Carrageenan-induced Paw Edema Model Research Articles

Design, optimization and evaluation of dexamethasone-loaded microneedles for inflammatory disorders

Dexamethasone (Dex) is a popular and highly potent anti-inflammatory drug, frequently used to treat a wide range of inflammatory disorders. However, the existing oral and parenteral delivery modes have several limitations, including systemic adverse effects and reduced patient compliance. This study aimed to develop a biodegradable microneedle (MN)-based transdermal delivery system capable of sustained, safe and effective delivery of Dex. A Quality by Design (QbD) approach was applied to design the Dex-loaded MN arrays. The formulation variables were optimized using a central composite design (CCD) model, generated with the statistical software package Design- Expert®. The optimized MNs were sharp, with heights ranging between 800 and 900 µm, appropriate for transdermal delivery. The MN arrays did not exhibit any cytotoxic effects on the fibroblast and keratinocyte cells. Moreover, the ex vivo studies confirmed the enhanced efficacy of MN-mediated skin permeation of Dex compared to passive permeation of drug solution. Finally, the in vivo anti-inflammatory efficacy was investigated using the carrageenan-induced rat paw edema model. The efficacy of the MN arrays to inhibit paw edema formation was found to be comparable to that of intravenous Dex injection and significantly greater than topical solution. Cytokine analysis also revealed that application of MN arrays downregulated the expressions of pro-inflammatory cytokines and upregulated the expressions of anti-inflammatory cytokines. Overall, the findings suggest that MN array could be a safe, easy, effective and minimally invasive alternative to the existing means of Dex delivery and could potentially be used for the treatment of inflammatory disorders.

Assessment of formulation variables of poor water soluble diacerein for its improved loading and anti-inflammatory activity.

Dissolving microneedles have become a popular method for percutaneous administrationof drugs. However, loading poorly soluble drugs into water-based dissolving microneedles remains a challenge. In view of this, we aimed to improve Diacerein (DCN) solubility formulating dissolving microneedles. DCN microsuspension was created by high-speed homogenization with organic solvents or wet milling with Tween 80 as a stabilizer (LD1). They were analyzed for particle size and saturation solubility. Subsequently, the organic solvent-based microneedles were prepared under vacuum, whereas LD1 was mixed with HPMC (8% w/w) and PVP (30% w/w) matrix to concentrate the drug in acral fraction through centrifugation. DCN microsuspension in DMSO had the highest drug solubility with an average particle size of 6 µm, whereas LD1 had a particle size of 3.28 µm showing improved solubility. TD-3 had the highest drug loading and the least amount of drug migration into the blank baseplate. Within 5 min, these microneedles dissolved completely in an agarose-gel block. LD1 was likewise put in the baseplate to generate TD3-B. Within 24 h, 74.39% of the medication was released from TD3-B, with only a small amount remaining in the baseplate. TLC examination indicated the conversion of DCN to Rhein in the skin, whereas DSC and TGA studies revealed amorphous features. DCN microneedles showed no sign of skin irritancy but showed anti-inflammatory response on carrageenan-induced paw edema model. Microneedles remained stable during accelerated stability testing. Wet milling in the presence of a stabilizer can be an effective approach for enhancing DCN solubility for improved drug loading in dissolving microneedles. Improvement in solubility of Diacerein for subsequent loading in Dissolving Microneedle for percutaneous delivery.

Anti-inflammatory Activity of the Phenolic Compounds from Vanda roxburghii R. Br.

Inflammation is implicated in the pathogenesis of several neurodegenerative diseases including Alzheimer’s disease (AD). Vanda roxburghii R. Br. is used as folk medicine in the treatment of various ailments and diseases including inflammation, rheumatoid arthritis and neurological disorders. We reported earlier that the plant possesses anti-inflammatory activity and the activity is high in the chloroform fraction among other fractions tested. Therefore, the present work was designed to isolate the compounds from the chloroform fraction responsible for anti-inflammatory activity. Four compounds namely syringaldehyde (1), vanillin (2), dihydroconiferyl dihydro-p-coumarate and (3) and gigantol (4) were isolated and purified using chromatographic methods. The isolated compounds were identified by analysis of their NMR spectral data. The compounds were evaluated for anti-inflammatory activity using carrageenan-induced hind paw edema model in mice. All the compounds showed significant activity at a dose of 100 mg/kg. Among the compounds, gigantol displayed the highest activity which was comparable to the activity of the standard drug indomethacin. At the same dose the other three compounds syringaldehyde, vanillin and dihydroconiferyl dihydro-p-coumarate exhibited similar activity. These results suggest that the isolated compounds from the chloroform fraction have potential anti-inflammatory activity that might be useful in the prevention and treatment of AD. Bangladesh Pharmaceutical Journal 26(1): 1-6, 2023 (January)

Tailoring of Novel Bile Salt Stabilized Vesicles for Enhanced Transdermal Delivery of Simvastatin: A New Therapeutic Approach against Inflammation.

