Paw Volume Research Articles

Enalapril attenuated CFA provoked arthritic manifestation by modulating proinflammatory and anti-inflammatory cytokine network

Enalapril with documented anti-inflammatory potential was evaluated in current investigation to explore its anti-arthritic efficacy. For anti-arthritic evaluation of enalapril, CFA-instigated arthritic model was employed after which various parameters comprising paw volume, body weight, arthritic index, hematological and biochemical parameters, radiographic analysis and level of various cytokines were estimated. Enalapril demonstrated significant (p˂0.001) anti-arthritic activity by suppressing paw volume, arthritic index while preserved CFA instigated weight loss. Likewise, enalapril also normalized the hematological and biochemical alterations, suppressed the level of proinflammatory cytokines with elevation of anti-inflammatory cytokines. Radiographic and histopathological analysis also further validates the anti-arthritic attribute of enalapril where enalapril preserved the normal architecture of arthritis induced joints. Outcomes of the study pointed out a notable anti-arthritic activity of enalapril. However detailed mechanistic studies are still required to point out the exact mechanism of action.

Platycodin D relieves rheumatoid arthritis by promoting apoptosis of mitochondria to inhibit activation of hedgehog pathway

Rheumatoid arthritis (RA) displays very similar characteristics to those of tumor cells, platycodin D (PD) is a triterpenoid saponin abundant in Platycodon grandiflorum (PG), plays an important role in the inhibition of tumor growth. Our previous experiments confirmed that PD inhibited MH7A cell proliferation and migration, but it’s possible mechanism remain unclear. This study aimed to reveal the mechanism of PD on RA, based on network pharmacology analysis. Rat of CIA was treated with the different doses PD. The arthritis score and paw volume were evaluated, ankle imaging changes were observed via myosseous ultrasound, all rats were anaesthetized by intraperitoneal injection of 25% urethane (1 mL/100 g), and ankle histopathology was observed using hematoxylin and eosin (HE) staining. Cell (MH7A) Counting Kit 8 (CCK8) was used to measure cell activity, and JC-1 assay kit and flow cytometry were employed to examine the cell mitochondrial membrane potential and apoptosis. The expression levels of Sonic hedgehog (Shh) signaling pathway-related proteins were observed by Western blotting. Cell inflammation levels of tumor necrosis factor alpha (TNF-α) and interleukin (IL)-6 being determined via enzyme-linked immunoassay ELISA and q-PCR. In total, the saponin PD significantly improves joint synovium inflammation and apoptosis in CIA rats. The activity of administered MH7A was significantly inhibited, the mitochondrial membrane potential decreased, the expression level of the Shh signaling pathway-related protein SuFu increased, the expression levels of SHh and Gli decreased, and cell serum levels of TNF-a and IL-6 decreased significantly. Therefore, PD exhibits therapeutic potential for synovial hyperplasia in RA.

Effect of hydroalcoholic leaf extract of Cassia fistula L. on type II collagen-induced arthritis in rats

Objective: To explore the effect of Cassia fistula on collagen II-induced arthritis in rats. Methods: The effect of 250 and 500 mg/kg chloroform and hydroalcoholic extract of Cassia fistula leaf on collagen II-induced arthritis was investigated by evaluating paw volume, arthritis index, spleen index, and biochemical parameters. Histopathological analysis and docking study were also performed. Results: A dose-dependent reduction in paw volume, arthritic index, and spleen index was observed following oral administration of the chloroform and hydroalcoholic extracts. Treatment with Cassia fistula extracts reduced tumor necrosis factor-α, interleukin (IL)-1β, IL-6, prostaglandin E2, aspartate aminotransferase, alanine aminotransferase, total leucocyte count, and erythrocyte sedimentation rate while increasing IL-10 level. In addition, Cassia fistula extracts improved joint architecture, and prevented cartilage and bone destruction. Docking analysis demonstrated that the physcion, 1-octacosanol, 5,3',4'-trihydroxy-6-methoxy-7-O-α-L-rhamnopyranosyl-(1,2)-O-β-D-galactopyranoside and scopoletin may be responsible for the anti-arthritic effect of Cassia fistula. Conclusions: Cassia fistula suppresses the progression of collagen II-induced arthritis by lowering the inflammatory factors, decreasing paw volume and arthritic index, and alleviating joint architecture. However, further studies are required to confirm the bioactive molecule responsible for the anti-arthritic potential of Cassia fistula.

