Complete Freund's Adjuvant-induced Arthritis Research Articles

Potential Anti-Inflammatory Effects of Ethanol Extract of Caryota urens Lour Fruits on Freund's Complete Adjuvant-Induced Rheumatoid Arthritis in Mice

Potential Anti-Inflammatory Effects of Ethanol Extract of Caryota urens Lour Fruits on Freund's Complete Adjuvant-Induced Rheumatoid Arthritis in Mice

Investigation of Thuja orientalis Leaves Extract's Anti-Arthritic Activity in Freund's Complete Adjuvant-Induced Arthritis in Wistar Rats

A severe auto-immune disease that destroys the body's joints and may render a person physically incapable is rheumatoid arthritis. In Ayurveda, herbs have been used to cure a wide range of illnesses. Herbal medicines are widely available and have fewer side effects. As a result, many people find herbal treatments to be intriguing. The Thuja orientalis is a valuable plant for Chinese and Indian medicine. The leaves of Thuja orientalis have long been used in traditional medicine to treat rheumatoid arthritis (RA). However, The current study investigates the anti-arthritic mechanism and phytochemical evaluations of methanolic extract of Thuja orientalis leaves. Researchers employed rats that had Freund's Complete Adjuvant (FCA)-induced arthritis to test the anti-arthritic potential of methanol extracts from Thuja orientalis. Among other things, we have examined paw edoema, body mass index, locomotor activity, haematological observations, biochemical assessment, arthritic score, radiography, etc. Rats were given 0.05ml of FCA subcutaneously to cause arthritis. Four groups of arthritic rats and one group of normal rats, each with six animals, were created. The arthritic rats in Groups IV and V got 200mg per kg and 400mg per kg of thuja orientalis extract, respectively. Diclofenac sodium (10mg per kg) was given as a routine medication to Groups III of the arthritic rats, respectively, and CMC (1ml/kg) was given to Group I of the normal animals. The disease control groups were Group II respectively. Paw volumes were measured using a plethysmometer at 1, 7, 14, 21 days. When compared to arthritis control rats, the Thuja orientalis extract (200 and 400mg/kg orally) significantly (p<0.05, p<0.01) decreased the size of the paws and changed the body weight in CFA rats at day 21. In statistical analysis, all data were reported as mean SEM. The mean values of the test groups and the control group were compared using one-way ANOVA and Dunnett's test. The results of the current investigation showed that a methanol extract from Thuja orientalis leaves may have anti-arthritic properties.

Antiarthritic potential of ethanolic extract of Ixora coccinea leaves on complete Freund's adjuvant-induced arthritis in animal model.

Ixora coccinea leaves possess antioxidant, anti-inflammatory, antinociceptive, antimutagenic, and gastroprotective properties. On this background, its antiarthritic potential was evaluated. The objective is to evaluate the effect of Ethanolic extract of Ixora coccinea leaves (EEICL) on complete Freund's adjuvant-induced arthritis in rats. PG research laboratory, Pharmacology Department, MKCG Medical College, Berhampur, Odisha. Thirty-six Wistar albino rats were randomly distributed into sixgroups (n = 6) as follows: Gr 1 (normal control)-DW p.o, Gr-2 (disease control [DC] - Tween 80 p.o), Gr-3 (piroxicam 0.9 mg/kg p.o), Gr-4 (EEICL-1 g/kg, p.o, Gr 4-EEICL-1.5 g/kg p.o, Gr 5-ED50 (0.82 g/kg) + piroxicam (0.45 mg/kg) p.o. After induction of arthritis, drugs, and vehicles were administered daily from 5th to 25th day. On 0, 5th, 10th, 15th, and 25th day, parameters like body weight, rotarod fall time, paw volume displacement, and arthritis index were measured. On the last day, Erythrocyte sedimentation rate (ESR), tissue malondialdehyde (MDA), and histopathological analysis were done. Analysis of parametric data was done by one-way ANOVA and nonparametric data by Kruskal-Wallis test using graph pad prism 7.0. P < 0.05 was considered statistically significant. EEICL (1.5 mg/kg) showed anti-arthritic effect compared with DC. Rotarod fall-off time 137.5 ± 2.5 sec and body weight (139 ± 12.74 g) were increased significantly. The percentage inhibition of paw volume was increased(52%) whereas arthritic score(0.33), ESR(3.51mm/hr), synovial tissue MDA level (0.62±0.13µmol/gm) and Mankin score(2) were reduced significantly as compared to disease control. EEICL has anti-arthritic potential in rat model.

