Carrageenan-induced Rat Model Research Articles

Phenolic Profile of Seedless Ziziphusmauritiana Fruits and Leaves Extracts with In Vivo Antioxidant and Anti-inflammatory Activities: Influence on Pro-Inflammatory Mediators.

The study aimed to assess the antioxidant and anti-inflammatory activities ofpolyphenol-rich extracts of seedlessvariety of Ziziphus mauritiana(SZM). Reverse Phase High Performance Liquid Chromatographyanalysis of SZM leaves and fruit extracts in ethanol revealed the presence of sixteen phenolics including chlorogenic acid, p-coumeric acid, gallic acid, kaempferol and rutin. Leaf extract showed higher total phenolic and total flavonoid contents (177.6 mg/100 g and 46.2 mg/100 g) than in fruit extract (137.8 mg/100 g and 14.1 mg/100 g). The leaf extract exhibited higher DPPH radical-scavenging activity (63.5%) than the fruit extract (58.2%). The anti-inflammatory activity was evaluated on carrageenan-induced rat model and suppression of inflammatory biomarkers (Interleukin-6, Tumor necrosis factor-α and CRP) were studied. The fruit extract exhibited inhibition (98.1%) at the doseof 500 mg/kg body weight (BW), comparable to the indomethacin (98.4%). Both extracts suppressed the inflammatory biomarkers, but pronounced results showed by the fruit extract including CRP, IL-6, and TNF-α. The leaf extract demonstrated the higher antioxidant potential as evident from the superoxide dismutase, catalase, malondialdehyde, glutathione peroxidase and glutathione levels. These findings suggest that SZM leaf and fruit extracts possess potential antioxidant and anti-inflammatory properties and can play a significant role in mitigating oxidative stress.

Ammopiptanthus nanus (M. Pop.) Cheng f. stem ethanolic extract ameliorates rheumatoid arthritis by inhibiting PI3K/AKT/NF-κB pathway-mediated macrophage infiltration

Ethnopharmacological relevanceAmmopiptanthus nanus (M. Pop.) Cheng f. (A. nanus), a traditional Kirgiz medicinal plant, its stem has shown potential in treating rheumatoid arthritis (RA) in China, either through oral medication or by topical application directly to the affected joints, but its underlying mechanism of action remains unexplored. Aim of the studyThe purpose of this study is to elucidate pharmacological mechanism of A. nanus in ameliorating RA using a comprehensive approach that combines network pharmacology, molecular docking and experimental evaluations. Materials and methodsFirstly, the major constituents of A. nanus stem ethanolic extract were identified and quantified by High-Performance Liquid Chromatography (HPLC). Disease target data from Gene Cards database was then used to define RA-associated targets. A protein-protein interaction (PPI) network was created via STRING database. The DAVID database powered gene ontology (GO) function and kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis to gain functional insights. In vitro, RAW264.7 cells were treated with A. nanus to investigate the roles of target proteins and pathways during lipopolysaccharide (LPS) - induced inflammation. Immunofluorescence assays were performed to assess the effects of A. nanus on macrophage infiltration. The key targets and signalling pathways were validated using enzyme-linked immunosorbent assay (ELISA), real-time quantitative polymerase chain reaction (RT-qPCR), molecular docking, immunohistochemical analysis, western blotting and immunofluorescence. Finally, the therapeutic potential of A. nanus in RA was evaluated in a carrageenan-induced rat model. ResultsNetwork analysis identified 31 potential targets of A. nanus associated with RA, including 10 hub targets. KEGG analysis highlighted the involvement of PI3K/AKT signaling pathway. In vivo experiments demonstrated that A. nanus treatment significantly protected against carrageenan-induced inflammatory paw tissue and attenuated macrophage infiltration. Both in vivo and in vitro experiments confirmed that A. nanus significantly downregulated the protein expression of COX-2, iNOS and IL-1β, and inhibited PI3K/AKT/NFκB pathway, which are closely linked to RA. Furthermore, molecular docking and cellular thermal shift assay revealed that licoflavanone showed a strong binding affinity with key targets. ConclusionIn summary, this study provides the first evidence of the potent anti-inflammatory activity of A. nanus in experimental RA. The mechanism of action appears to involve inactivation of the PI3K/AKT/NF-κB pathway-mediated macrophage infiltration. These findings indicate that A. nanus has significant potential as a therapeutic potential agent for RA treatment and offer novel insights for future research and drug development in this field.

