Rat Paw Volume Research Articles

Investigating Skin Permeation Behavior, Skin Irritation and Anti-Inflammatory Activity of Diclofenac Diethylamine Novel Formulation Prepared using Shea Butter as Absorption Base with Nerolidol as Permeability Enhancer

Background: The present investigation was to study the drug permeation of diclofenac diethylamine (DDEA) cream prepared using shea butter as the absorption base and nerolidol as a permeability enhancer and also to evaluate skin irritation and anti-inflammatory study of prepared cream on Wistar rats in comparison with marketed formulation. Materials and Methods: The cream prepared using shea butter and nerolidol was assessed for permeation study through rat skin. The three formulations mainly F0 (without nerolidol), F5 (containing 0.5% nerolidol), and the marketed formulation were assessed for % drug permeation through rat skin along with an estimation of flux and permeability coefficient. A skin irritation study of a placebo, F0, and F5 formulation was carried out on Wistar rats. Anti-inflammatory study of transdermal cream formulation of control, placebo, F0, F5, and marketed formulation was evaluated for anti-inflammatory study on Wistar rat using carrageenan-induced rat paw edema method. Results: The F5 formulation showed enhanced drug permeation as compared to the F0 and marketed formulation. The flux value F5 formulation was found to be 0.3942 ± 0.009 g/cm2/min as compared to F0 and marketed formulation which were found to be 0.2789 ± 0.013 g/cm2/min and 0.2730 ± 0.0110 g m/cm2/min, respectively. The permeability coefficient for the F5 formulation was found to be 2.370 × 10-5 m/s as compared to F0 and marketed formulations which were found to be 1.680 × 10-5 and 1.640 × 10-5 m/s, respectively. The F0 and F5 formulations were found to be non-irritant and no edema was observed on its application. F5 formulation showed significant inhibition of edema in rat paw volume induced by carrageenan as compared to control, placebo, Std, and F0 batches. Conclusion: The study demonstrates that the F5 formulation prepared using shea butter as an absorption base and containing 0.5% w/v of nerolidol as a permeability enhancer showed a better permeation of DDEA through rat skin, non-irritant in nature and significant anti-inflammatory activity as compared to F0 and marketed formulation.

Inhibition of alternative complement system and prolyl hydroxylase ameliorates anaemia of inflammation.

Chronic diseases associated with inflammation cause early destruction of RBCs. Complement system, part of innate immunity, is involved in such RBC destruction. Persistent inflammation causes kidney injury, leading to reduced erythropoietin release and functional iron deficiency, causing anaemia. We have investigated effect of iptacopan, a factor B inhibitor, and desidustat, a prolyl hydroxylase inhibitor in anaemia induced by peptidoglycan polysaccharide (PGPS) treatment in rats. Inflammation, haemolysis and its diagnostic markers (LDH, total bilirubin, and RBC lifespan) were evaluated after three days of PGPS challenge. Haemoglobin, RBC, iron homeostasis, and RBC destruction were evaluated fourteen days after PGPS challenge. Desidustat (15mg/kg) and iptacopan (20mg/kg) were given along with PGPS and continued for two weeks. Iptacopan and its combination with desidustat prevented LDH, total bilirubin, complement protein-C3a and haemolysis. Combination treatment caused an early normalization of haemoglobin and RBC. Combination also reduced WBC, alkaline phosphatase, aspartate aminotransferase, and rat paw volume. Serum iron was increased by desidustat and its combination treatment. Spleen weight, tissue iron, and serum hepcidin were reduced by combination treatment. Effect of desidustat alone was prominent on iron (serum and tissue) and hepcidin. Thus, combination of iptacopan and desidustat can be a potentially useful therapeutic option for treatment of anaemia of inflammation.

Artemisia pallens W. Attenuates Inflammation and Oxidative Stress in Freund's Complete Adjuvant-Induced Rheumatoid Arthritis in Wistar Rats.

Rheumatoid arthritis (RA) is an autoimmune disease that causes distinctive inflammatory symptoms and affects over 21 million people worldwide. RA is characterized by severe discomfort, swelling, and degradation of the bone and cartilage, further impairing joint function. The current study investigates the antiarthritic effect of a methanolic extract of Artemisia pallens (methanolic extract of A. pallens, MEAP), an aromatic herb. Artemisinin content (% per dry weight of the plant) was estimated using a UV Vis spectrophotometer. In the present study, animals were divided into six groups (n = 6). The control group (group I) was injected with 0.25% of carboxymethyl cellulose. The arthritic control group (group II) was treated with Freund's complete adjuvant (by injecting 0.1 mL). Prednisolone (10 mg/kg), a lower dose of MEAP (100 mg/kg), a medium dose of MEAP (200 mg/kg), and a higher dose of MEAP (400 mg/kg) were orally delivered to groups III, IV, V, and VI, respectively. Freund's complete adjuvant was administered into the sub-plantar portion of the left-hind paw in all the groups except vehicle control to induce rheumatoid arthritis. Weight variation; joint diameter; paw volume; thermal and mechanical hyperalgesia; hematological, biochemical, and oxidative stress parameters; radiology; and a histopathological assessment of the synovial joint were observed in order to evaluate the antiarthritic effect of the methanolic extract of A. pallens. In this study, the estimated content of artemisinin was found to be 0.28% (per dry weight of the plant), which was in good agreement with the reported value. MEAP (200 and 400 mg/kg) caused a significant reduction in increased paw volume and joint diameter in arthritic rats while significantly increasing body weight and the mechanical threshold of thermal algesia. Moreover, complete blood counts and serum enzyme levels improved significantly. Radiological analysis showed a reduction in soft tissue swelling and small erosions. A histopathological examination of the cells revealed reduced cell infiltration and the erosion of joint cartilage in MEAP-administered arthritic rats. The present research suggests that the antiarthritic activity of the methanolic extract of A. pallens wall is promising, as evidenced by the findings explored in our rat model.