From co-delivery to synergistic anti-inflammatory effect: Studies on chitosan-stabilized Janus emulsions having chloroquine phosphate and flavopiridol in Complete Freund's Adjuvant induced arthritis rat model

For the first time, the co-delivery of chloroquine phosphate and flavopiridol by intra-articular route was achieved to provide local joint targeting in Complete Freund's Adjuvant-induced arthritis rat model. The presence of paired-bean structure onto the dispersed oil droplets of o/w nanosized emulsions allows efficient entrapment of two drugs (85.86–96.22 %). The dual drug-loaded emulsions displayed a differential in vitro drug release behavior, near normal cell viability in MTT assay, better cell uptake (internalization) and better reducing effect of mean immunofluorescence intensity of inflammatory proteins such as NF-κB and iNOS at in vitro RAW264.7 macrophage cell line. The radiographical study, ELISA test, RT-PCR study and H & E staining also indicated a reduction in joint tissue swelling, IL-6 and TNF-α levels diminution, fold change diminution in the mRNA expressions for NF-κB, IL-1β, IL-6 and PGE2 and maintenance of near normal histology at bone cartilage interface respectively. The results of metabolomic pathway analysis performed by LC-MS/MS method using the rat blood (plasma) collected from disease control and dual drug-loaded emulsions treatment groups revealed a new follow-up study to understand not only the disease progression but also the formulation therapeutic efficacy assessment.

Xuelian injection ameliorates complete Freund's adjuvant-induced acute arthritis in rats via inhibiting TLR4 signaling

BackgroundXuelian injection (XI), a classic preparation extracted from Saussureae Involucratae Herba, has been clinically used to manage rheumatoid arthritis (RA) for nearly twenty years in China. However, the underlying anti-RA mechanism of XI remains unclear. In this study, complete Freund's adjuvant (CFA)-induced acute arthritic model was used to examine the anti-RA effects of XI in vivo. The molecular mechanisms of this action were further investigated using lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. MethodsXI and XI freeze dried powder were characterized by UPLC analysis. CD68 and TLR4 expression in the ankle joints was measured by immunohistochemistry. The secretion of inflammatory mediators was detected by ELISA. The expression levels of TLR4 involved components were measured by Western blotting. The localization of transcription factors was measured by immunofluorescence assay. ResultsXI treatment ameliorated arthritic symptoms induced by CFA in the ankle joints of rats. The serum levels of inflammatory mediators, including TNF-α, MCP-1, and Rantes were decreased by XI treatment. The elevation of CD68 and TLR4 levels in ankle joints caused by CFA was suppressed by XI treatment. Moreover, XI treatment inhibited the secretion of nitric oxide and prostaglandin E2 in LPS-treated RAW264.7 macrophages. The expression of their enzymes iNOS and COX-2 was also decreased after XI treatment. The production of inflammatory mediators, including TNF-α, IL-6, IL-1β, MCP-1, MIP-1α, and Rantes was reduced by XI treatment in LPS-stimulated RAW264.7 cells. The phosphorylation of p38, JNK, ERK, TBK1, IKKα/β, IκB, p65, c-Jun, and IRF3 was reduced after XI treatment. Additionally, the expression levels of nuclear proteins of p65, c-Jun, and IRF3 were inhibited by XI treatment. ConclusionsTaken together, XI possesses potential anti-RA effect and the underlying mechanism may be closely associated with the inhibition of TLR4 signaling. Our findings provide further pharmacological justifications for the clinical use of XI in RA treatment.