Phytochemical composition analysis, antioxidant, antimitotic, and anti-inflammatory effects of leaf and stem extracts of Pistacia lentiscus L

ABSTRACT This study explored the phytochemical composition, antioxidant properties, anti-germinative effects, and anti-inflammatory actions of methanolic leaf extracts (MLE) and methanolic stem extracts (MSE) derived from Pistacia lentiscus L. species native to the eastern region of Morocco. Using Ultra-high-performance liquid chromatography (UHPLC),16 and 17 components were discovered inMLE and MSE, respectively, with Epigallocatechin being the most prevalent in both extracts. Polyphenols and flavonoids were found to be more abundant in MLE. Furthermore, DPPH assays revealed that it had a higher free radical scavenging activity (IC50 = 37.5 mg/L) compared to ascorbic acid while ferric-reducing power tests showed that MSE had a greater potency (IC50 = 86.14 mg/L) than MLE. Both MLE and MSE inhibited sorghum seed germination in a dose-dependent manner, with MSE being slightly more effective. Mitotic index determination revealed that both extracts had concentration-dependent antimitotic effects, with MSE showing similarity to colchicine, indicating its potential as an antimitotic agent. In vitro, anti-inflammatory assays showed that it had similar properties to indomethacin in protein denaturation and membrane stabilization, while MLE displayed distinct characteristics. Furthermore, in a carrageenan-induced rat model, both extracts demonstrated anti-inflammatory potential, with MSE showing a trend of slightly better efficacy. These results highlight the importance of further investigation into MSE’s potential as an anti-inflammatory and antimitotic agent.

Enhanced Anti-Inflammatory Activity of Kaempferia galanga Extract by Solid Self-Nanoemulsifying Drug Delivery System and Its Development in Fast Disintegrating Tablet

The increasing global incidence of osteoarthritis (OA) emphasizes the need for effective treatments, particularly for the elderly. Conventional OA treatments pose administration challenges, leading to patient discomfort due to difficulties in administration. Herbal formulations offer an alternative to mitigate these issues. Kaempferia galanga (KG) contains ethyl p-methoxycinnamate, a compound displaying anti-inflammatory properties that hold promise for OA treatment. This study explores the formulation of a solid self-nanoemulsifying drug delivery system (S-SNEDDS) of Kaempferia galanga extract (KGE) and evaluates its anti inflammatory efficacy, subsequently developing it into a fast disintegrating tablet (FDT) dosage form. Initially, a liquid SNEDDS (L-SNEDDS) was prepared with varying surfactant and co surfactant concentrations. The optimal formula of L-SNEDDS (FL2) using tween 80: cremophor RH 40: PEG 400 in a ratio of 1:1:2 demonstrated favorable characteristics, including transmittance of 86.93 ± 1.16%, emulsification time of 28 ± 2.65 s, particle size of 27.79 ± 2.00 nm, PDI of 0.21 ± 0.014, and zeta potential of -12.57 mV. FL2 was solidified using aerosil 200 to produce S-SNEDDS and tested for anti-inflammatory efficacy in vivo using a carrageenan induced rat model. Results showed enhanced anti-inflammatory efficacy of KGE via S-SNEDDS, marked by reduced edema volume. Afterward, FDT S-SNEDDS were prepared using the direct compression method by comparing different types of superdisintegrants. The formulation using the combination of crospovidone and croscarmellose sodium (FT2) demonstrated excellent flow properties, tablet disintegration time of 63 s, wetting time of 34.01 ± 7.87 s, friability of 0.19%, and hardness of 3.53 ± 6.16 kg/cm2. The dissolution test indicated a better dissolution profile for FT2 compared to other formulations. In conclusion, this research presents the potential of FDT S-SNEDDS as a promising drug delivery system for enhancing the therapeutic effects of Kaempferia galanga extract in treating inflammatory conditions such as osteoarthritis.

Ropinirole inhibits inflammatory cytokine production in gingival epithelial cells and suppresses alveolar bone loss in an experimental rat model of periodontitis.