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.

Anti-rheumatoid arthritis potential of Rhododendron molle G. Don leaf extract in adjuvant induced arthritis rats.

The aim of this study was to test the anti-rheumatic arthritis effects of Rhododendron molle G. Don leaf extract in arthritis rats and inflammatory RAW 264.7cells. Preliminary analysis and comparison of potential medicinal components of three polar extracts by HPLC and UHPLC-Q-TOF-MS. SD rats were subcutaneously injected with complete Freund's adjuvant (CFA) to induce inflammation on the right hind paw. RAW 264.7cells were induced by lipopolysaccharide (LPS) to established cell inflammatory model. The volume of rat hind paw was measured with a volume meter to detect swelling, and the weight of rats was measured with an electronic balance. The severity of arthritis in rats was evaluated by arthritis score. The pathological sections of rat hind paw joints were observed by hematoxylin-eosin staining, and the contents of IL-6 and IL-1β in serum were detected. qRT-PCR was used to detect the expression of IL-1β, IL-6, TNF-α and COX-2 genes in RAW 264.7cells. The release of nitric oxide was measured by Griess reaction. The expression levels of IL-6 and IL-1β were detected by Western-Blot. and discussion: The chloroform extract from R. molle leaves (CERL), Ethyl acetate extract from R. molle leaves (EERL), n-butanol extract from R. molle leaves (BERL) could significantly inhibit hind paws swelling and reduce arthritis index in arthritis rats. And it showed dose dependence. Compared with tripterygium glycosides (TG) tablets, an effective drug of RA treatment, CERL have better anti-RA effect after administration. In addition, the three kinds of the polar extracts of Rhododendron molle leaves (PERL) had lower toxicity, with the LD50 279.87, 239.65, 500.08 (mg/kg) respectively, while TG group's LD50 was 96.00 (mg/kg). In vitro experiments showed that the three PERLs can significantly inhibit the level of pro-inflammatory factors and inflammatory mediator, such as TNF-α, IL-1β, IL-6, COX-2 and NO, which were consistent with their anti-RA ability. Among the three kinds of PERLs, CERL showed the best inhibitory activity. The R. molle leaf is a potential medicinal part for the treatment of RA. This study explored the anti-RA and anti-inflammatory activities of CERL, EERL, BERL, which laid a foundation for further promoting the clinical application of R. molle.

Evaluation of Anti-Inflammatory Activity of Gum Resin Extract of Boswellia Serrata

The present investigations of gum resin extract of Boswellia Serrata were studied against experimentally anti-inflammatory activity studies. The work demonstrates that ethanolic gum resin extract and aqueous gum resin extract of Boswellia Serrata has anti-inflammation activity in mice and rat by carrageenan-induced inflammation and cotton pellet-induced granuloma. From the above observations we can conclude that ethanolic extracts and aqueous fum resin extract of Boswellia Serrata anti-inflammatory activity at both the dose level which is comparable with the standard. The ethanolic gum resin extract of Boswellia Serrata (200mg/kg), markedly increased the percentage of average mean increased in paw volume and weight in cotton pellet by the animals. The anti-inflammatory effect of both the doses (150-200 mg/kg in mice and 200-250 mg/kg in rat) showed significant activity and being that (200 mg/kg) showed higher activity. The inflammatory effects of ethanolic extract and aqueous gum resin extract of Boswellia Serrata may be attributed to any of or combination of chemicals present in the extract. Further studies are required to identify the active phytoconstituents responsible for the observed inflammatory effect of ethanol extract and aqueous extract. It is suggested and assumed that a further exploration of the present research work is needed to come up with an active anti-inflammatory.