Simvastatin (SMV), a cholesterol-lowering agent, has antioxidant and anti-inflammatory effects. Nevertheless, the oral use of SMV is linked with poor systemic bioavailability owing to its limited aqueous solubility and extensive first-pass metabolism. The aim of this study was to evaluate the feasibility of transdermal delivery of SMV using bile salt stabilized vesicles (bilosomes) for enhancing the anti-inflammatory potential of SMV. SMV-loaded bilosomes (SMV-BS) were prepared by the thin film hydration technique and optimized by 33 Box-Behnken design. The fabricated SMV-BS were assessed for vesicle size, entrapment efficiency (% EE) and cumulative drug release. The optimized formula was incorporated into HPMC gel and investigated for physical properties, ex vivo permeation, in vivo pharmacokinetic study and anti-inflammatory potential in inflamed paw edema rat model. The optimized SMV-BS showed vesicle size of 172.1 ± 8.1 nm and % EE of 89.2 ± 1.8%. In addition, encapsulating SMV within bilosomal vesicles remarkably sustained drug release over 12 h, compared to plain drug suspension. Furthermore, SMV-loaded bilosomal gel showed a three-fold enhancement in SMV transdermal flux, compared to plain drug suspension. Most importantly, the relative bioavailability of SMV-BS gel was ~2-fold and ~3-fold higher than those of oral SMV suspension and SMV gel, respectively. In carrageenan-induced paw edema model, SMV-BS gel induced a potent anti-inflammatory effect, as evidenced by a remarkable reduction in paw edema, which was comparable to that of the standard anti-inflammatory drug, indomethacin. Collectively, bilosomes might represent a plausible transdermal drug delivery system that could enhance the anti-inflammatory activity of SMV by boosting its skin permeation and its systemic bioavailability.

Evaluation of the anti-inflammatory, antioxidant, and cytotoxic potential of Cardamine amara L. (Brassicaceae): A comprehensive biochemical, toxicological, and in silico computational study.

Introduction: Cardamine amara L. (Brassicaceae) is an important edible plant with ethnomedicinal significance. This study aimed at evaluating the phytochemical composition, anti-inflammatory, antioxidant and cytotoxicity aspects of the hydro-alcoholic extract of C. amara (HAECA). Methods: The phytochemical composition was evaluated through total phenolic contents (TPC), total flavonoid contents (TFC) determination and UPLC-QTOF-MS profiling. Anti-inflammatory evaluation of HAECA was carried out through the carrageenan induced paw edema model. Four in vitro methods were applied in the antioxidant evaluation of HAECA. MTT assay was used to investigate the toxicity profile of the species against human normal liver cells (HL7702), human liver cancer cell lines (HepG2) and human breast cancer cell lines (MCF-7). Three major compounds (Gentisic acid, skullcapflavone and conidendrine) identified in UPLC-Q-TOF-MS analysis were selected for in silico study against cyclooxygenase (COX-I and COX-II). Results and Discussion: The findings revealed that HAECA is rich in TPC (39.32 ± 2.3mg GAE/g DE) and TFC (17.26 ± 0.8mg RE/g DE). A total of 21 secondary metabolites were tentatively identified in UPLC-Q-TOF-MS analysis. In the MTT cytotoxicity assay, the extract showed low toxicity against normal cell lines, while significant anticancer activity was observed against human liver and breast cancer cells. The carrageenan induced inflammation was inhibited by HAECA in a dose dependent manner and showed a marked alleviation in the levels of oxidative stress (catalase, SOD, GSH) and inflammatory markers (TNF-α, IL-1β). Similarly, HAECA showed maximum antioxidant activity through the Cupric reducing power antioxidant capacity (CUPRAC) assay (31.21 ± 0.3mg TE/g DE). The in silico study revealed a significant molecular docking score of the three studied compounds against COX-I and COX-I. Conclusively the current study encourages the use of C. amara as a novel polyphenolic rich source with anti-inflammatory and antioxidant potential and warrants further investigations on its toxicity profile.

