-10 and GSH. In conclusion, ACE inhibitors represented by ramipril and dopamine receptor blockers represented by haloperidol may represent new promising protective strategies against RA, at least owing to their immunomodulatory, anti-inflammatory and antioxidant potentials.

A new ursane triterpenoic acid and other potential anti-inflammatory and anti-arthritic constituents from EtOAc extracts of Vitellaria paradoxa stem bark

ObjectiveVitellaria paradoxa (shea tree) is used in traditional medicine for the treatment of various ailments, including, inflammation and fever. Therefore the present research investigates the anti-inflammatory and anti-rheumatic effects of V. paradoxa stem bark extracts in rats and the isolation and characterization of its active constituents. MethodsThe anti-inflammatory activity of ethyl acetate extract of V. Paradoxa (VPEE) was evaluated by use of the carrageenan-induced paw oedema model in rats. Moreover, rheumatoid arthritis (RA) was induced by injection of Freund's Completed Adjuvant (FCA) into the subplantar surface of the hind paw of the male Wistar rats. Paw volume was measured plethysmometrically. Joint swelling was measured using electronic vernier caliper. Hot plate test was used to assess the effect of VPEE on hyperalgesia while open field was used to assess the locomotors activity. The relative weight of spleen, liver and thymus was obtained as well as some haematological parameters. Tibiotarsal joint was extracted for histopathology under light microscope. Chemical analysis was carried out by high resolution mass spectrometry and one and two-dimensional NMR techniques. ResultsLC–MS analysis of the EtOAc extract revealed the presence of a new triterpenoid and several known compounds. The structure of the novel compound was elucidated by means of LC–MS and selected 1D and 2D-NMR experiments. The biological effects of ethyl acetate (VPEE), methanol (VPME) and water extracts (VPAE) of V. paradoxa were tested on carrageenan model of acute inflammation and FCA-induced rheumatoid arthritis animal model. In the carrageenan-induced inflammation, VPEE (150mg/kg) significant (66.67%) inhibited the first (after 1h) and the second phase (4–6h) of edema formation. On the Complete Freund's adjuvant-induced rheumatoid arthritis, VPEE at the same dose showed a significantly protective effect. On days 19–28th of treatment, the maximum inflammatory percentage was between 9.60 and 8.91% for the VPEE compared to 30.91–24.29% for the controls. All the extracts significantly reduced the score of arthritis but the maximal reduction was obtained with the VPEE on day 24th of the experimentation. The altered haematological parameters in the arthritic rats were significantly recovered to near normal by the treatment with VPEE at the dose of 150mg/kg. Further histological studies revealed the anti-arthritic activity by preventing cartilage destruction of the arthritic joints of adjuvant arthritic rats. The spleen hypertrophy induced by the FCA was also significantly inhibited. ConclusionThese findings provide pharmacological basis for the application of the VPEE in inflammatory disorders.

Protective effect of CSN1S2 protein of goat milk on ileum microstructure and inflammation in rat-CFA-induced rheumatoid arthritis

ObjectiveTo observe the protective effect of goat milk alpha (S2)-casein (CSN1S2) protein on ileum microstructure and inflammation in rat-complete Freund's adjuvant-induced rheumatoid arthritis model. MethodsTwenty four male Wistar rats were divided into six groups of two models. The body weight, food intake and albumin level of all subjects were calculated. The ileum microstructures were analyzed by scanning electron microscopy. Histopathological analysis was observed by hematoxylin-eosin staining and the level expressions of immunoglobulin E, secretory immunoglobulin A, interleukin-17, interleukin-10, Ki-67 and caspase-9 were measured by using western blotting. ResultsCSN1S2 protein of milk or yogurt could repair the ileum villi of rat arthritis group similar to the normal. The level expressions showed the immunoglobulin E, secretory immunoglobulin A, interleukin-17 and caspase-9 decreased in milk CSN1S2 protein and yogurt CSN1S2 protein rat groups. The level expression of interleukin-10 was increased, and also Ki-67 was significantly increased in milk CSN1S2 protein and yogurt CSN1S2 protein rat groups. CSN1S2 protein of milk and yogurt could increase the body weight and albumin significantly, meanwhile food intake increased but not significantly. ConclusionsCSN1S2 protein of goat milk and yogurt could repair the ileum microstructure, suppress inflammatory process and also increase the body weight, food intake and albumin level. This result indicates that goat CSN1S2 protein may protect the ileum disorder in rheumatoid arthritis disease.