Studies on Anti-Inflammatory Effect of Ethanolic and Aqueous Extracts of Flowers of Cassia fistula Linn. in Mice and Rat

The investigations of Cassia fistula flowers extracts were studied against experimentally anti-inflammatory activity studies and work demonstrates that ethanolic extract and aqueous extract of flowers of Cassia fistula has anti-inflammation activity in mice and rat by carrageenan-induced inflammation and cotton pellet-induced granuloma. From the above observations we can conclude that ethanolic extracts and aqueous extracts flowers of Cassia fistula anti-inflammatory activity at both the dose level which is comparable with the standard. The ethanol extract of Cassia fistula (200mg/kg), markedly increased the percentage of average mean increased in paw volume and weight in cotton pellet by the animals. The anti-inflammatory effect of both the doses (150-200 mg/kg in mice and 200-250 mg/kg in rat) showed significant activity and being that (200 mg/kg) showed higher activity. The inflammatory effects of ethanolic extract and aqueous extract of Cassia Fistula may be attributed to any of or combination of chemicals present in the extract. Further studies are required to identify the active phytoconstituents responsible for the observed inflammatory effect of ethanol extract and aqueous extract. It is suggested and assumed that a further exploration of the present research work is needed to come up with an active anti-inflammatory.

Antinociceptive and anti-inflammatory activity of DW-1021, the ionic complex of pelubiprofen and tramadol, in rodents

Although drugs such as acetaminophen, opioids, and nonsteroidal anti-inflammatory drugs (NSAIDs), are commonly used for pain management, the side effects of these drugs such as hepatotoxicity, nephrotoxicity, nausea, and vomiting, can not be neglected. Therefore, combinations of analgesics with different mechanisms raise the possibility of developing novel analgesics. Therefore, the aim of the present study was to evaluate whether DW-1021, the ionic complex of pelubiprofen (NSAID) and tramadol (opioid), has synergic antinociceptive and anti-inflammatory effects in nociceptive as well as inflammation-induced nociceptive models compared to pelubiprofen- or tramadol-only administration. Strong synergistic antinociceptive efficacy of DW-1021 was observed in the mouse writhing test and von Frey paw withdrawal threshold test in the carrageenan-induced rats. The hot plate test in mice and the Randall-Selitto mechanical paw pressure test in carrageenan-induced rats revealed that DW-1021 had a preferable effect on relieving pain to pelubiprofen, but not as much as tramadol. In the carrageenan-induced rats, DW-1021 had a more potent effect on reducing paw inflammation (paw volume, width, and thickness) via the suppression of PGE2 production than tramadol, but less than that of pelubiprofen. Taken together, our results suggest that the administration of DW-1021, a combination of pelubiprofen and tramadol, exerted a potent effect and can be used as a potential therapeutic agent for relieving pain and inflammation.

L-carnitine and Zinc supplementation impedes intestinal damage in methotrexate-treated adjuvant-induced arthritis rats: Reinstating enterocyte proliferation and trace elements