PpSP32, the Phlebotomus papatasi immunodominant salivary protein, exerts immunomodulatory effects on human monocytes, macrophages, and lymphocytes

BackgroundThe saliva of sand flies, vectors of Leishmania parasites, contains several components that exert pharmacological activity facilitating the acquisition of blood by the insect and contributing to the establishment of infection. Previously, we demonstrated that PpSP32 is the immunodominant salivary antigen in humans exposed to Phlebotomus papatasi bites and validated its usefulness as a predictive biomarker of disease. PpSP32, whose functions are little known to date, is an intriguing protein due to its involvement in the etiopathogenesis of pemphigus, an auto-immune disease. Herein, we aimed to better decipher its role through the screening of several immunomodulatory activity either on lymphocytes or on monocytes/macrophages.MethodsPeripheral mononuclear cells from healthy volunteers were stimulated with anti-CD3/anti-CD28 antibodies, phytohemagglutinin, phorbol 12-myristate 13-acetate/ionomycin, or lipopolysaccharide in the presence of increasing doses of PpSP32. Cell proliferation was measured after the addition of tritiated thymidine. Monocyte activation was tested by analyzing the expression of CD86 and HLA-DR molecules by flow cytometry. Cytokine production was analyzed in culture supernatants by ELISA. THP-1-derived macrophages were stimulated with LPS in the presence of increasing doses of PpSP32, and cytokine production was analyzed in culture supernatants by ELISA and multiplex technique. The effect of PpSP32 on NF-kB signaling was tested by Western blot. The anti-inflammatory activity of PpSP32 was assessed in vivo in an experimental inflammatory model of carrageenan-induced paw edema in rats.ResultsOur data showed that PpSP32 down-modulated the expression of activation markers in LPS-stimulated monocytes and THP1-derived macrophages. This protein negatively modulated the secretion of Th1 and Th2 cytokines by human lymphocytes as well as pro-inflammatory cytokines by monocytes, and THP1-derived macrophages. PpSP32 treatment led to a dose-dependent reduction of IκB phosphorylation. When PpSP32 was injected into the paw of carrageenan-injected rats, edema was significantly reduced.ConclusionsOur data indicates that PpSP32 induces a potent immunomodulatory effect on monocytes and THP-1-derived macrophages. This inhibition could be mediated, among others, by the modulation of the NF-kB signaling pathway. The anti-inflammatory activity of PpSP32 was confirmed in vivo in the carrageenan-induced paw edema model in rats.Graphical

Ursolic Acid Derivatives Down Regulate Inflammatory Mediators

Ursolic acid (UA) is being investigated due to its anti-inflammatory potential, and structural modifications can enhance its biological activities. The aim of this study was to evaluate the immunomodulatory effect of the ursolic acid derivatives (UAD) in macrophages and in the carrageenan-induced paw edema model. RAW264.7 cells were cultured in the presence or absence of UA or UAD (1-18). Nitric oxide (NO), nuclear factor kappa B (NF-κB), tumor necrosis factor (TNF), and cellular viability were measured. 30 min before the carrageenan-induced paw edema, the UAD1 and UAD2 (200 mg kg-1) were administered intraperitoneally. The results showed that UAD2-4, UAD7, UAD9-11 had half maximal inhibitory concentration (IC50) greater than 90 µM and were able to reduce NO, NF-κB and TNF production. Moreover, UAD1 and UAD2 reduced paw edema and IL-6 production. In conclusion, the results obtained demonstrated a variation in the response between the derivatives due to the chemical modifications, showing potential to reduce the inflammatory mediators, deserving further investigations.

Synthesis and Biological Evaluation of Some New Chalcone Derivatives as Anti-inflammatory Agents.

Aim-The present research work aims to prepare a series of 1-(4-(2-(1H-indol-1-yl)-2-oxoethoxy)phenyl)-3-phenylprop-2-en-1-one derivatives. Method- The major compound was achieved by the reaction of indole with chloroacetylchloride in benzene afforded 2-chloro-1-(indoline-1-yl) ethanone which reacts o- hydroxy acetophenone in presence of acetonitrile to form 2-(4-acetylphenoxy)-1-(1H-indol-1-yl)ethan-1-one then is goes through aldol condensation to give various final derivatives. Results and conclusion- After synthesis of compounds, the synthesized compounds were characterized by checking their solubility, melting point, thin layer chromatography, IR, 1HNMR spectral data and elemental analysis. All of prepared derivatives were evaluated for their anti-inflammatory activity on wistar albino rats by following carrageenan induced Rat Hind Paw Edema model.