Potential therapeutic effect of Salvia coccinea leaf extract in chronic disorders: Myocardial infarction, cataract, and arthritis in rat

Aqueous extract from Salvia coccinea leaf (AESL) has been shown its unique anti-oxidant effect via Nuclear factor (NF)-kappa (κ)B pathway in human monocytic THP-1 ​cells and pharmacological effect on inflammatory diseases like diabetes in rats. Using AESL, which possess antioxidant activity by scavenging radicals and decreasing oxidative stress, would be a promising strategy for the treatment of other inflammatory disorders like myocardial infarction, arthritis, and cataract. This study was designed to evaluate the ameliorating effects of AESL on isoproterenol (Iso)-induced myocardium infarction (MI), selenite-induced cataractogenesis, and complete Freund's adjuvant-induced arthritis in Wistar rats. In this study, AESL ameliorated pathological changes in Iso-induced MI heart muscles and Electro Cardio Gram pattern, 35–80 ​% protection of the cataractogenesis, and prevented the apoptosis induced by selenite in the rat lens. Both pre and post-treatment of AESL (600 ​mg/kg, bw) showed no swelling of the joint and, 35 days treatment showed significant prevention against bony destruction as depicted by less narrowing of joint spaces and soft tissue swelling when compared with control rats. Oral administration of AESL inhibited the Iso-induced increase in serum and myocardial lipid peroxidation, aspartate transaminase, alanine transaminase, creatine kinase, troponin-I, and troponin-T, and lactate dehydrogenase of myocardium infarction rats. Significant increases in the activity of superoxide dismutase, catalase, glutathione peroxidase, and a reduced level of glutathione, were observed in AESL treated cataract rat lens. The results demonstrate that AESL is useful in, in addition to diabetes, controlling myocardium infarction, cataracts, and arthritis.

Anti-arthritic efficacy of Bombax ceiba ethanolic extract in a murine model for rheumatoid arthritis using in vivo, in vitro and radiological analysis.

Rheumatoid arthritis (RA) is a multisystem autoimmune disease that causes discomfort, synovial membrane inflammation, peripheral joint inflammation, morning stiffness, articular tissue loss, and restricted joint movement. In the present study, we aim to explore the anti-arthritic efficacy of Bombax ceiba ethanolic extract in a Freund's Complete Adjuvant-induced arthritis, in murine model. The hot soxhlet method was used to extract dried aerial components of Bombax ceiba using an ethyl alcohol: water (70:30) ratio. Bombax ceiba ethanolic extract at two doses of 200 and 400 mg/kg was investigated in Wistar rats against Freund's full adjuvant-induced chronic immunological arthritis. Anti-arthritis efficacy was studied utilising morphological research (paw volume, paw diameter, and body weight). On the 28th day, the animals were sacrificed, and haematological parameters, pro-inflammatory cytokines (TNF-alpha, IL-6), cell culture, histological and radiological analysis were performed. BCEE inhibited paw oedema significantly (P 0.05) at a dose of 40mg/Kg, which was corroborated by paw volume and diameter, as well as haematological parameters, in Freund's complete adjuvant-induced arthritis model. The BCEE also significantly reduced pro-inflammatory cytokine levels and the histopathological changes caused by Freund's full adjuvant model. BCEE preserves synovial membranes by enhancing health and has shown a significant anti-arthritic activity. Thus, data confirms the traditional usage of Bombax ceiba for arthritis.

Anti-Arthritic and Immunomodulatory Potential of Methanolic, n-Hexane, and Ethyl Acetate Fractions of Bark of Acacia modesta on Complete Freund's Adjuvant-Induced Arthritis in Rats.