Cayratia albifolia C.L.Li exerts anti-rheumatoid arthritis effect by inhibiting macrophage activation and neutrophil extracellular traps (NETs)

BackgroundCayratia albifolia C.L.Li (CAC), commonly known as “Jiao-Mei-Gu” in China, has been extensively utilized by the Dong minority for several millennia to effectively alleviate symptoms associated with autoimmune diseases. CAC extract is believed to possess significant anti-inflammatory properties within the context of Dong medicine. However, an in-depth understanding of the specific pharmaceutical effects and underlying mechanisms through which CAC extract acts against rheumatoid arthritis (RA) has yet to be established.MethodsTwenty-four Sprague–Dawley rats were divided into four groups, with six rats in each group. To induce the collagen-induced arthritis (CIA) model, the rats underwent a process of double immunization with collagen and adjuvant. CAC extract (100 mg/kg) was orally administered to rats. The anti-RA effects were evaluated in CIA rats by arthritis score, hind paw volume and histopathology analysis. Pull-down assay was conducted to identify the potential targets of CAC extract from RAW264.7 macrophage lysates. Moreover, mechanism studies of CAC extract were performed by immunofluorescence assays, real-time PCR and Western blot.ResultsCAC extract was found to obviously down-regulate hind paw volume of CIA rats, with diminished inflammation response and damage. 177 targets were identified from CAC extract by MS-based pull-down assay. Bioinformatics analysis found that these targets were mainly enriched in macrophage activation and neutrophils extracellular traps (NETs). Additionally, we reported that CAC extract owned significant anti-inflammatory activity by regulating PI3K-Akt-mTOR signal pathway, and inhibited NETosis in response to PMA.ConclusionsWe clarified that CAC extract significantly attenuated RA by inactivating macrophage and reducing NETosis via a multi-targets regulation.Graphical

Effect of different suspension moxibustion methods on syndrome characteristics of rats with rheumatoid arthritis of heat bi syndrome based on "moxibustion can be used for heat syndrome"

To observe the effects of different suspension moxibustion methods on the syndrome characteristics and inflammatory factors of rats with rheumatoid arthritis (RA) of heat bi syndrome and to prove the concept of "moxibustion can be used for heat syndrome". Among seventy Wistar rats, 12 rats were randomly selected as a normal group, and the remaining rats were induced by collagen combined with wind, dampness, and heat environmental stimulation to establish the RA model of heat bi syndrome. Forty-eight rats with successful model establishment were further randomly divided into a model group and three moxibustion groups (mild moxibustion group, rotating moxibustion group and sparrow-pecking moxibustion group), with 12 rats in each group. The acupoints "Quchi" (LI 11), "Dazhui" (GV 14) and ashi point were used in all moxibustion groups, with mild moxibustion, rotating moxibustion, and sparrow-pecking moxibustion intervention given respectively, each acupoint was treated with moxibustion for 10 min a day, and 6 days were considered one course of treatment, with a total of three courses. After the intervention, the arthritis index (AI), the Evans blue (EB) extravasated volume in the soft tissue of the right hind paw, and the levels of tumor necrosis factor (TNF)-α and interleukin (IL)-10 in the serum were measured by ELISA in each group. The volume of the bilateral hind paw was measured; the infrared thermal imaging was collected to analyze the temperature of the plantar area of the bilateral foot pads, and the reaction time of plantar heat pain was calculated before and after modeling, as well as after the 1st, 2nd and 3rd courses of interrention. The ankle dorsiflexion angle of the right hind foot was also measured before and after modeling, as well as after the intervention. After modeling, compared with the normal group, the rats in the model group had more high-temperature areas in the bilateral hind limbs, abnormal AI score, abnormal bilateral hind paw volume, abnormal temperature of the plantar area of the bilateral foot pads, abnormal foot pain response time, abnormal right hind ankle dorsiflexion angle, abnormal right hind paw soft tissue EB extravasation, and abnormal serum TNF-α and IL-10 levels (P<0.01, P<0.05). After the intervention, compared with the model group, the rats in each moxibustion group had decreased or disappeared high-temperature areas in the bilateral hind limbs, EB extravasated volume in the soft tissue of the right hind paw was reduced (P<0.05), and the right ankle dorsiflexion angle was increased (P<0.05), serum level of TNF-α was reduced, and level of IL-10 increased (P<0.05); the AI scores in the mild moxibustion group and the sparrow-pecking moxibustion group was decreased (P<0.01, P<0.05). After the 1st, 2nd and 3rd courses of intervention, compared with the model group, the bilateral hind paw volume of rats in each moxibustion group was decreased (P<0.05, P<0.01), and plantar heat pain reaction time was increased (P<0.05). After the 2nd course and the 3rd course of intervention, the temperature of the right hind paw pad area was decreased in each moribustion group (P<0.05); after the 3rd courses of intervention, the temperature of the left hind paw pad area was decreased in the mild moxibustion group (P<0.05). Suspension moxibustion could adjust the serum levels of TNF-α and IL-10 to improve the syndrome characteristics of RA rats of heat bi syndrome, such as joint redness, swelling, heat, pain and activity restriction. The effect of mild moxibustion is the most prominent. The findings could provide scientific basis for "moxibustion can be used for heat syndrome".