Analgesic and Anti-Inflammatory Effects of 80% Methanol Extract and Solvent Fractions of the Leaves of Vernonia auriculifera Hiern. (Asteraceae).

The leaves of V. auriculiferahas been used traditionally for the treatment of inflammatory disorders, and pain in various parts of Ethiopia. However, to our knowledge, the analgesic and anti-inflammatory activity of the crude extract and solvent fractions has never been experimentally studied. To assess the analgesic and anti-inflammatory activitiesof V. auriculifera leaf extract and solvent fractions in rodent models. Air-dried leaves ofV. auriculiferawere grounded and macerated using 80% methanol. The air-dried, grounded leaves were also successively extracted with ethyl acetate, and methanol. The residue was then macerated in water for 72 hr. The extract's peripheral analgesic activity, as well as the solvent fractions, were determined using an acetic acid-induced writhing test. The hot plate model was used to assess the central analgesic effect. Carrageenan-induced hind paw edema and cotton pellet-induced granuloma models were used to assess the anti-inflammatory effect in rats. The 80% methanol leaf extract and solvent fractions have demonstrated significant (p < 0.05) peripheral and central analgesic activity. Both 80% methanol leaf extract and solvent fractions of V. auriculifera were found to have anti-inflammatory activity in a carrageenan-induced rat paw edema model. In the cotton pellet-induced granuloma model, all concentrations of 80% methanol leaf extract (ME), methanol fraction (MEF), and aqueous fractions (AQF) of V. auriculifera inhibited exudate and granuloma formation. Although all tested doses significantly inhibited granuloma mass formation, only the medium and highest ethyl acetate fraction (EAF) doses significantly inhibited the generation of inflammatory exudate. This study's findings indicate that the solvent fractions and 80% methanol extract of V. auriculifera have analgesic and anti-inflammatory properties. This study's findings not only confirm the plants' traditional claim but also provide clues for further investigation of the active principles of this plant for the development of effective and safe analgesic and anti-inflammatory drugs.

Anti-inflammatory effect of ethanolic extract of Aegle marmelos leaves on carrageenan-induced rat paw edema

Background: Inflammation is an essential biological protective response of the body which can occur due to various external and internal stimuli. Although it is an essential response, excess inflammation can be detrimental and hence it should be controlled with appropriate medication. Aims and Objectives: To assess the anti-inflammatory property of ethanolic extract of Aegle marmelos leaves in Wistar rats. Materials and Methods: A total of 24 male adult Wistar albino rats weighing approximately 150–200 g were used in the study. Acute anti-inflammatory activity was tested by employing a carrageenan-induced paw edema model. 24 rats were divided randomly into 4 groups. The study was conducted with Group I as the control group. Groups II and III received indomethacin (25 mg/kg) and 200 mg/kg of extract respectively. Group IV received extract and half dose of indomethacin (12.5 mg/kg). The anti-inflammatory property was expressed in terms of percentage inhibition (PI). Statistical analysis was conducted by one-way analysis of variance, following which Tukey’s post hoc test was done for multiple comparisons. P &lt; 0.05 was considered to be a statistically significant value. Results: Mean paw edema was significantly reduced in both Groups III and IV at all hours of inflammation compared to control. Group III exhibited maximum PI of 27% at 4 h and Group IV exhibited maximum PI of 49% at 6 h of inflammation. The anti-inflammatory activity of Group IV was comparable to Group II. Conclusion: As a result of the above findings it can be concluded that the ethanolic extract of Aegle leaf has significant anti-inflammatory properties.

The dichloromethane extract of Callicarpa longissima rich in diterpenoid phenols exerts anti-inflammatory effect via inhibiting the TLR4/NF-κB signaling pathway