Rheumatoid arthritis is an autoimmune disorder and topic of interest for researchers due to its increasing frequency and limited treatment. Acacia modesta Wall is known to treat rheumatic disorders in the traditional system of medicinal plants. Traditional medicines are still required for the treatment of this disease due to the large number of side-effects caused by commercial medicines. In the current study, the antiarthritic potential of methanolic extract (AM-metha), n-hexane (AM-hexa) fraction, and ethyl acetate (AM-etha) fraction of the bark of A. modesta against a complete Freund's adjuvant rat model was evaluated. Evaluation using a digital plethysmometer, macroscopic evaluation, and histopathological evaluation were conducted to determine the paw volume and arthritic scoring. ELISA was performed to assess the PGE2 levels. RT-PCR was used to evaluate the expression levels of MMP2, MMP3, MMP9, NF-κB, IL6, IL1β, TNFα, and VEGF. Biochemical and hematological analyses were also conducted. GC/MS was also carried out to analyze the presence of medicinal compounds. The data revealed a marked reduction in the paw volume, arthritic scoring, and histopathological parameters, indicating the anti-arthritic potential of the plant. Treatment with plant extracts and fractions markedly down-regulated MMP2, MMP3, MMP9, NF-κB, IL6, IL1β, TNFα, and VEGF levels. Similarly, PGE2 levels were also found to be ameliorated in the treatment groups, indicating the immunomodulatory property of plant bark. Plant treatment nearly normalized hematological parameters such as counts of WBCs, RBCs, and platelets, along with Hb content, thereby validating the anti-arthritic activity. GC/MS analysis disclosed the presence of strong anti-inflammatory compounds such as lupeol, oleic acid, and squalene. The study showed that A. modesta possesses anti-arthritic and immunomodulatory potential linked to significant down-regulation of pro-inflammatory and inflammatory biomarkers.

Protective effect of fustin against adjuvant-induced arthritis through the restoration of proinflammatory response and oxidative stress.

Rheumatoid arthritis causes irreparable damage to joints. The present research sought to check fustin's anti-arthritic efficacy against the complete Freund's adjuvant-induced arthritis paradigm in animals by altering the inflammatory response. In the rats, complete Freund's adjuvant was used to trigger arthritis and they received fustin at 50 and 100 mg/kg for 21 days. At regular intervals, the hind paw volume and arthritic score were assessed. After the trial period, hematological, antioxidant, pro-inflammatory cytokines, and other biochemical parameters were estimated. Fustin-treated rats showed the down-regulation of hind paw volume, arthritic score, and altered hematological parameters (TLC, DLC (neutrophil, lymphocyte, monocyte, eosinophil, basophil)). Furthermore, fustin significantly mitigates proinflammatory cytokine (reduced interleukin, tumor necrosis factor-a (TNF-α), IL-6, IL-1β), oxidative stress (attenuated malondialdehyde (MDA), catalase (CAT), glutathione (GSH), superoxide dismutase (SOD)), attenuated production of prostaglandin E2 and myeloperoxidase (MPO) and improved nuclear factor erythroid 2-related factor (Nrf2) action. Fustin led to the benefit in arthritis-prone animals elicited by complete Freund's adjuvant via pro-inflammatory cytokine.

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.

Antiarthritic potential of the butanol fraction of Sesuvium sesuvioides: An in vitro, in vivo, and in silico evaluation.