The immunosuppressive effects and mechanisms of loureirin B on collagen-induced arthritis in rats.

Rheumatoid arthritis (RA) is a common disease mainly affecting joints of the hands and wrists. The discovery of autoantibodies in the serum of patients revealed that RA belonged to the autoimmune diseases and laid a theoretical basis for its immunosuppressive therapy. The pathogenesis of autoimmune diseases mainly involves abnormal activation and proliferation of effector memory T cells, which is closely related to the elevated expression of Kv1.3, a voltage-gated potassium (Kv) channel on the effector memory T cell membrane. Drugs blocking the Kv1.3 channel showed a strong protective effect in RA model animals, suggesting that Kv1.3 is a target for the discovery of specific RA immunosuppressive drugs. In the present study, we synthesized LrB and studied the effects of LrB on collagen- induced arthritis (CIA) in rats. The clinical score, paw volume and joint morphology of CIA model rats were compared. The percentage of CD3+, CD4+ and CD8+ T cells in rat peripheral blood mononuclear and spleen were analyzed with flow cytometry. The concentrations of inflammatory cytokines interleukin (IL)-1b, IL-2, IL-4, IL-6, IL-10 and IL-17 in the serum of CIA rats were analyzed with enzyme-linked immunosorbent assay. The IL-1b and IL-6 expression in joints and the Kv1.3 expression in peripheral blood mononuclear cells (PBMCs) were quantified by qPCR. To further study the mechanisms of immunosuppressive effects of LrB, western blot and immunofluorescence were utilized to study the expression of Kv1.3 and Nuclear Factor of Activated T Cells 1 (NFAT1) in two cell models - Jurkat T cell line and extracted PBMCs. LrB effectively reduced the clinical score and relieved joint swelling. LrB could also decrease the percentage of CD4+ T cells, while increase the percentage of CD8+ T cells in peripheral blood mononuclear and spleen of rats with CIA. The concentrations of inflammatory cytokines interleukin (IL)-1b, IL-2, IL-6, IL-10 and IL-17 in the serum of CIA rats were significantly reduced by LrB. The results of qPCR showed that Kv1.3 mRNA in the PBMCs of CIA rats was significantly higher than that of the control and significantly decreased in the LrB treatment groups. In addition, we confirmed in cell models that LrB significantly decreased Kv1.3 protein on the cell membrane and inhibited the activation of Nuclear Factor of Activated T Cells 1 (NFAT1) with immune stimulus. In summary, this study revealed that LrB could block NFAT1 activation and reduce Kv1.3 expression in activated T cells, thus inhibiting the proliferation of lymphocytes and the release of inflammatory cytokines, thereby effectively weakening the autoimmune responses in CIA rats. The effects of immunosuppression due to LrB revealed its potential medicinal value in the treatment of RA.

Combined treatment with glucosamine and chondroitin sulfate improves rheumatoid arthritis in rats by regulating the gut microbiota

BackgroundTo investigate the ameliorative effects of glucosamine (GS), chondroitin sulphate (CS) and glucosamine plus chondroitin sulphate (GC) on rheumatoid arthritis (RA) in rats, and to explore the mechanism of GS, CS and GC in improving RA based on the gut microbiota.MethodsRA rat models were effectively developed 14 days after CFA injection, and then garaged with GS, CS and GC. Body weight and paw volume of rats were monitored at multiple time points at the beginning of CFA injection. Until D36, serum and ankle tissue specimens were used to measure levels of circulating inflammatory factors (TNF-α, IL-1β, MMP-3, NO and PGE2) and local inflammatory indicators (TLR-4 and NF-κB). On D18, D25, and D36, intergroup gut microbiota was compared using 16S rRNA gene sequencing and bioinformatics analysis. We also performed the correlation analysis of gut bacteria, joint swelling and inflammatory indicators.ResultsGC, rather than GS and CS, could reduce right paw volumes, levels of TLR-4 and NF-κB in synovial tissues. In addition, enriched genera in RA model rats screened out by LEfSe analysis could be inhibited by GC intervention, including potential LPS-producing bacteria (Enterobacter, Bacteroides, Erysipelotrichaceae_unclassified and Erysipelotrichaceae_uncultured) and some other opportunistic pathogens (Esherichia_Shigella, Nosocomiicoccus, NK4A214_group, Odoribacter, Corynebacterium and Candidatus_Saccharimonas.etc.) that positively correlated with pro-inflammatory cytokines, right paw volume, and pathology scores. Furthermore, the gut microbiota dysbiosis was observed to recover before alleviating joint swelling after interventions.ConclusionsGC could inhibit potential LPS-producing bacteria and the activation of TLR-4/NF-κB pathway in RA rats, thus alleviating RA-induced joint injury.