Ethnopharmacological relevanceCallicarpa longissima is a typical Yao ethnomedicine that has been used to treat arthritis in China. Our previous study found that the dichloromethane extract (DCME) of C. longissima showed anti-inflammatory activity in vitro. However, the anti-inflammatory mechanism and detailed chemical composition of DCME remain unclear, which lead to the original interest of this study. Aim of the studyThe study aimed to evaluate the anti-inflammatory properties of the DCME from C. longissima and further explore the accurate chemical components responsible for this active extract. Materials and methodsThe anti-inflammatory activity of DCME in vivo was tested with carrageenan-induced mice paw edema model. Its anti-inflammatory mechanism was explored with LPS-stimulated RAW264.7 macrophages model. The compounds in DCME were isolated by repeated column chromatography and their structures were identified on the basis of nuclear magnetic resonance spectroscopy. The anti-inflammatory activities of the isolates in vitro were also tested by suppressing releases of inflammatory mediators (NO, IL-6 and TNF-α) in RAW264.7 macrophages model. In addition, the molecular docking analysis, which evaluated the potential interaction between the compounds and Toll-like receptor 4 (TLR4) and nuclear factor κB (NF-κB), was performed. ResultsDCME effectively alleviated the mice paw edema induced by carrageenan. In LPS-stimulated RAW264.7 cells, DCME significantly decreased the production of interleukin (IL)-6 and tumor necrosis factor α (TNF-α) via inhibiting their mRNA transcription, down-regulated the expression of TLR4 and myeloid differentiation factor 88, inhibited the phosphorylation of alpha inhibitor of NF-κB (IκBα), NF-κB p65, and degradation of IκBα. Twelve diterpenoid phenols were identified from DCME, and they not only showed different inhibitory effects on the production of NO, IL-6 and TNF-α in LPS-stimulated RAW264.7 cells, but also could bind to TLR4 and NF-κB as analyzed by molecular docking. ConclusionsTaken together, DCME from C. longissima could inhibit inflammatory response both in vitro and in vivo, which is mainly attributed to the synergistic effect of abundant diterpenoid phenols through inhibiting the TLR4/NF-κB signaling pathway, and might be a promising agent for the treatment of inflammatory diseases.

Potential anti-inflammatory, hypoglycemic, and hypolipidemic activities of alpha-pinene in diabetic rats

Diabetes mellitus (DM) is a chronic disease characterised by high levels of glucose in the blood, resulting from a relative or absolute deficiency in the action of insulin. It is now clear that a state of chronic low- level inflammation is closely associated with the pathophysiology of DM, which may contribute to the long-term effects of the disease. Some monoterpenes, compounds present in oils from various aromatic plants, such as β- pinene, have antidiabetic and anti-inflammatory properties. Thus, the current study sought to assess the hypoglycemic, hypolipidemic, and anti-inflammatory effects of α-pinene, an isomer of β-pinene, in diabetic rats. Methods: Male Wistar rats (200–250 g) received a single injection of alloxan (40 mg/kg) through the penile vein to induce diabetes. Diabetic animals were orally treated with vehicle or α-pinene (25, 50, 100 mg/kg) or glibenclamide (5 mg/kg) for seven consecutive days. On the last day of treatment, blood samples were collected for biochemical analysis and the anti-inflammatory effect of α-pinene was evaluated using the carrageenan-induced paw edema and carrageenan-induced peritonitis models. Results: The treatment with the monoterpene decreased blood glucose, cholesterol, triglyceride, VLDL, LDL, HDL, AST and ALT levels when compared to the vehicle group. Likewise, the association α-pinene + glibenclamide also decreased blood glucose, total cholesterol and triglyceride levels. The oral treatment with α-pinene inhibited carrageenan-induced recruitment of total leukocytes to the peritoneal cavity and also inhibited carrageenan-induced paw edema. Alpha-pinene showed significant hypoglycemic and hypolipemic activity at doses 25, 50 and 100 mg/kg. The strongest anti-inflammatory activity of α-pinene was observed at a dose of 50 mg/kg. Finally, these findings suggest that α-pinene may have anti-inflammatory, hypoglycemic, and hypolipemic properties. Its anti-inflammatory properties are probably an asset for the treatment of diabetes and its complications. However, new preclinical tests are required to elucidate the mechanism of action by which α-pinene exerts its activity.

Smart redox-sensitive micelles based on chitosan for dasatinib delivery in suppressing inflammatory diseases

Dasatinib (DAS) exhibits anti-inflammatory effects by retrieving the balance between inflammatory and anti-inflammatory cytokines secreted by macrophages. The aim of this study was the development of redox-responsive micelles with the potential of passive targeting and on-demand drug release for DAS delivery to macrophages. For this purpose, two molecular weights of chitosan (CHIT) were conjugated to DAS at different molar ratios using 3,3′-dithiodipropionic anhydride (DTDPA) as disulfide bond containing linker to synthesize a series of CHIT-S-S-DAS amphiphilic conjugates. Micelles obtained by the sonication method had particle sizes of 129.3–172.2 nm, zeta potentials of +17.5 to +20.9 mV, drug contents of 0.90–7.20 %, CMC values of 35.3–96.6 μg/ml, and exhibited redox-responsive in vitro drug release. Optimized micelles were non-toxic and dramatically more efficient than non-redox responsive micelles in reducing TNF-α and IL-6 and increasing IL-10 secretion from LPS-stimulated RAW264.7 cells. Furthermore, the redox-responsive micelles were able to reduce the mice paw edema, reduce the plasma levels of pro-inflammatory cytokines and increase plasma level of IL-10, considerably more than free DAS and non-redox responsive micelles in carrageenan-induced mice paw edema model of inflammation.