Sesuvium sesuvioides (Fenzl) Verdc (Aizoaceae) has been traditionally used in the treatment of inflammation, arthritis, and gout. However, its antiarthritic potential has not been evaluated scientifically. The current study was designed to assess the antiarthritic properties of the n-butanol fraction of S. sesuvioides (SsBu) by phytochemical analysis, in vitro and in vivo pharmacological activities, and in silico studies. Phytochemical analysis showed total phenolic contents (90.7 ± 3.02mg GAE/g) and total flavonoid contents (23.7 ± 0.69mg RE/g), and further analysis by GC-MS identified possible bioactive phytocompounds belonging to phenols, flavonoids, steroids, and fatty acids. The in vitro antioxidant potential of SsBu was assessed by DPPH (175.5 ± 7.35mgTE/g), ABTS (391.6 ± 17.1mgTE/g), FRAP (418.2 ± 10.8mgTE/g), CUPRAC (884.8 ± 7.97mgTE/g), phosphomolybdenum (5.7 ± 0.33mmolTE/g), and metal chelating activity (9.04 ± 0.58mg EDTAE/g). Moreover, in the in vitro studies, inhibition (%) of egg albumin and bovine serum albumin denaturation assays showed that the anti-inflammatory effect of SsBu at the dose of 800μg/ml was comparable to that of diclofenac sodium used as a standard drug. The in vivo antiarthritic activity was assessed to determine the curative impact of SsBu against formalin-induced (dose-dependent significant (p < 0.05) effect 72.2% inhibition at 750mg/kg compared to standard; 69.1% inhibition) and complete Freund's adjuvant-induced arthritis (40.8%; inhibition compared to standard, 42.3%). SsBu significantly controlled PGE-2 level compared to the control group (p < 0.001) and restored the hematological parameters in rheumatoid arthritis. Treatment with SsBu significantly reduced oxidative stress by reinstating superoxide dismutase, GSH, and malondialdehyde along with pro-inflammatory markers (IL-6 and TNF-α) in arthritic rats. Molecular docking revealed the antiarthritic role of major identified compounds. Kaempferol-3-rutinoside was found to be more potent for COX-1 (-9.2kcal/mol) and COX-2 inhibition (-9.9kcal/mol) than diclofenac sodium (COX-1, -8.0 and COX-2, -6.5kcal/mol). Out of the 12 docked compounds, two for COX-1 and seven for COX-2 inhibition showed more potent binding than the standard drug. The results from the in vitro, in vivo, and in silico approaches finally concluded that the n-butanol fraction of S. sesuvioides had antioxidant and antiarthritic potential, which may be due to the presence of potential bioactive compounds.

Anti-inflammatory mechanisms of eucalyptol rich Eucalyptus globulus essential oil alone and in combination with flurbiprofen.

Inflammation is the core contributor in the pathogenesis of various acute and chronic illness including appendicitis, bronchitis, arthritis, cancer and neurological diseases. NSAIDs, commonly used medications for inflammatory diseases, on prolonged use cause GI bleeding, ulcers and many more issues. Plant-based therapeutic agents including essential oils in combination with low-dose synthetic drugs have been shown to produce synergistic effects and reduce complications of synthetic drugs. This study was designed to evaluate the anti-inflammatory, analgesic and anti-pyretic properties of Eucalyptus globulus essential oil alone and in combination with flurbiprofen. GC-MS analysis was performed to screen chemical composition of oil. In vitro anti-inflammatory assay (membrane stabilization assay) and in vivo inflammatory acute(carrageenan and histamine-induced paw oedema)and chronic (cotton pellet-induced granuloma and Complete Freund's adjuvant-induced arthritis)modelswere performed to check anti-inflammatoryproperties. Acetic acid-induced algesia and yeast-induced pyrexia models were performed to check analgesic and anti-pyretic properties. qRT-PCR was performed to study the effect of treatments on the expression of inflammatory biomarkers. GC-MS analysis of E. globulus essential oil showed the presence of eucalyptol along with other active biomolecules. 500 + 10mg/kg of oil-drug combination showed significantly (p < 0.05) better in vitro membrane stabilization effects as compared with groups treated with 500mg/kg of E. globulus oil and10mg/kg of Flurbiprofenalone. 500 + 10mg/kg of oil-drug combination showed significantly (p < 0.05) better anti-inflammatory, analgesic and antipyretic effects as compared to 500mg/kg of E. globulus oil alone in all in vivo models. When comparison wasdone between 500 + 10mg/kg of oil-drug combination-treated and 10mg/kg Flurbiprofen-treated group, the former group showed significantly (p < 0.05) better anti-inflammatory and anti-pyretic effects, but there were non-significant differences in the analgesic model. Animal grouptreated with10mg/kg of Flurbiprofen showed significantly (p < 0.05) better anti-inflammatory and analgesic effects than group treated with500mg/kg of oil alone while, there were non-significant differences in anti-pyretic effects. qRT-PCR analysis showed significant (p < 0.05)down-regulation in the expressionof IL-4 and TNF-α in serum samples of animals treated with 500 + 10mg/kg of oil-drug combination as compared to thediseased control (arthritic)group. Overall, the current research demonstrates that Eucalyptus globulus essential oil in combination with flurbiprofen showed better anti-inflammatory, analgesic and anti-pyretic effects than oil and flurbiprofen alone which is attributed to the down-regulation of pro-inflammatory biomarkers (IL-4 and TNF-α). Further studies are required to formulate a stable dosage form and to check the anti-inflammatory efficacy in different inflammatory disorders.