Immunomodulatory Effects of Phaleria macrocarpa Leaf Extract in Normal And Cyclophosphamides Induced in Wistar Rats

Abstract. Cyclophosphamide is an antineoplastic drug belonging to the alkylating agents commonly used in treating cancer. However, the use of cyclophosphamide causes a decrease in the body's immune system by reducing lymphocyte proliferation. The curent study was conducted to evaluate the immunomodulatory effect of ethyl acetate extracts of the leaf of the P. macrocarpa (EEADMD) and the ethanol extracts of the leaf of the P. macrocarpa (EEDMD) in normal and cyclophosphamide induced rats. This study used rats classified into two major groups: normal rats and rats induced by cyclophosphamide which were then given P. macrocarpa leaf extract until day 14. On day 4, the test animals were infected with 1% Staphylococcus aureus suspension in. Induced with cyclophosphamide 70 mg/kg BW was carried out on the 8th and 13th days, and then the immunomodulatory activity was tested using total leukocyte analysis, leukocyte differential, and delayed type hypersensitivity response. The results showed that EEDMD 400 mg/kg BW in normal rats and EEDMD 100 mg/kg BW in cyclophosphamide induced rats could increase total leukocytes and leukocyte differential with a significant difference to the negative group (p &lt;0.05). The results of the delayed hypersensitivity response test of EEADMD and EEDMD at a dose of 100 mg/kg BW with cyclophosphamide induction and normal rats could give an increase in rat paw volume with a significant difference to the negative group (p &lt;0.05). This study shows that EEADMD and EEDMD have an immunomodulatory effect on increasing total and differential leukocytes and leukocyte differential and delayed type hypersensitivity response.

Effect of Trichilia monadelpha (Thonn.) J. J. de Wilde (Meliaceae) extracts on C-reactive proteins levels and acute inflammation

Trichilia monadelpha is reported to have acute anti-inflammatory effects. This study sought to establish the possible effects of the petroleum ether, ethyl acetate, and ethanolic extracts of Trichilia monadelpha stem bark (i.e., PEE, EAE, and EthE, respectively) on C-reactive proteins, using various acute inflammatory models. Changes in hind paw volume of Sprague-Dawley rats were measured before and after intraplantar injection of; 50 μL of 0.5% λ-carrageenan, 0.1 ml of 0.1% histamine, 0.1 ml of 0.02% serotonin, 1 nmol Prostaglandin E2, or 10 nmol bradykinin, to induce paw oedema, and during treatment with 10, 30, or 100 mgkg−1 PEE, EAE, or EthE, or various reference drugs. Changes in rectal temperature, and plasma C-reactive protein levels were also measured after injecting, intraperitoneally, 0.1 mgkg−1 Lipopolysaccharide (Escherichia coli) to induce pyrexia.PEE, EAE, and EthE reduced significantly (P ≤ 0.01–0.001; maximum inhibition at 100 mgkg−1 (imax) = 55.0–76.7%) paw oedema induced by carrageenan; comparable to diclofenac (P ≤ 0.0001; imax = 93.1%). PEE, EAE and chlorpheniramine prevented significantly (P ≤ 0.0001; imax = 115–144%) or reduced significantly (P ≤ 0.0001; imax = 80–85%) histamine-induced paw oedema. PEE, EAE and ondansetron significantly decreased (curative: P ≤ 0.01–0.001, imax = 51–82%; prophylactic: P ≤ 0.01–0.001, imax = 56–77%) serotonin-induced inflammation. In prostaglandin E2 –induced inflammation PEE and EAE significantly reduced (curative: P ≤ 0.05–0.01, imax = 60–70%; prophylactic: P ≤ 0.0001, imax = 86–332%) oedema; PEE and EAE also significantly decreased (curative: P ≤ 0.050–0001, imax = 46–98%; prophylactic: P ≤ 0.01–0.001 imax = 62–78%) in bradykinin-induced inflammation. PEE, EAE, Diclofenac and Dexamethasone significantly reduced (P ≤ 0.05–0.001) plasma C-reactive protein levels. PEE, EthE, EAE and Aspirin significantly reduced (P ≤ 0.05–0.001) febrile response.Trichilia monadelpha stem bark extracts had anti-inflammatory and anti-pyretic properties and significantly reduced C-reactive proteins associated with carrageenan-induced inflammation.