Rutin Nanocrystals with Enhanced Anti-Inflammatory Activity: Preparation and Ex Vivo/In Vivo Evaluation in an Inflammatory Rat Model.

Rutin is a polyphenolic flavonoid with an interestingly wide therapeutic spectrum. However, its clinical benefits are limited by its poor aqueous solubility and low bioavailability. In an attempt to overcome these limitations, rutin nanocrystals were prepared using various stabilizers including nonionic surfactants and nonionic polymers. The nanocrystals were evaluated for particle size, zeta potential, drug entrapment efficiency, morphology, colloidal stability, rutin photostability, dissolution rate, and saturation solubility. The selected nanocrystal formulation was dispersed in a hydrogel base and the drug release kinetics and permeability through mouse skin were characterized. Rutin's anti-inflammatory efficacy was studied in a carrageenan-induced rat paw edema model. The nanocrystals had a size in the range of around 270-500 nm and a polydispersity index of around 0.3-0.5. Nanocrystals stabilized by hydroxypropyl beta-cyclodextrin (HP-β-CD) had the smallest particle size, highest drug entrapment efficiency, best colloidal stability, and highest drug photostability. Nanocrystals had around a 102- to 202-fold and 2.3- to 6.7-fold increase in the drug aqueous solubility and dissolution rate, respectively, depending on the type of stabilizer. HP-β-CD nanocrystals hydrogel had a significantly higher percent of drug released and permeated through the mouse skin compared with the free drug hydrogel. The cumulative drug amount permeated through the skin was 2.5-fold higher than that of the free drug hydrogel. In vivo studies showed that HP-β-CD-stabilized rutin nanocrystals hydrogel had significantly higher edema inhibition compared with the free drug hydrogel and commercial diclofenac sodium gel. These results highlight the potential of HP-β-CD-stabilized nanocrystals as a promising approach to enhance drug solubility, dissolution rate, and anti-inflammatory properties.

Phenolic compounds and extracts from Crotalaria calycina Schrank potentially alleviate pain and inflammation through inhibition of cyclooxygenase-2: An in vivo and molecular dynamics studies

Crotalaria calycina Schrank is a local Bangladeshi plant well-accepted by the tribal population for its medicinal properties. The primary approach of our study was to uncover the analgesic and anti-inflammatory potential of methanol extract of C. calycina stem in mice model with in silico molecular docking and molecular dynamics simulation approach. Phenolic compounds were identified and quantified from the extract through high-performance liquid chromatography-diode array detector (HPLC-DAD) analysis. Writhing assay through injection of acetic acid, licking assay through formalin injection, and finally, hot plate assay was employed to observe the analgesic activity. The carrageenan-induced paw edema model was employed to determine the anti-inflammatory potential of the extract. In silico molecular docking and molecular dynamics were also run to validate the in vivo study results. Eight polyphenolic compounds from the extract were identified and quantified via HPLC-DAD analysis, and (-) epicatechin was most abundantly distributed (87.15 ± 0.24 mg/100 g dry extract). In vivo study revealed that 400 mg/kg dose significantly inhibited (P < 0.01) the writhing response in the writhing assay and demonstrated the highest percent of inhibition of licking (70.67%) in the late part of the licking test. The same extract dose produced the highest (74.71%) percent of maximal effect (% MPE) in the hot plate assay. It demonstrated the highest percent of edema inhibition (68.00%) in the fourth hour of the paw edema assay. Molecular docking and molecular dynamics simulation of (-) epicatechin, caffeic acid, and kaempferol with cyclooxygenase-2 revealed that they have similar interactions to the standard inhibitor celecoxib. These valuable bioactive compounds may induce significant analgesic and anti-inflammatory properties in MECCS. Therefore, based on the findings of this study, it can be concluded that C. calycina stem can be a prospect in the medicinal field due to its remarkable analgesic and anti-inflammatory effect.