The present study explored whether the dopamine 2-like receptor agonist, ropinirole, a drug used for treating Parkinson's disease, suppresses neutrophilic inflammation and alveolar bone loss in an experimental rat model of periodontitis. Periodontitis is a neutrophilic inflammatory disease caused by periodontal pathogens. An excessive T helper (Th)17 immune response is involved in the progression of periodontitis, and interleukin (IL)-17 promotes the exacerbation of inflammation and alveolar bone destruction. Recent evidence has suggested that dopamine signaling plays a key role in Th17 cell differentiation, and that dopamine 2-like receptor agonists suppress cytokine production from Th17 cells. We previously demonstrated that tannic acid, which is a dopamine 2-like receptor agonist, inhibits alveolar bone resorption in an experimental model of periodontitis. The present study used a carrageenan-induced rat model of periodontitis with or without ropinirole. Micro-computed tomography analysis was performed. Cells of the murine gingival epithelial cell line GE1 were stimulated with carrageenan and IL-17A in the presence or absence of ropinirole. The anti-inflammatory effect of ropinirole was analyzed using reverse transcription- quantitative PCR and enzyme-linked immunosorbent assay. Subsequently, in the carrageenan-induced rat model of periodontitis, alveolar bone resorption was observed in the maxillary second molar by micro-computed tomography analysis. Intriguingly, ropinirole suppressed the alveolar bone destruction. The expression levels of C-X-C motif chemokine ligand 1 (CXCL1) and IL-17 receptor A (IL-17RA) in GE1 cells were increased by carrageenan, and CXCL1 expression in GE1 cells was upregulated under IL-17A stimulation. Moreover, ropinirole inhibited CXCL1 and IL-17RA expression in GE1 cells in the presence of IL-17A and carrageenan. Finally, haloperidol promoted CXCL1 expression in GE1 cells in the presence of carrageenan. Overall, these findings suggested that ropinirole suppressed neutrophilic inflammation and alveolar bone destruction in periodontitis by inhibiting CXCL1 expression in gingival epithelial cells through the dopamine 2-like receptor. Thus, ropinirole shows promise as a drug for the treatment of periodontitis.

Anti-Inflammatory Activity of Glabralactone, a Coumarin Compound from Angelica sinensis, via Suppression of TRIF-Dependent IRF-3 Signaling and NF-κB Pathways.

The dried root of Angelica sinensis (A. sinensis) has been widely used in Chinese traditional medicine for various diseases such as inflammation, osteoarthritis, infections, mild anemia, fatigue, and high blood pressure. Searching for the secondary metabolites of A. sinensis has been mainly conducted. However, the bioactivity of coumarins in the plant remains unexplored. Therefore, this study was designed to evaluate the anti-inflammatory activity of glabralactone, a coumarin compound from A. sinensis, using in vitro and in vivo models, and to elucidate the underlying molecular mechanisms of action. Glabralactone effectively inhibited nitric oxide production in lipopolysaccharide- (LPS-) stimulated RAW264.7 macrophage cells. The downregulation of LPS-induced mRNA and protein expression of iNOS, TNF-α, IL-1β, and miR-155 was found by glabralactone. The activation of NF-κB and TRIF-dependent IRF-3 pathway was also effectively suppressed by glabralactone in LPS-stimulated macrophages. Glabralactone (5 and 10 mg/kg) exhibited an in vivo anti-inflammatory activity with the reduction of paw edema volume in carrageenan-induced rat model, and the expressions of iNOS and IL-1β proteins were suppressed by glabralactone in the paw soft tissues of the animal model. Taken together, glabralactone exhibited an anti-inflammatory activity in in vitro and in vivo models. These findings reveal that glabralactone might be one of the potential components for the anti-inflammatory activity of A. sinensis and may be prioritized in the development of a chemotherapeutic agent for the treatment of inflammatory diseases.

Anti-inflammatory activity of Sabicea brevipes Wernharm (Rubiaceae)

Over the years, medicinal plants have been employed in the treatment of inflammation and related ailments. This study evaluated the anti-inflammatory potential of the aerial parts of S. brevipes. The extracts and fractions were further evaluated for anti-inflammatory activity in carrageenan-induced rat model at varying doses (200 and 400 mg/kg doses, orally) for 5 h of treatment. The result of the phytochemical screening showed the presence of alkaloids, terpenoids, glycosides, flavonoids and tannins in the aerial parts of the plant. The in vivo anti-inflammatory study exhibited inhibition of 42% and 44%, 47% and 36%, 33% and 31%, and 43% and 42% for methanol extract n-hexane fraction, ethyl acetate fraction, and methanol fraction, at 200 and 400 mg/kg doses, respectively. The positive control (diclofenac sodium) showed an inhibition value of 51% at 5 mg/kg dose. Finally, it is concluded that S. brevipes possesses anti-inflammatory potential which validates the enthnomedicinal claim of the plant.