Goat milk CSN1S2 is able to decrease the severity scoring, TNF-a, and RAGE expression in complete Freund's adjuvant-induced rheumatoid arthritis model of rats

This study aimed to elucidate whether CSN1S2 protein of goat's milk was able to inhibit pro-inflammatory cytokine [interleukin (IL)-17 and tumor necrosis factor (TNF)-α], RAGE, and caspase-3 expression in rheumatoid arthritis (RA) rats. A total of 24 Wistar female rats, were randomly assigned into four groups: control group (C), CSN1S2 protein of goat's milk group (CM), CFA-induced RA-rats group (RA), and the RA group treated by CSN1S2 protein of goat's milk (RAM). The severity of erythema and swelling in lower extremities were counted by scoring. IL-17, TNF-α, RAGE, and caspase-3 expression in synovial membranes were analyzed by confocal laser scanning microscopy (CLSM) and western blotting. Erythema and swelling in the RA group was significantly attenuated by goat's milk CSN1S2 (p < 0.05), but did not reach the level in the control group (p < 0.05). The use of CLSM, CSN1S2 protein of goat's milk could decrease the TNF-α, caspase-3, and the number of hyperplasia cells in comparison with the RA group (p < 0.05), to reach the level in the control group (p > 0.05). Western blotting analysis showed that the expression of IL-17, RAGE, TNF-α, and caspase-3 were higher in the RA group compared with the control group. CSN1S2 protein of goat's milk decreased RAGE and TNF-α expression, but increased the IL-17 and caspase-3 expression. In conclusion, CSN1S2 protein of goat's milk decreased erythema, swelling, and inflammation in lower extremities. The CSN1S2 protein of goat's milk also decreased TNF-α and RAGE expression in the synovial membrane of ankle joints. Unfortunately, CSN1S2 protein of goat's milk may induce the production of IL-17. Therefore CSN1S2 protein of goat's milk may provide a nutritional therapy for attenuating the inflammation found in rheumatoid arthritis.

Anti-inflammatory and anti-arthritic activity of a methanol extract from Vitellaria paradoxa stem bark.

Background:Vitellaria paradoxa is a traditional medicinal plant of Cameroon. Several studies on this plant have focused on the cosmetic profile of its fruits. The present study focuses on the anti-inflammatory potency of stem barks extract of this plant.Objective:The objective was to evaluate the effect of methanolic extract of V. paradoxa (VPME) stem barks on inflammatory response in rats.Materials and Methods:Anti-inflammatory effects of VPME were evaluated in acute and chronic (28 days) inflammation induced in Wistar albino rats. The effects on hyperalgesia and locomotors activity were also quantified. The relative weight of lymphoid organs was obtained as well as some hematological parameters.Results:In the carrageenan-induced inflammation, VPME (75 mg/kg) exhibited a significant (66.67%) inhibition after 1 h. On the complete Freund's adjuvant-induced rheumatoid arthritis, VPME showed a significant protective effect with 8.12% inflammation against 25.00% for the control group after 2 days of the treatment. The extract (75 and 150 mg/kg) significantly reduced the score of arthritis with a maximum obtained on day 19th of the experimentation. There was a significant increase in the reaction time of rats on the hot plate as well as the exploratory activities of the animals in the open field. This extract significantly prevented weight, hemoglobin and red blood cells losses, and spleen hypertrophy. A protective action against skin destruction and cartilage erosion was evident. Liquid chromatography-mass spectrometry analysis of the extract revealed the presence of catechins.Conclusions:These findings suggested that V. paradoxa may contribute to the reduction of the inflammatory response.