BackgroundMethotrexate (MTX), a folic acid analogue, is used as a first-line treatment for rheumatoid arthritis (RA) since it has more therapeutic mechanisms than any other drug. Being an undeniable drug for the treatment of arthritis, even low-dose MTX provokes intestinal toxicity as a primary adverse effect and does not revive an anti-inflammatory element. Thus, our study aims to elucidate the anti-arthritic and prophylactic activity of supplements L-carnitine (L) and zinc (Z) against MTX-mediated intestinal damage in arthritis rats. MethodsThe rats were assessed for arthritic parameters such as body weight, paw volume, x-ray scan, and serum trace elements level. To analyze the toxic effects of MTX in the rats, intestine pH, mucosal weight, digestive enzymes, myeloperoxidase, histopathological, and immunohistochemical analysis were performed. ResultsOur study demonstrated that the arthritic parameters have shown that MTX has an ameliorative effect on arthritic rats. Besides, our findings showed that low-dose MTX (2.5 mg/kg b.w.) given once a week for two weeks during arthritis treatment had toxic effects in the rat's intestine, as evidenced by changes in intestine pH and mucosal weight, decreased digestive enzymes, increased MPO, and degenerative changes in histopathological analysis. Concurrent therapy of LZ with MTX, on the other hand, restored the modifications in these parameters. ConclusionMTX in combination with LZ effectively manages arthritis than monotherapy and significantly prevents MTX-induced intestinal damage in arthritis rats. Thus, LZ could be used as an improved therapeutic and safety for MTX-instigated intestinal damage during arthritis treatments. Therefore, our combination of L-carnitine and zinc with MTX would be promising prophylactic activity for arthritis patients.

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.

ANTI-ARTHRITIC ACTIVITY OF ETHANOLIC EXTRACT OF CITRUS AURANTIUM LINN. LEAVES IN COMPLETE FREUND’S ADJUVANT INDUCED ARTHRITIS IN RATS

The present research work was carried out to evaluate the anti-arthritic activity of ethanolic extract of leaves of Citrus aurantium Linn. on Complete Freund’s adjuvant (CFA)-induced arthritis model in rats. Dried leaves powder was extracted and coded as ECA (ethanolic extract of C. aurantium). The antiarthritic activity of ECA was screened at the doses of 300 mg kg-1 and 500 mg kg-1. Treatment with ECA significantly decreased the paw volume, diameter, erythrocyte sedimentation rate, total white blood cell count, arthritic index and rheumatoid factor compared to arthritic rats. However, red blood cell counts and hemoglobin content were increased. The histopathological studies showed the preventive effect of ECA. This shows that ECA possesses significant anti-inflammatory and antiarthritic activity that may help treat arthritis.

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.

<i>Camphora</i> Augments Humoral Mediated Immunity and Decreases Delayed type Hypersensitivity in BALB/c Mice

Background: Immunomodulation encompasses all therapeutic interventions aimed at modifying the immune response. The immune response augments are desirable to prevent infection in immunodeficiency states and fight established diseases. In this context, the present study investigated the effect of the homeopathic medicine, Camphora, in 6CH, 30CH, and 200CH potencies on immunomodulation in experimental animals. The acute oral toxicity study was also carried out in 6CH, 30CH, and 200CH potencies to determine the safe dose volume for the immunomodulatory study. Methodology: Acute oral toxicity studies of Camphora in 6CH, 30CH, and 200CH potencies were carried out as per OECD guideline 423 with slight modifications in Wistar albino rats. Humoral immunity, i.e., primary and secondary humoral responses, was assessed by measuring the hemagglutination titre of sheep red blood cells. Delayed-Type Hypersensitivity (DTH) was evaluated by measuring footpad thickness in BALB/c mice. Results: Camphora in 6CH, 30CH and 200CH potencies at a dose volume of 2000 µl/kg did not cause any mortality in the rats when administered as a single dose. Camphora in 6CH, 30CH and 200CH potencies showed augmented primary and secondary humoral responses against the SRBC antigen in BALB/c mice. However, the values were statistically non-significant except in the case of 6 CH potency (p<0.01), which showed statistically significant primary anti-SRBC antibodies. In the DTH assay, Camphora in 6CH, 30CH and 200CH potencies significantly decreased the paw volume ratio after 24 hrs of SRBC injection in the paw, thus insinuating its role in reducing cell-mediated immunity. Camphora in 6CH, 30CH and 200CH potencies also showed enhanced antibody titres and decreased paw volume compared to vehicle control, i.e. dispensing alcohol, suggesting that the effect was imminent because of Camphora. Conclusion: The study results indicate that Camphora in 6CH, 30CH, and 200 CH potencies is safe up to a dose volume of 2000 µl/kg when administered as a single dose, augments the primary and secondary humoral immunity, and decreases DTH in experimental animals. The current study’s findings suggest that Camphora might be useful as an immunomodulator in treating immune system disorders and infectious diseases and require further investigation to investigate its mechanism of action.