Amelioration of complete Freund\u2019s adjuvant-induced arthritis by Calotropis procera latex in rats

BackgroundRheumatoid arthritis is the most common cause of disability, affecting 0.3–1% of the adult population worldwide. The latex of Calotropis procera possesses potent anti-inflammatory as well as analgesic properties. In light above facts, the present study was designed to evaluate anti-arthritic activity of Calotropis procera latex in complete Freund's adjuvant (CFA)-induced arthritis in Wistar albino rats. Complete Freund's adjuvant was injected into the left hind paw on day 0, and treatment of prednisolone and Calotropis procera latex was given from day 0 to 28. Various biochemical, hematological and functional parameters as well as radiological and histopathological changes of joint along with body weight and paw volume were measured.ResultsCalotropis procera treatment significantly lowered paw volume in CFA-induced arthritic rats. Significant improvement was observed in functional, biochemical and hematological parameters in Calotropis procera-treated rats. However, the body weight remained unaffected. Histological and radiographical examination of synovial joints in Calotropis procera-treated animals exhibited less synovial hyperplasia, infiltration and accumulation of inflammatory cell in synovial fluid, cartilage and bone erosion and joint space narrowing.ConclusionCalotropis procera latex possesses anti-arthritic activity, which is facilitated by modulation in the level of inflammatory mediators and oxidative stress. The improvement in hematological as well as biochemical parameters might be reflected on functional, histopathological, radiological changes and thereby disease progression.

Modulation of psoriatic-like skin inflammation by traditional Indian medicine Divya-Kayakalp-Vati and Oil through attenuation of pro-inflammatory cytokines

Background and aimPsoriasis (Ps) is a chronic skin inflammatory disorder, that progresses to scaly-red dermal plaque formations associated with inflammation. Divya-Kayakalp-Vati (DKV) and Divya-Kayakalp-Oil (DKO) are traditional Ayurveda herbo-mineral formulations, that are prescribed for the treatment of inflammatory dermal ailments. In the present study, we evaluated the efficacy of Divya-Kayakalp-Vati and Divya-Kayakalp-Oil (DKV-O) combined treatment in ameliorating Ps-like skin inflammation under in-vitro and in-vivo conditions. Experimental procedureEfficacy of DKV-O was analyzed in λ-carrageenan-treated Wistar rats paw edema and 12-O-tetradecanoylphorbol 13-acetate (TPA)-treated CD-1 mouse ear edema models through physiological and histopathological analysis. Mode of action for the DKV-O was studied in LPS-stimulated THP-1 cells through pro-inflammatory cytokine analysis. Observed effects were correlated to the phytochemicals constituents of DKV-O analyzed using the HPLC method. Result and conclusionDKV and DKO formulations were individually found to contain phytochemicals Gallic acid, Catechin, Berberine, Curcumin, Phenol and Benzoic acid. DKV-O treatment significantly reduced the paw volume and edema in Wistar rats stimulated through λ-carrageenan-treatment. Furthermore, DKV-O treatment significantly reduced the ear edema and enhanced biopsy weight, epidermal thickness, inflammatory lesions and influx of neutrophils stimulated by TPA-treatment in CD-1 mice. DKV alone ameliorated the LPS-stimulated release of Interleukin (IL)-6, IL-17A, IL-23, and Tumor Necrosis Factor-alpha cytokines in the THP-1 cells.Taken together, DKV-O showed good efficacy in ameliorating acute systemic inflammation stimulated by effectors such as, λ-carrageenan and TPA in animal models. Hence, Divya-Kayakalp-Vati and Divya-Kayakalp-Oil co-treatment can be further explored as an anti-inflammatory treatment against dermal diseases like psoriasis and atopic dermatitis.

Sinomenine increases adenosine A2A receptor and inhibits NF-κB to inhibit arthritis in adjuvant-induced-arthritis rats and fibroblast-like synoviocytes through α7nAChR.

Sinomenine (SIN) is a clinical drug for treating rheumatoid arthritis (RA) in China. Our previous study found SIN inhibited inflammation via alpha7 nicotinic acetylcholine receptor (α7nAChR) in macrophages in vitro. Adenosine receptor A2A has anti-inflammatory and immunosuppressive function. However, the mechanisms of SIN acting on α7nAChR and the effect on adenosine A2A receptor (A2A R) in RA are not clear. In the present study, the effects of SIN on adjuvant-induced-arthritis (AIA) rats in vivo and on fibroblast-like synoviocytes (FLSs) in vitro were investigated. Indomethacin (Indo) and methotrexate (MTX), the clinical anti-arthritis drugs, were used as controls. Nicotine (Nic), a specific agonist of α7nAChR, was used as a control for targeting α7nAChR. Alpha-bungarotoxin (α-BTX), the antagonist of α7nAChR or small interference RNA (siRNA) was used to block or knock down α7nAChR. Results showed that SIN decreased arthritis index, hind paw volume, erythrocyte sedimentation (ESR) and serum TNF-α in AIA rats, and α-BTX attenuated the earlier-mentioned effects of SIN and Nic, but not Indo and MTX. The expressions of A2A R in synovium declined in AIA rats, but remarkably increased after the intervention of SIN. The expression of A2A R decreased by LPS or TNF-α, but increased by SIN; cAMP also increased by SIN in FLSs in vitro. SIN inhibited the expression of MCP-1, IL-6, and vascular endothelial growth factor in LPS-induced FLSs. SIN inhibited the activation of NF-κB. Meanwhile, α-BTX or α7nAChR siRNA blocked the earlier-mentioned effects of SIN in FLSs. Results suggested the expressions of A2A R in synovium and FLSs are negatively correlated with the arthritis progression of AIA rats and the activation of FLSs. SIN increases A2A R and inhibits the activation of NF-κB pathway via α7nAChR in AIA rats and FLSs.