Screening of aqueous extract of fenugreek seeds for its anti-inflammatory potential in albino rats

Background: Inflammation is a common defense mechanism that involves a complicated network of cell-cell, cell-mediator, and tissue interaction. Fenugreek (Trigonella foenumgraecum L.) seeds are used for the treatment of many inflammatory conditions. Aims and Objectives: The aim of the study was to analyze the anti-inflammatory properties of aqueous extracts of fenugreek seeds in albino rats and compare it to control and standard anti-inflammatory treatments in animal models of acute and chronic inflammation. Materials and Methods: The methods employed to study the anti-inflammatory activity are carrageenan-induced rat paw edema and turpentine induced arthritis model in rats for acute inflammatory model and cotton wool pellet granuloma in rats for chronic inflammatory modelfollowing acquisition of the Institutional Ethics Committee approval. The control group was given 1% gum acacia, standard group was given 10 mg/kg Indomethacin dissolved in distilled water and test group was given 200 mg/kg of aqueous extract of fenugreek seeds suspended in 1% gum acacia orally. Results: The test group displayed almost equal increase in paw volume edema as that of the standard group under experimental conditions. In Carrageenan-induced rat paw edema model and turpentine induced arthritis model, aqueous extract of fenugreek seeds and indomethacin had comparable anti-inflammatory effect. The anti-granuloma effect of aqueous extract of fenugreek seeds was of moderate degree compared to the standard under the present experimental conditions. Conclusion: When compared to standard indomethacin, the aqueous extract of fenugreek seeds demonstrated significant anti-inflammatory activity in acute models of inflammation and moderate activity in chronic models of inflammation.

Essential Oils from the Leaves, Stem, and Roots of Blumea lanceolaria (Roxb.) Druce in Vietnam: Determination of Chemical Composition, and In Vitro, In Vivo, and In Silico Studies on Anti-Inflammatory Activity.

Blumea lanceolaria (Roxb.) Druce, a flowering plant, is used for treating cancer and inflammatory diseases. In this study, we determined the chemical composition of the EOs extracted from the leaves (LBEO), stem (SBEO), and roots (RBEO) of B. lanceolaria and analyzed their anti-inflammation potential. Overall, 30 compounds representing 99.12%, 98.44%, and 96.89% of total EO constituents of the leaves, stem, and roots, respectively, were identified using GC-MS. ELISA, Western blotting, and qRT-PCR studies showed that LBEO, SBEO, and RBEO inhibited multiple steps in the inflammatory responses in the RAW 264.7 cell model, including NO production; TNF-α, IL-6, iNOS, and COX-2 transcription and translation; and phosphorylation of IκBα and p65 of the NF-κB pathway. In the carrageenan-induced paw edema model, all three EOs inhibited paw edema at both early and delayed phases. Molecular docking studies indicated that the main components of B. lanceolaria EOs (BEOs) targeted and inhibited major components of inflammation-related pathways, including the arachidonic acid metabolic pathway, NF-κB pathway, and MAPK pathway. We present the first study to characterize the chemical composition of BEOs and confirm their potent anti-inflammatory effects in in vitro, in vivo, and in silico analysis. These results can facilitate the development of effective anti-inflammatory drugs with limited side effects in the future.

Anti-inflammatory activity of Echinosophora koreensis nakai root extract in lipopolysaccharides-stimulated RAW 264.7 cells and carrageenan-induced mouse paw edema model

Ethnopharmacological relevanceEchinosophora koreensis Nakai is an endemic plant species distributed in a limited area within the Korean province of Gangwon, including the Yanggu-gun, Inje-gun, Cheorwon-gun, Chuncheon-si, and Hongcheon-gun counties. It is used in traditional medicine to treat various disorders, such as fever, skin diseases, diuresis, and neuralgia. Materials and methodsThis study demonstrated the effects of E. koreensis Nakai root extract (EKRE) on lipopolysaccharide (LPS)-induced inflammatory responses in vitro and in vivo. Cell viability was assessed through a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Nitric oxide (NO) production was measured using Griess reagent. Interleukin (IL)-6 and tumor necrosis factor (TNF) levels were assessed using enzyme-linked immunosorbent assays. Inducible nitric oxide synthase (iNOS), nuclear factor kappa-B (NF-κB), and mitogen-activated protein kinase (MAPK) expression were assessed using Western blot analysis. To examine the effects of EKRE in vivo, it was administered orally at doses of 50 or 200 mg/kg for 3 days in mice. Edema in the paws was induced through λ-carrageenan injection and measured hourly for up to 5 h using calipers. ResultsEKRE markedly suppressed LPS-generated NO, IL-6, and iNOS production in RAW 264.7 cells. Moreover, it suppressed the activation of the NF-κB and MAPK in LPS-stimulated cells. Furthermore, EKRE significantly inhibited carrageenan-induced edema in mouse paws. There were no significant differences in IL-6 and TNF production in paw tissue harvested from mice, but levels decreased at high EKRE concentrations (200 mg/kg). ConclusionThe results of this study provided validation for EKRE-induced inhibition of inflammatory responses in vitro and in vivo. This research suggested that EKRE is a promising treatment for inflammatory disorders.