Viscum monoicum: A New Source of Polyphenolic Compounds Responsible for Antioxidant and Anti-Inflammatory Activities

ABSTRACT Viscum monoicum has been used as a traditional medicinal plant for the treatment of many ailments that include pain, neuropharmacological disorders as well as various types of tumors. This study was designed to investigate the polyphenolic compound, antioxidant, and anti-inflammatory activities of the ethanolic extract of Viscum monoicum (EVM). The extract was subjected to HPLC-DAD analysis for the quantification of polyphenols. In addition, DPPH, ABTS free radical scavenging, reducing power, total antioxidant capacity, phenolic, and flavonoid content was measured for antioxidant activities of EVM. Besides, the anti-inflammatory activity of EVM was studied using carrageenan and histamine-induced rat paw edema at different doses. High levels of ellagic acid and (+)-catechin hydrate (410.15 and 340.88 mg/100 g extract, respectively) were determined for the first time through HPLC in EVM. IC50 of EVM was significant in DPPH and ABTS scavenging assay compared to that of ascorbic acid. The total antioxidant capacity, total phenolic, and total flavonoid contents were estimated and found significant amount equivalent to standard. At a dose of 400 mg kg−1, a significant anti-inflammatory activity (P<0.01) was observed in rats at both carrageenan and histamine-induced edema test models with a reduction in the paw volume; in comparison to indomethacin. Therefore, the results suggest that polyphenolic compounds might be responsible for the antioxidant and anti-inflammatory activities of EVM.

Anti-arthritic potential of jatropha curcas flowers extract using complete freund’s adjuvant (CFA)-induced arthritis model

Jatropha curcas is a member of the euphorbiaceae family and has a number of therapeutic benefits. The anti-arthritic potential of Jatropha curcas flowers extracted using various solvents is the focus of the current investigation. The use of several antioxidant assays has been researched for qualitative and quantitative evaluation of isolated chemicals and antioxidant activity. By administering crude extract orally at dose concentrations of 200 mg/kg and 400 mg/kg to rats that had Complete Freund's adjuvant (CFA)-induced arthritis, it was possible to assess the anti-arthritic effectiveness of the chosen solvent, ethanolic extract. On the 28th day of treatment, paw volume analysis and haematological parameters were analysed. The results show that extract from Jatropha curcas flowers has a sizable anti-arthritic effect.

Rimonabant-Based Compounds Bearing Hydrophobic Amino Acid Derivatives as Cannabinoid Receptor Subtype 1 Ligands.

In this study, 1H-pyrazole-3-carboxylic acids related to the cannabinoid type 1 (CB1) receptor antagonist rimonabant were amidated with valine or tert-leucine, and the resulting acids were further diversified as methyl esters, amides, and N-methyl amides. In vitro receptor binding and functional assays demonstrated a wide series of activities related to the CB1 receptors (CB1Rs). Compound 34 showed a high CB1R binding affinity (K i = 6.9 nM) and agonist activity (EC50 = 46 nM; E max = 135%). Radioligand binding and [35S]GTPγS binding assays also demonstrated its selectivity and specificity to CB1Rs. Moreover, in vivo experiments revealed that 34 was slightly more effective than the CB1 agonist WIN55,212-2 in the early phase of the formalin test, indicating a short duration of the analgesic effect. Interestingly, in a mouse model of zymosan-induced hindlimb edema, 34 was able to maintain the percentage of paw volume below 75% for 24 h following subcutaneous injection. After intraperitoneal administration, 34 increased the food intake of mice, suggesting potential activity on CB1Rs.