Evaluation of anti-inflammatory response of berberine-loaded gum nanocomplexes in carrageenan-induced acute paw edema in rats.

Berberine is a natural plant alkaloid and has been reported to possess anti-inflammatory activity. However, berberine's poor bioavailability and low solubility have limited its clinical applicability. Nanoencapsulation of berberine using a suitable carrier can be a promising strategy to improve its efficacy. Therefore, this study aimed to produce berberine-loaded gum nanocomplexes to evaluate their therapeutic effects in a carrageenan-induced rat model. Berberine-loaded gum nanocomplexes were prepared by the ionic complexation between the negative charges of the gums (tragacanth and acacia gum) using a cross-linker for loading cationic berberine and their anti-inflammatory activity was evaluated against carrageenan-induced paw edema in rats. ELISA and qRT-PCR were employed to measure the concentration and mRNA expression level of inflammatory mediators in plasma and paw tissue, respectively. Berberine nanocomplexes were characterized for particle size (219.5nm), zeta potential by the dynamic light scattering (DLS), and for entrapment efficiency (93.2%) Infrared spectroscopy affirmed the loading of berberine in gum nanocomplexes. Transmission electron microscopy of formulation showed the spherical shape of nanocomplexes and small particle size (100-150nm). Pretreatment of rats with berberine nanocomplexes significantly reduced the paw edema in inflamed rat paws, decreased the production of nitrite and TNF-α in plasma and repressed the mRNA expression levels of TNF-α and IL-1β in paw tissue in comparison to berberine per se treated rats. The obtained berberine-loaded gum nanocomplexes produced a better anti-inflammatory effect as compared to berberine alone and hence can be used as an efficient candidate in the treatment of inflammation. The schematic representation of the preparation of the preparation of berberine-loaded tragacanth/acacia gum nanocomplexes and the evaluation in vivo for anti-inflammatory effects.

Essential oil and its nanoemulsion of Araucaria heterophylla resin: Chemical characterization, anti-inflammatory, and antipyretic activities

Araucaria heterophylla tree has been used in traditional medicine for several purposes. The essential oil (EO) of the resin is poorly studied, and up to our knowledge, the EO of the resin of the Egyptian ecospecies of A. heterophylla was not explored. In this study, the resin EO of A. heterophylla was analyzed via GC–MS. The correlation of the chemical composition of the EO with those reported in other ecospecies of A. heterophylla was evaluated via Principal Components Analysis (PCA). The in vivo inflammation inhibition and antipyretic activities in a carrageenan-induced rat model were evaluated for both crude EO and its nanoemulsion. Thirty-nine compounds were characterized in the EO. Monoterpenes (66.53 %) represented the main components followed by sesquiterpenes (30.85 %). α-Pinene (44.88 %), germacrene-D (10.25 %), α-copaene (4.72 %) and sabinene (4.44 %) were identified as major constituents of the EO. The PCA analysis showed more correlation of EO of the resin from the present study with that from the leaves of both Egyptian and Australian ecospecies, while it was negatively correlated with the EO from the leaves or resin of the Indian ecospecies. These variations could be ascribed to the environmental, climatic or genetic factors. The EO (100 mg/kg) administration in the rat paw edema and rectal temperature exhibited a significant difference (P < 0.05) by 32 %, compared to indomethacin (39 %). Also, it attenuated the levels of proinflammatory cytokines (TNF)-α, IL-6, and IL-1 β by 201.25, 285.62 and 437.0 pg/ml, respectively. Further, the administration of the higher dose of EO and its emulsion (200 mg/kg) attenuated the levels of inflammatory cytokines, improved paw edema and rectal temperature in rats. While the results showed that the low dose of nanoemulsion (100 mg/kg) was the least effective, our findings shed light toward the potentiality of using either the EO or its nanoemulsion from A. heterophylla as green resources with potent an anti-inflammatory and antipyretic promising medicinal agent.

Anti-inflammatory activity of argan oil and its minor components

Argan oil is thought to be the most expensive edible oil worldwide. It is difficult to produce and the argan tree only grows in a limited geographical area, notably Morocco and Algeria. Because it is produced by mechanical means, argan oil contains “minor” components that might be endowed with healthful effects. We investigated in vivo the anti-inflammatory activities of argan oil and its unsaponifiable fraction, using diclofenac as the control, in a carrageenan-induced rat model of inflammation. Rats were given different amounts of argan oil or its unsaponifiable fraction, by gavage. We report that argan oil and its “minor” components effectively lessen the inflammatory actions of carrageenan. Far from being “pharmacological” the actions of argan oil are comparable with those of diclofenac in the short, i.e. 4 h term. Sustained consumption of argan oil might, therefore, contribute to lessen the burden of degenerative diseases associated with higher inflammatory status.