Regulation of the redox signaling and inflammation by Terminalia myriocarpa leaves and the predictive interactions of it's major metabolites with iNOS and NF-ĸB

Ethnopharmacological relevanceThe present study was designed to investigate the regulation of the redox signaling and inflammation by ethanolic leaf extract of Terminalia myriocarpaVan Heurck & Müller (ETM), inspired by the reported antioxidant potential of the plant bark and the anti-edema effect of the same genus. Materials and methodsHPLC-DAD dereplication study was conducted to detect the major polyphenolic secondary metabolites. In-vitro DPPH free radical scavenging assay, nitric oxide (NO) scavenging assay, Fe2+ ion chelating ability assay and reducing power assay were conducted to evaluate the antioxidant capacity. The molecular mechanism of anti-inflammation was investigated via assessing the NO and NF-ĸB inhibiting properties in different cell lines. In-vivo carrageenan and histamine-induced edema tests were conducted using established animal models. Pro-inflammatory proteins iNOS and NF-κB were docked against the major metabolites of ETM in the in-silico study. ResultsHPLC dereplication analysis revealed the presence of considerable amount of ellagic acid, where methyl-(S)-flavogallonate was previously reported in T. myriocarpa. Significant antioxidant activity was found in every in- vitro redox assay conducted. NO was reduced in RAW 264.7 cells, showing 83.67 ± 4.18% inhibitory activity at the highest tested concentration. TNF-α induced NF-κB was also observed to be reduced in 293/NF-кB-luc cells with an inhibitory activity of 66.23 ± 0.81% at the highest dose tested. In-vivo carrageenan-induced edema test demonstrated significant anti-inflammatory activity (p < 0.05; p < 0.01) at both doses of 250 and 500 mg/kg with 60.10% highest reduction in rat paw volume. Using same doses, histamine-induced edema test exhibited mentionable anti-inflammatory potential (p < 0.05; p < 0.01) with 67.91% highest reduction in rat paw volume. Moreover, ellagic acid and methyl-(S)-flavogallonate showed significant binding affinity with iNOS (−8.5 and −8.7 Kcal/moL, respectively) and NF-κB (−7.3 and −7.3 Kcal/moL, respectively). ConclusionMentionable basis was found on behalf of the anti-inflammatory and antioxidant potentials of ETM which might be correlated with its NF-ĸB inhibiting properties.

Leaves of Stereospermum suaveolens DC Exhibit Anti-inflammatory and Anti-arthritic Potential Action in Experimental Animals

Aim: The experimental investigation of current research work was to identify traditional rich claim of Stereospermum suaveolens DC leaves for anti-inflammatory and anti-arthritic potential action in animals.&#x0D; Study design: Ethyl acetate fraction of Stereospermum suaveolens DC (Bignoniaceae) methanolic extract of leaves evaluated at 125mg/kg, 250mg/kg and 500mg/kg (p.o.) doses for anti-inflammatory and anti-arthritic activity.&#x0D; Methodology: Ethyl acetate fraction of Stereospermum suaveolens DC (Bignoniaceae) methanolic extract of leaves was evaluated for phytochemical investigation for total flavonoid content using UV spectroscopy and TLC study. Carrageenan induced rat paw edema (Acute method) and Freund’s complete adjuvant (FCA) induced chronic arthritis in wistar rats were used as an animal models to claim Stereospermum suaveolens DC leaves for anti-inflammatory and anti-arthritic potential. The rat paw volume and percentage inhibition of the paw edema were evaluated for anti-inflammatory activity. The assessments of arthritis in rats were measured by haematological values and radiological examinations.&#x0D; Result: Ethyl acetate fraction of Stereospermum suaveolens DC (Bignoniaceae) methanolic extract of leaves showed presence of total flavonoids and saponins. The significant inhibition in paw volume and edema (p &lt; .01) obtained at 250mg/kg and 500mg/kg oral dose. These obtained results were established confirmation outcome for presence of rich flavonoid contents in Stereospermum suaveolens DC leaves and provides valuable source of bioactive phytocomponents. &#x0D; Conclusion: Ethyl acetate fraction of Stereospermum suaveolens DC (Bignoniaceae) methanolic extract of leaves showed significant inhibition of inflammatory reaction as compared to standard drug indomethacin Sterospermum Suaveolens DC leaves were showed potential therapeutic role in treatment of inflammation and arthritis cases.