“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.

Evaluation of therapeutic potential of ivermectin against complete Freund's adjuvant-induced arthritis in rats: Involvement of inflammatory mediators.

Rheumatoid arthritis is a chronic systemic inflammatory disease with genetic manifestations. According to recently published case reports, patients taking corticosteroid medication for the management of rheumatoid arthritis develop strongloidiasis and are at high risk of developing associated infections. This study explored the antiarthritic role of ivermectin, a drug used in the treatment of strongyloides and to compare its results with dexamethasone. Thirty-two male Wistar rats were randomly divided into four groups: control, diseased, dexamethasone, and ivermectin groups. Rheumatoid arthritis in all rats except the control group was induced by using complete Freund's adjuvant. After 7days of rheumatoid arthritis induction, animals were treated with dexamethasone 5mg/kg and ivermectin 6mg/kg. Body weight, visual arthritic score, total leukocyte count, differential leukocyte count, proinflammatory genes, and histopathological findings were used to assess the effects of ivermectin on rheumatoid arthritis. Treatment with ivermectin showed a significant reduction in inflammatory cells levels, body weight, and visual arthritic score, indicating an improvement in the degree of inflammation as compared with the diseased group. Treatment with ivermectin and dexamethasone significantly reduced the augmentation in the mRNA expression levels of IL-17, TLR-2, TNF, and NF-κB as a result of arthritic development. Ivermectin treatment also showed a significant reduction in the severity of inflammation and destruction of joints and showed comparable effects to dexamethasone, a corticosteroid used for the treatment of rheumatoid arthritis. Ivermectin has significant antiarthritic properties and can be a novel treatment agent for the management of rheumatoid arthritis patients suffering from strongyloidiasis.

Toddalia asiatica extract attenuates adjuvant-induced arthritis by modulating colon Th17/Treg balance and colony homeostasis

Ethnopharmacological relevanceGiven the adverse effects of the current principal treatments, there is still a great need for effective cures for rheumatoid arthritis (RA), an immune-mediated disease. Toddalia asiatica (L.) Lam is a traditional medicinal herb that can be used for RA treatment because of its anti-inflammatory and analgesic properties. Aim of the studyTo investigate the possible effects of Toddalia asiatica extract (TAE) on intestinal immunity and the intestinal bacterial flora in a rat model of RA. Materials and methodsThe anti-arthritis effect of TAE was evaluated in arthritis rats induced by complete Freund's adjuvant-induced arthritis (AIA). Arthritis index (AI) scores, systemic inflammation scores, histopathologic changes in the colon and ankle were detected by hematoxylin and eosin staining. Western blot analysis was performed to assess the protein expression of IL-17A, RORC, IL-1β, IL-6, FOXP3, IL-10 in the colon. RT-PCR was performed to assess the expression of the colon's mRNA. Finally, changes to the gut microbiome by sequencing 16S rDNA. Microbial function prediction was performed using PICRUSt with the KEGG databases and correlation analysis was carried out by computing Spearman's rank correlations. Resultsdemonstrated that TAE administration at a dose of 3 g/kg dramatically decreased AI scores, systemic inflammation scores, and histopathologic lesions of the ankle and colon in AIA rats. TAE was found to significantly reduce the expression levels of Th17-related proteins and mRNAs (IL-17A, RORC, IL-1β and IL-6) in the colon, while increasing the expression levels of Treg-related proteins and mRNA (IL-10 and FOXP3), which helped restore the balance of Th17/Treg immune cells in the colon. Meanwhile, TAE was also found to be capable of remodeling the gut microbiota in AIA rats. Depleting RA-associated genera and thereby increasing α-diversity enriched the gut microbiota's diversity and shifted the community composition dramatically, leading to the increase of Firmicutes_unclassified, Ruminococcaceae_unclassified, Muribaculum, Subdoligranulum, Lachnospira, Marvinbryantia, and the reduction of RA-related bacteria Ligilactobacillus, Streptococcus and Eubacterium-eligens-group. Furthermore, PICRUSt analysis revealed that metabolic pathways were associated with TAE treatment, with metabolic pathways dominating. Among them, metabolic pathways were predominant. Correlation studies showed that a total of 9 microorganisms, including Ligilactobacillus, Eubacterium-eligens-group and Subdoligranulum, were significantly associated with Th17/Treg expression. ConclusionsThis study demonstrates that TAE is a low-toxicity poly alkaline drug that can rapidly and effectively improve joint symptoms in RA rats and increases beneficial intestinal bacteria and decreases harmful ones, which is associated with modulating Th17/Treg interactions in intestinal T cells and reversing microbial disorders.