Thrombolytic Effects in vivo of Nattokinase in a Carrageenan-Induced Rat Model of Thrombosis

Background/Aims: Nattokinase is a serine protease produced by Bacillus subtilis during the fermentation of the soybean product natto. The fibrinolytic activity and thrombolytic effects of nattokinase have been observed in vitro, but the effect in vivo has still to be researched. The objective of this study was to demonstrate the activity of nattokinase in vivo. Methods: To establish a rat model of thrombosis, κ-carrageenan was injected subcutaneously into the toes of Sprague-Dawley (SD) rats. Histological examination confirmed thrombosis. The rats were then treated with varying doses of nattokinase and the resulting thrombolysis was histologically assessed. ELISA was used to determine the levels of the fibrin/fibrinogen degradation products (FDPs) and D-dimer, which are sensitive indices of fibrinolytic activity. Vermis kinase, a known thrombolytic agent, was used as a positive control. Results: Biopsy results revealed partial thrombolysis in the tail vessels of the rats treated with nattokinase or vermis kinase. FDP and D-dimer levels were higher in rats treated with high-dose nattokinase than in those treated with saline. No difference in FDP or D-dimer levels was observed between rats treated with high-dose nattokinase and those treated with vermis kinase. Conclusions: Both the histological and physiological evidence from this study indicate that nattokinase exerts thrombolytic effects in vivo.

Inhibitory Effect of the Hexane Fraction of the Ethanolic Extract of the Fruits ofPterodon pubescensBenth in Acute and Chronic Inflammation

Fruits of Pterodon pubescens Benth have been used traditionally for the treatment of rheumatism, sore throat, and respiratory disorders, and also as anti-inflammatory, analgesic, depurative, tonic, and hypoglycemic agent. The study was aimed at evaluating the anti-inflammatory activity of the hexane fraction of an ethanolic extract of P. pubescens fruits. The oil from P. pubescens fruits was extracted with ethanol and partitioned with hexane. The anti-inflammatory activity was measured with increasing doses of the hexane fraction (FHPp) by using a carrageenan-induced rat model of pleurisy and a rat model of complete Freund's adjuvant-induced arthritis by using an FHPp dose of 250 mg/kg for 21 days. Treatment with an FHPp resulted in anti-inflammatory activity in both models. The results of biochemical, hematological, and histological analyses indicated a significant decrease in glucose, cholesterol, and triglycerides levels (18.32%, 34.20%, and 41.70%, resp.) and reduction in the numbers of total leukocytes and mononuclear cells. The FHPp dose of 1000 mg/kg induced no changes in behavioral parameters, and no animal died. The results of this study extend the findings of previous reports that have shown that administration of extracts and fractions obtained from species of the genus Pterodon exhibits anti-inflammatory activity and lacks toxicity.

Electroacupuncture inhibits phosphorylation of spinal phosphatidylinositol 3-kinase/Akt in a carrageenan-induced inflammatory rat model

We investigated the changes of pain-related spinal signaling pathway after electroacupuncture (EA) stimulation in a carrageenan-induced rat model. EA stimulation (2Hz, 1mA) was needle-delivered for 30 min at acupoints corresponding to Zusanli and Sanyinjiao 3 h after carrageenan injection. Thermal and mechanical sensitivity of the hindpaw induced by carrageenan was strongly inhibited by EA stimulation. Phosphorylation of extracellular signal-regulated kinase, phosphatidylinositol 3-kinase (PI3K), Akt, and cAMP response element-binding protein (CREB) were examined in the L4-5 segments of the spinal cord by Western blot analysis 4h and 5h after carrageenan injection. Phosphorylation of Akt and especially PI3K were significantly induced by carrageenan-induction, but these expressions were markedly inhibited by EA stimulation. CREB phosphorylation showed a similar, but insignificant, pattern as like PI3k/Akt. Immunohistochemical analyses confirmed that phosphorylation of PI3K and Akt showed a similar pattern as Western blotting and were observed in most neurons and a few astrocytes. EA and PI3K inhibitor synergistically inhibited carrageenan-induced hyperalgesia. These results reveal that both neuronal PI3K and Akt may play an important role in EA-induced antinociception via inactivation in an inflammatory pain model.