Formulation and Characterization of Ethyl Cellulose-Based Patches Containing Curcumin-Chitosan Nanoparticles for the Possible Management of Inflammation via Skin Delivery.

Curcumin, a natural phenolic compound, exhibits poor absorption and extensive first pass metabolism after oral administration. In the present study, curcumin-chitosan nanoparticles (cur-cs-np) were prepared and incorporated into ethyl cellulose patches for the management of inflammation via skin delivery. Ionic gelation method was used for the preparation of nanoparticles. The prepared nanoparticles were evaluated for size, zetapotential, surface morphology, drug content, and % encapsulation efficiency. The nanoparticles were then incorporated into ethyl cellulose-based patches using solvent evaporation technique. ATR-FTIR was used to study/assess incompatibility between drug and excipients. The prepared patches were evaluated physiochemically. The in vitro release, ex vivo permeation, and skin drug retention studies were carried out using Franz diffusion cells and rat skin as permeable membrane. The prepared nanoparticles were spherical, with particle size in the range of 203-229 nm, zetapotential 25-36 mV, and PDI 0.27-0.29 Mw/Mn. The drug content and %EE were 53% and 59%. Nanoparticles incorporated patches are smooth, flexible, and homogenous. The in vitro release and ex vivo permeation of curcumin from nanoparticles were higher than the patches, whereas the skin retention of curcumin was significantly higher in case of patches. The developed patches deliver cur-cs-np into the skin, where nanoparticles interact with skin negative charges and hence result in higher and prolonged retention in the skin. The higher concentration of drug in the skin helps in better management of inflammation. This was shown by anti-inflammatory activity. The inflammation (volume of paw) was significantly reduced when using patches as compared to nanoparticles. It was concluded that the incorporation of cur-cs-np into ethyl cellulose-based patches results in controlled release and hence enhanced anti-inflammatory activity.

Protective effects of harmine on Monosodium Iodoacetate-induced Osteoarthritis in rats: In vitro and in vivo studies

BackgroundOsteoarthritis (OA) is a painful and debilitating disease, which is characterized by joint pain, swelling, restricted movement, and joint stiffness. It affects more than millions of people worldwide. ObjectiveIn this current work, we aimed to assess the beneficial roles of harmine on the monosodium iodoacetate (MI)-induced OA in rats. MethodologyThe in vitro studies were carried out in the LPS-induced RAW 264.7 cells and administered with 5 and 10 µM of harmine. In in vivo studies, 2.5 mg of MI diluted in 10 ml of saline (9%) was injected into the knee joints of the experimental rats to induce OA. The 25 and 50 mg/kg of harmine was treated one week before the MI injection and continued for 25 days after induction. The paw volume and arthritis score was measured after the completion of treatments. The levels of inflammatory cytokines, PGE-2, and NO in both RAW 264.7 cells and OA-induced rats using the assay kits. The MDA and antioxidants (GSH, CAT, and SOD) levels were assessed using the assay kits. The knee joint tissues were analyzed by histopathological study. ResultsThe treatment with 5 and 10 µM of harmine effectively decreased the PGE-2 and NO levels in the LPS-exposed RAW 264.7 cells. The status of IL-6 and TNF-α also diminished by the harmine in the LPS-induced RAW 264.7 cells. The paw volume and arthritis score was decreased by the harmine treatment. The levels of PGE-2, IL-6, IL-1β, and TNF-α levels were depleted and IL-10 level was augmented by the harmine treatment in the OA-induced rats. The harmine are also suppressed the MDA and elevated the GSH, SOD, and CAT in the OA-induced rats. Further, the therapeutic roles of harmine also evidenced by the findings of the histopathological analysis. ConclusionThe current research suggests that harmine is effective in attenuating OA in rats and significantly decreasing the OA-related oxidative stress and inflammation responses. The harmine also reduced the inflammatory response in the LPS-induced RAW 264.7 cells, therefore, it may be a talented agent for treating OA.