Simiao Wan attenuates monosodium urate crystal-induced arthritis in rats through contributing to macrophage M2 polarization

Ethnopharmacological relevanceSimiao Wan (SMW) is a classical traditional Chinese medicine (TCM) prescription to empirically treat gouty arthritis (GA) in TCM clinical practice. However, the potential mechanisms of SMW on GA are not fully evaluated. Aim of studyThe aim of this study is to investigate the role of macrophage polarization in the anti-GA activity of SMW. Materials and methodsRats were intragastricly treated with SMW for consecutive 7 days. On day 6, monosodium urate (MSU) crystal-induced arthritis (MIA) in the ankle joint was prepared. Paw volume, gait score and histological score were measured. Levels of interleukin (IL)-1β and IL-10 in serum were detected by enzyme-linked immunosorbent assay. Expressions of inducible nitric oxide synthase (iNOS), arginase (Arg)-1, phosphorylated (p)-p65, inhibitor of nuclear factor (NF)-κB (IκB)α, p-signal transducer and transcription activator (STAT)3 and p-Janus kinase (JAK)2 in synovial tissues were determined by Western blot. ResultsThe elevated paw volume, gait score and histological score in MIA rats were significantly decreased by SMW treatment. Meanwhile, SMW significantly decreased the IL-1β level and increased the IL-10 level in serum of MIA rats. Furthermore, SMW reduced the expressions of iNOS, p-p65 and enhanced the expressions of Arg-1, IκBα, p-STAT3 and p-JAK2 in synovial tissues of MIA rats. ConclusionsThe results suggest that SMW attenuates the inflammation in MIA rats through promoting macrophage M2 polarization.

MiR-223-3p inhibits inflammation and pyroptosis in monosodium urate-induced rats and fibroblast-like synoviocytes by targeting NLRP3.

Down-regulated miR-223-3p was found in rheumatoid arthritis. This study aimed to further explore the level and role of miR-223-3p in gout arthritis (GA). After monosodium urate (MSU)-induced GA rat and fibroblast-like synoviocytes (FLSs) models were established, the rat paw volume and gait score were documented and the FLSs were transfected with miR-223-3p mimic/inhibitor or NLR family pyrin domain containing 3 (NLRP3) over-expression plasmids. The MiR-223-3p target was found through bioinformatics and the dual-luciferase reporter. The rat joint pathological damage was observed by hematoxylin and eosin staining. The levels of interleukin (IL)-1β, tumor necrosis factor (TNF)-α and articular elastase in rats were detected by enzyme-linked immunosorbent assay (ELISA). The viability and pyroptosis of FLSs were detected by methyl thiazolyl tetrazolium (MTT) and flow cytometry. The expressions of miR-223-3p, NLRP3, cleaved caspase-1, IL-1β, apoptosis-associated speck-like protein (AS) and cleaved N-terminal gasdermin D (GSDMD) in FLSs or rat synovial tissues were detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR), immunofluorescence, Western blot or immunohistochemistry analysis. MSU increased the paw volume, gait score, inflammation in synovial tissues and increased the levels of IL-1β, TNF-α and articular elastase in rats. MSU decreased the viability and increased the pyroptosis of FLSs, up-regulated the expression of NLRP3, ASC, cleaved caspase-1, cleaved N-terminal GSDM, and IL-1β, and down-regulated miR-223-3p expression in synovial tissues of rat joints and FLSs. MiR-223-3p mimic reversed the effect of MSU on lowering cell viability, increasing pyroptosis in FLSs, while miR-223-3p inhibitor further enhanced the effect of MSU on FLSs. NLRP3 was a target of miR-223-3p. Also, NLRP3 over-expression reversed the effects of miR-223-3p on MSU-induced FLSs. MiR-223-3p inhibited pyroptosis in MSU-induced rats and FLSs by targeting NLRP3.

Evaluation of anti-diabetic and anti-inflammatory activities of Fenugreek (Trigonella foenum-graecum) seed extracts on Albino Wistar Rats

Background: Fenugreek (Trigonella Foenum-Graecum) is a compound used since ancient times in Indian folk medicine for several medicinal properties. It has been known to produce hypoglycemic and antioxidant actions. Aim and Objective: This study aimed to determine ethanolic extracts out of fenugreek seeds for the anti-diabetic including anti-inflammatory effects on Wister rats. Materials and Methods: Inbred Albino male and female Wistar rats were used for the study. The powdered seeds were extracted with 90% ethanol by Soxhelation (100 g) for 3–4 days. The diabetes was assessed after 72 h of alloxan-induced to rats by determining the blood sugar level. Rat Paw volume was measured to the ankle joint in drug-treated and untreated groups at 0 min, 30 min, 60 min, and 120 mins using carrageenan challenge measured mercury plethysmograph. Results: Fenugreek at 200 mg/kg dose and 400 mg/kg decreased blood glucose which was dose-dependent. The reductions in blood glucose levels (BGLs) were significant post 14th day in both groups. The antiinflammatory activity Fenugreek 200 mg/kg did not significantly reduce paw volume. Fenugreek at the dose of 400 mg/kg demonstrated inhibition of paw volume to 39.076% at the end of 2 h which was lesser than standard drug Aspirin. Conclusion: T. foenum-graecum decreased blood glucose towards the end of 21 days reduced the BGLs like the standard drug Gliclazide. The anti-inflammatory actions of the extracts were not found to be significant at the dose of 200 mg/kg and moderate anti-inflammatory actions at the dose of 400 mg/kg compared to standard drug Aspirin.