ИЗУЧЕНИЕ ПРОТИВОВОСПАЛИТЕЛЬНОЙ АКТИВНОСТИ ВОДНОГО ЭКСТРАКТА ЛИСТЬЕВ CIRSIUM HETEROPHYLLUM (L.) HILL

The biological activity of extracts from Cirsium heterophyllum leaves is of interest. Traditional medicine and preliminary studies indicate that this raw material may have the potential for anti-inflammatory activity. The aim of this study was to evaluate the anti-inflammatory activity of an aqueous extract of Cirsium heterophyllum leaves. We studied the anti-inflammatory activity of aqueous extracts of Cirsium heterophyllum leaves harvested in Vasilyevka village, Perm district, Perm territory (kray), Russia. The extracts were standardized for the content of flavonoids in terms of cynaroside. To study the anti-inflammatory activity, we used carrageenan-induced paw edema model in outbred ICR (CD-1) mice and outbred Wistar rats. Diclofenac was used as a reference drug. The test substances were dissolved in a 2% starch solution and injected intraperitoneally to the animals. The paw volume was measured hydrometrically and photometrically. The study has shown that the aqueous extracts of Cirsium heterophyllum have a mild effect compared to diclofenac. In mice, the percentage of inhibition of edema at the 4th hour was 19% (p-value = 0.0310) and 20% (p-value = 0.0130) for photometry and hydrometry, respectively. In rats, the percentage of edema inhibition was 47% (p-value = 0.0300) at the 1st hour and 28.7% (p-value = 0.0170) at 3 hours for hydrometry; photometry did not confirm these results. The results suggest that the aqueous extract of Cirsium heterophyllum has a mild activity in comparison with diclofenac, which may be due to the mutual action of the extract substances when applied intraperitoneally, as well as the presence of polysaccharides in the aqueous extract. Further research is required to determine whether the anti-inflammatory activity of its aqueous extract might be better combined with wound healing effects when applied topically.

Chemical Composition and Antioxidant, Antimicrobial, and Anti-Inflammatory Properties of Origanum compactum Benth Essential Oils from Two Regions: In Vitro and In Vivo Evidence and In Silico Molecular Investigations.

The purposes of this investigatory study were to determine the chemical composition of the essential oils (EOs) of Origanum compactum from two Moroccan regions (Boulemane and Taounate), as well as the evaluation of their biological effects. Determining EOs' chemical composition was performed by a gas chromatography-mass spectrophotometer (GC-MS). The antioxidant activity of EOs was evaluated using free radical scavenging ability (DPPH method), fluorescence recovery after photobleaching (FRAP), and lipid peroxidation inhibition assays. The anti-inflammatory effect was assessed in vitro using the 5-lipoxygenase (5-LOX) inhibition test and in vivo using the carrageenan-induced paw edema model. Finally, the antibacterial effect was evaluated against several strains using the disk-diffusion assay and the micro-dilution method. The chemical constituent of O. compactum EO (OCEO) from the Boulemane zone is dominated by carvacrol (45.80%), thymol (18.86%), and α-pinene (13.43%). However, OCEO from the Taounate zone is rich in 3-carene (19.56%), thymol (12.98%), and o-cymene (11.16%). OCEO from Taounate showed higher antioxidant activity than EO from Boulemane. Nevertheless, EO from Boulemane considerably inhibited 5-LOX (IC50 = 0.68 ± 0.02 µg/mL) compared to EO from Taounate (IC50 = 1.33 ± 0.01 µg/mL). A similar result was obtained for tyrosinase inhibition with Boulemane EO and Taounate EO, which gave IC50s of 27.51 ± 0.03 μg/mL and 41.83 ± 0.01 μg/mL, respectively. The in vivo anti-inflammatory test showed promising effects; both EOs inhibit and reduce inflammation in mice. For antibacterial activity, both EOs were found to be significantly active against all strains tested in the disk-diffusion test, but O. compactum EO from the Boulemane region showed the highest activity. Minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) for O. compactum EO from the Boulemane region ranged from 0.06 to 0.25% (v/v) and from 0.15 to 0.21% (v/v) for O. compactum from the Taounate region. The MBC/MIC index revealed that both EOs exhibited remarkable bactericidal effects.