A Novel Hydroxyapatite/Vitamin B12 Nanoformula for Treatment of Bone Damage: Preparation, Characterization, and Anti-Arthritic, Anti-Inflammatory, and Antioxidant Activities in Chemically Induced Arthritic Rats.

The usage of nanomaterials for rheumatoid arthritis (RA) treatment can improve bioavailability and enable selective targeting. The current study prepares and evaluates the in vivo biological effects of a novel hydroxyapatite/vitamin B12 nanoformula in Complete Freund's adjuvant-induced arthritis in rats. The synthesized nanoformula was characterized using XRD, FTIR, BET analysis, HERTEM, SEM, particle size, and zeta potential. We synthesized pure HAP NPs with 71.01% loading weight percentages of Vit B12 and 49 mg/g loading capacity. Loading of vitamin B12 on hydroxyapatite was modeled by Monte Carlo simulation. Anti-arthritic, anti-inflammatory, and antioxidant effects of the prepared nanoformula were assessed. Treated arthritic rats showed lower levels of RF and CRP, IL-1β, TNF-α, IL-17, and ADAMTS-5, but higher IL-4 and TIMP-3 levels. In addition, the prepared nanoformula increased GSH content and GST antioxidant activity while decreasing LPO levels. Furthermore, it reduced the expression of TGF-β mRNA. Histopathological examinations revealed an improvement in joint injuries through the reduction of inflammatory cell infiltration, cartilage deterioration, and bone damage caused by Complete Freund's adjuvant. These findings indicate that the anti-arthritic, antioxidant, and anti-inflammatory properties of the prepared nanoformula could be useful for the development of new anti-arthritic treatments.

Euphorbium total triterpenes improve Freund's complete adjuvant-induced arthritis through PI3K/AKT/Bax and NF-κB/NLRP3 signaling pathways.

Euphorbium is the resinous secretion of Euphorbia resinifera Berg. According to the record, Euphorbium was first used by Roman doctors to treat the emperor's joint pain. In China, it is applied in folk medicine to treat damp-cold or mucous diseases, such as arthralgia and ascites, etc. This herb is used for rheumatoid arthritis and skin tumors in the folklore of northeastern Brazil. Triterpenes are mainly characteristic constituents of Euphorbium, and possibly possess anti-rheumatoid arthritis. To explore the preventive effect of Euphorbium total triterpenes (TTE) on Freund's complete adjuvant (FCA) induced arthritis in rats and its mechanism. TTE was extracted and isolated from Euphorbium, and its components were analyzed by HPLC. The safety of TTE was evaluated by an acute toxicity test in mice. Arthritis was induced in rats by injecting 0.2mL FCA into the right hind paw toe, except for the control group, which was given the same volume of physiological saline. Tripterygium Glycosides (TG, 7.5mg/kg) and TTE (32, 64 and 128mg/kg) were administered by gavage for 30 days. Body weights, paw swelling, and arthritic scores were measured during the experiment process. After 30 days, blood and joints were harvested to determine various indicators of arthritis. The contents of euphol and euphorbol in TTE were 47.03% and 18.77% respectively, and the maximal feasible dose of TTE in mice is 12 g/kg. The experimental results showed that arthritis indicators in rats deteriorated after FCA inducement compared with the control group. After treatment with TTE, the swelling degree and histopathological change of the hind paws in rats were significantly improved as well as arthritic score; the serum TNF-α, CRP, IL-1β, IL-6, IL-18 and RF levels in rats were significantly reduced; The expression of PI3K, AKT, P-AKT, Bcl-2, NF-κB, NLRP3 and Pro-caspase-1 protein in joint tissue were down-regulated, and the expression of Bax protein was up-regulated. The results suggested that TTE possessed anti-arthritis effects, and its mechanism may be related to its anti-inflammatory and immunomodulatory properties, as well as regulation of PI3K/AKT/Bax and NF-κB/NLRP3 signaling pathway.