Synergistic Effects of BTK Inhibitor HM71224 and Methotrexate in a Collagen-induced Arthritis Rat Model.

Using a rat model of collagen-induced arthritis (CIA), we evaluated the therapeutic effects of HM71224 (BTKi), as well as the drug-drug interactions in combined therapy with methotrexate (MTX) based on both drugs' pharmacological role in immune regulation and antiinflammation. Arthritis in rats was induced using type II collagen and incomplete Freund's adjuvant. The therapeutic effects of HM71224 (alone or in combination with MTX) were evaluated by arthritis score, paw volume, body weight, and histopathological examination (H&E and Safranin-O staining). The drug-drug interactions between HM71224 and MTX were investigated by measuring plasma, liver enzyme and creatinine levels and blood cell counts. HM71224 reduced the clinical signs of arthritis, paw volume, and body weight loss in CIA rats. ED50 and ED90 were 1.0 and 2.5 mg/kg, respectively. HM71224 combined with MTX decreased the arthritis score, bone erosion, synovitis, and cartilage degradation without apparent interaction. The combination of HM71224 and MTX improved the therapeutic effect with no drug-drug interactions in RA.

Antioxidant, anti-inflammatory, analgesic and anti-proliferating activities of Grewia heterotricha Mast.

Introduction and Aim: Plants are considered to be novel source of active compounds having pharmacological properties and help in the development of therapeutic agents. Hence, this study was undertaken to evaluate the antioxidant, anti-inflammatory, analgesic and anti-proliferating activity of aqueous and methanolic leaf extracts of Grewia heterotricha Mast.&#x0D; Materials and Methods: The aqueous and methanolic leaf extracts of the plant were assessed for their in-vitro antioxidant activity by DPPH radical scavenging activity, in-vivo anti-inflammatory activity by carrageenan induced rat paw edema method, in-vivo analgesic activity by acetic acid-induced writhing test and in-vitro anti- proliferating activity by MTT assay.&#x0D; Results: The methanolic extract had shown very significant DPPH radical scavenging activity with IC50 value 98.95?g/ml than aqueous extract and showed a significant reduction in the paw volume of rats at the concentration of 100 mg/kg body weight indicating potent anti-inflammatory activity compared with the reference standard Diclofenac sodium. Both the extracts showed significant analgesic effect (p&lt;0.001) in acetic acid-induced pain models in a dose dependent manner. The methanolic extract showed higher analgesic activity compared to aqueous extract by inhibiting the pain indicated by a decrease in the number of writhes. In addition, both the extracts showed a decrease in MCF-7 cell viability at the concentration of 550µg/ml. Compared to the aqueous extract, MEGH has shown more cytotoxic effect on the cancer cell lines.&#x0D; Conclusion: The results suggest that both aqueous and methanolic extracts of Grewia heterotricha Mast. leaves possess potent antioxidant, analgesic, anti-inflammatory and anti-proliferating properties, which supports the use of the plant in traditional medicine. Further investigation is required to illuminate on its active compounds.&#x0D; &#x0D; Keywords: Analgesic; anti-inflammatory; DPPH; cytotoxic.

Clitoria ternatea anthocyanin extract suppresses inflammation in carrageenaninduced rat paw edema via down-regulating genes of phosphoinositide 3-kinase signaling pathway

Inflammation is characterized by an increase in vascular permeability leading to edema formation. Phosphoinositide 3-kinase (PI3K) signaling pathway plays the main role to regulate several key events in the inflammatory response to damage and infection. Our previous in silico study showed that anthocyanin and ternatin flavonoids may offer antiinflammatory potential via molecular docking model. Here, we explored whether Clitoria ternatea anthocyanin extract (CTA) had protective activity against inflammation via regulating gene expression related to PI3Ks in rat edema. CTA was extracted in methanol and acetone using maceration and lyophilization. Edema model was done by injecting carrageenan into rat paw. Oral supplementation of CTA (100, 250, and 500 mg/kg BW) and ibuprofen standard (15 mg/kg BW) was given to rats daily for 7 days. After sample treatment, the right hind paw of the rat was injected with carrageenan to induce edema condition. The ability of CTA to suppress the inflammation in paw edema was measured by quantifying the increased percentage of rat paw volume at 1, 3, and 5 hrs after injection. Rat paw tissue was collected, and gene expression related to PI3Ks, i.e. Akt1, PKB, Ilk, Pdk2, Pik3Ca, and p53 was determined using quantitative PCR (qPCR). Carrageenan showed a time-dependent increase in volume of rat paw starting from injection until the next 5 hrs. It reached maximum volume increase of rat paw at 3 hrs after injection. CTA treatment had a dose-dependent pattern in reducing the edema volume in carrageenan-induced paw edema. At 500 mg/kg BW, CTA significantly suppressed the inflammation in paw edema compared to that of positive control, indicating by the low percentage of paw edema volume at 1, 3, and 5 hrs after carrageenan injection. At gene level, qPCR data showed that CTA significantly down-regulated the expression of genes involved in PI3Ks, such as Akt1, PKB, Ilk, Pdk2, Pik3Ca, and p53 in paw tissues. Thus, CTA may have a protective effect against edema via suppressing genes of PI3Ks.