Supplementation of syringic acid-rich Phrynium pubinerve leaves imparts protection against allergic inflammatory responses by downregulating iNOS, COX-2, and NF-κB expressions

BackgroundThe present study was designed to characterize the role of ethanolic leaf extract of Phrynium pubinerve Blume (EPP) supplement in attenuating allergic inflammation, encouraged by the presence of syringic acid in it, as this phenolic acid is reportedly promising in suppressing serum immunoglobulin E (IgE) and inflammatory cytokine levels. Materials and methodsHPLC-DAD dereplication analysis was performed to determine the presence of the vital polyphenolic metabolites. The efficacy of EPP against lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 cells was evaluated by measuring its inhibitory effects on NO and ROS/RNS production. The expressions of major inflammation-associated molecules (iNOS, COX-2, NF-κB, IL-6, and TNF-α) in RAW 264.7 cells were assessed through Western blot. Physiological and behavioral changes, BMI, and different biochemical parameters in mice blood serum were investigated in the toxicological assays. Formaldehyde-induced paw edema test in mice was conducted using established animal model. TDI-induced allergic model in mice was carried out to determine different allergy-like symptoms, and differential white blood cell (WBC) counts in blood and bronchoalveolar lavage (BAL) fluid. The intermolecular interaction analysis of the identified major metabolite of EPP with H1R and iNOS was studied by molecular docking. ResultsHPLC-DAD analysis showed the presence of syringic acid (89.19 mg/100 g EPP) and a few other compounds. LPS-induced NO generation was reduced by EPP in a concentration-dependent manner, showing IC50 of 28.20 ± 0.27 μg/mL. EPP exhibited a similar inhibitory effect on ROS/RNS production with IC50 of 29.47 ± 2.19 μg/mL. Western blotting revealed that EPP significantly downregulated the expressions of iNOS, COX-2, NF-κB, IL-6, and TNF-α in RAW 264.7 cells when challenged with LPS. The toxicological assays confirmed the dosage and organ-specific safety of EPP. In the formaldehyde-induced paw edema test, EPP caused a 66.41% reduction in mice paw volume at 500 mg/kg dose. It ameliorated TDI-induced allergy-like symptoms and decreased different inflammatory WBCs in mice's blood and BAL fluid in a dose-dependent manner. Finally, syringic acid demonstrated mentionable intermolecular binding affinity towards H1R (−6.6 Kcal/moL) and iNOS (−6.7 Kcal/moL). ConclusionsCollectively, considerable scientific reasoning was obtained in favor of the suppressive potential of EPP against allergic inflammatory responses that are proposed to be exerted via the downregulation of iNOS, COX-2, and NF-κB expressions, H1R antagonism and suppression of cytokines, such as IL-6, and TNF-α.

Gallic acid regulates immune response in a mouse model of rheumatoid arthritis.

Previous studies revealed that gallic acid (GA) exerts anti-inflammation and immuno-regulatory properties. This study aims to explore the pharmacological activities of GA in collagen-induced arthritis (CIA) mouse model. Male DBA/1J mice were used to construct the CIA model. The mice were administrated with GA for 3 weeks. Clinical arthritis scores and hind paw volume were evaluated over the experimental period. qPCR and Western blot analysis were used to determine the levels of matrix metallopeptidases (MMPs) and cytokines. In addition, flow cytometry was used to measure the populations of Th17 and Treg cells. ELISAs were used to determine the cytokines in the serum and ankle joint tissues. Treatment of GA (40 and 80 mg/kg/d) reduced clinical arthritis scores and hind paw volume in the CIA mouse model. Besides, treatment of GA reduced the overexpression of MMPs and modulated the dysregulation of inflammation-related cytokines. Flow cytometry showed that treatment of GA decreased the population of Th17 cells, and increased the population of Treg cells, as supported by treatment of GA regulated the Th17/Treg-related cytokines. GA attenuates symptoms in the CIA mouse model by anti-inflammation and regulating Th17/Treg cell imbalance.