Anti-inflammatory effects of Solanum procumbens on a low dose complete Freund's adjuvant-induced arthritis rat model

Objective: To investigate the anti-inflammatory and analgesic effects of Solanum procumbens on complete Freund's adjuvant-induced arthritis rat models.Methods: We isolated and identified five compounds in the ethanol-soluble Solanum procumbens extract (SP) with anti-inflammatory effects, including ursolic acid, β-sitosterol, hexadecanoic acid, cis-vaccenic acid, and vanillic acid. Additionally, we investigated the anti-inflammatory effects of SP on rheumatoid arthritis symptoms, including paw volumes, local temperatures, withdrawal latency, and mechanical withdrawal threshold at the hind paw and white blood cell (WBC) number from complete Freund's adjuvant-induced arthritis rat models.Results: We have successfully established a complete Freund's adjuvant-induced arthritis rat model at a low dose (1 mg/mL). SP extract significantly reduced paw volumes (P<0.05), prolonged withdrawal latencies (P<0.05), decreased local temperature, and increased the mechanical withdrawal threshold (P<0.05), but only SP extract at the dose of 300 mg/kg significantly decreased WBC numbers.Conclusions: SP extract could be a potential medication candidate with anti-inflammatory effects for arthritis, but it requires further investigation into the mechanism of the SP and its effectiveness on other models as well as clinical trials.

Evaluation of the anti-arthritic activity of Rhuflex-F - A proprietary Ayurvedic herbomineral formulation in albino rats.

Rhuflex-F is a proprietary Ayurvedic herbo-mineral formulation clinically used to combat and relieve stiffness in joints and muscles, reduce edema, restore mobility, and also effective in relieving the symptoms of other autoimmune illnesses that lead to rheumatism. The aim and objective of the research study is to evaluate the efficacy of Rhuflex-F against in vitro protein denaturation and in vivo Freund's adjuvant-induced arthritis in albino rats. In vitro inhibition of protein denaturation activity was carried out using bovine serum albumin. For in vivo activity, arthritis was induced by complete Freund's adjuvant in albino rats. Rhuflex-F (135-270 mg/kg, po) was administered for 30th days in arthritic rats, and effects were assessed on primary and secondary paw edema, on pain response, hematological, serum biochemical parameters (serum transaminases, alkaline phosphatase, urea, uric acid, and orosomucoid), and serum anti-oxidant parameters and adrenal ascorbic acid. Aqueous extract of Rhuflex-F showed in vitro protein denaturation inhibitory activity in a dose-dependent manner. Rhuflex-F showed nonsignificant decrease in primary and secondary paw edema with reduced pain response, some reversal effects on hematological parameters such as white blood cell and red blood cell related parameters and serum orosomucoid and adrenal ascorbic acid in comparison to Fruend's adjuvant control group. Further, Rhuflex-F reversed Freund's adjuvant-induced adverse effects on oxidant status in the serum of albino rats. Result of the present study suggested that Rhuflex-F formulation has anti-inflammatory activity, may be due to the inhibition of protein denaturation in vitro and in vivo anti-arthritic activity against complete Freund's adjuvant-induced arthritis in albino rats.