Protein Denaturation Assay Research Articles

In vitro Evaluation of Anti-Inflammatory Activity of Symplocos racemosa Using Protein Denaturation Assay

Introduction: Recently there is considerable awareness and interest in the field of herbal medicine due to its natural origin and lesser side effects compared to Allopathy. Selected herbal plants like Symplocos racemosa, commonly known as lodhra, are found mainly in plains and lower hills of Bengal. The word ‘Lodhra’ means ‘Propitious’. Symplocos racemosa is an important Indian traditional drug used in many Ayurvedic and herbal formulations for treatment of liver as well as uterine disorders and leucorrhea. Ethnobotanical Literature indicates use of Symplocos racemosa in treatment of eye disease, skin disease, ear disorders, liver and bowel complaints, tumours,uterine disorders, spongy and bleeding gums, asthma, fever, snakebite, gonorrhoea and arthritis.
 Aim: To analyse the anti-inflammatory activity of Symplocos racemosa using protein denaturation assay.
 Materials and Methods: 2 g of Lodhra bark powder is mixed with 100 ml distilled water & boiled for 20 min at 50°C. The extract is filtered using whatman filter paper & concentrated to 10 ml.1 ml each of Bovine serum albumin is added to various fixations of plant extract (10μL - 50 μL) and the anti - inflammatory activity was evaluated by analysing the percentage inhibition.
 Results: From this study, it is evident that Lodhra has significant anti-inflammatory activity. At 50μℓ concentration, the plant extract shower higher anti- inflammatory activity of 76%.
 Conclusion: Symplocos racemosa extract has proved to exhibit effective anti- inflammatory activity. Further studies have to be carried to analyse the other properties of this herb, which can be incorporated successfully in the pharmaceutical industry.

Pharmacological Activity of Trachyspermum ammi L. Seeds Essential Oil Grown from Northeast India

The present work has been aimed to investigate the chemical composition of the essential oil of Trachyspermum ammi L., seeds by gas chromatography/mass spectrometry method (GC/MS), antioxidant activity by DPPH, ABTS radical scavenging, reducing power assay; anti-inflammatory investigation using protein denaturation assay and anti-diabetic activity using the α-amylase inhibitory assay. The essential oil was further evaluated for antibacterial activity against various gram-positive bacteria Bacillus cereus, Bacillus subtilis, Staphylococcus aureus, Micrococcus luteus and Streptococcus mutans and gram-negative bacteria Salmonella typhimurium and Salmonella enterica and compared with the standard Ciprofloxacin. Again, the antifungal activity was investigated against Fusarium keratoplasticum, Aspergillus niger, Saccharomyces cereviaceae, Candida albicans and Aspergillus fumigatus and compared with the standard Fluconazole. The GC/MS analysis of Ajwain essential oil revealed the presence of thymol (50.43%) as the major compound followed by other minor and trace compounds. Ajwain essential oil showed better antioxidant activity than that of standard as revealed by DPPH radical scavenging assay with Ajwain essential oil IC50 3.09 μL/mL which was better performing than ascorbic acid with IC50 8.87 μL/mL. The IC50 value of 21.38 μL/mL was shown by Ajwain essential oil followed by ascorbic acid 79.13 μL/mL in ABTS assay. The in-vitro anti-inflammatory assay revealed Ajwain essential oil as a good candidate for strong natural inflammation inhibitory drug formulation with an IC50 value of 93.12 μL/mL in albumin denaturation assay. Significant α-amylase inhibitory activity was also reported with an IC50 = 22.02 μL/mL better than standard acarbose IC50 = 22.46 μL/mL. Weak antimicrobial activity was reported against all the thirteen tested bacterial and fungal strains. From all the aforementioned results it can be concluded that the presence of the polyphenolic compound thymol (50.43%) resulted in strong antioxidant, anti-inflammatory and anti-diabetic activities of Ajwain essential oil, which resembles the use of thymol in the future as natural, cost-effective and potential antioxidant, inflammation inhibitory and anti-diabetic precursor for drug formulation.

Novel Anticancer Dimeric Naphthoquinones from Diospyros lotus having Anti- Tumor, Anti-Inflammatory and Multidrug Resistance Reversal Potential: In Vitro, In Vivo and In Silico Evidence.

Cancer being a genetically heterogeneous and complex disease and the available therapies are not very effective, rendering them the predominant cause of mortality across the world. The discovery of new anticancer drugs with higher efficacy and milder side effects is a great challenge for health professionals. The current study focused on the anticancer potential of two known dimeric napthoquiones, diospyrin (1) and 8-hydroxydiospyrin (2) isolated from the roots of Diospyros lotus. In vitro Epstein-Barr-Virus (EVA) an early antigen activation assay was used to evaluate the antitumor potential of tested compounds followed by a two-stage carcinogenesis assay on mouse skin for anti-carcinogenic effect. Compounds were also assessed for their multidrug resistance reversal potential. The in vitro heatinduced protein denaturation assay was used for the anti-inflammatory effect of the tested compounds. Both compounds evoked marked cytotoxic activity with IC50 of 47.40 and 36.91 ppm, respectively. In Epstein-Barr-Virus (EVA) early antigen activation assay compounds 1 and 2 showed IC50 values of 426 ppm and 412 ppm, respectively. The tested compounds showed 60% survival rate of the lymphoblastoid Raji cells at a concentration of 1000 (mol / ratio 32 pmol TPA). In a two-stage carcinogenesis assay on mouse skin, both compounds significantly delayed the formation of papillomas on mouse skin. Compound 1 showed 50% effect at 14th week, whereas compound 2 exerted the same effect at 13th week, while both provoked 100% effect at 20th week. Both compounds significantly attenuated thermal-induced protein denaturation with EC50 values of 298 and 264 μg/mL, respectively. The dimeric napthoquiones were evaluated for their effects on the reversion of Multidrug-Resistant (MDR) cell lines mediated by P-glycoprotein using rhodamine 123 dye-based exclusion screening test on human mdr1 gene transfected thymic lymphoma L5178 cell line. The compounds 1 and 2 exhibited promising MDR reversal effect in a dose-dependent manner against mouse T-lymphoma cell line. Docking results also showed that both compounds have good docking statistics as compared with standard. Both the compounds demonstrated marked anti-tumor, anti-carcinogenic, and MDR reversal effects with significant attenuation of thermal-induced denaturation of the protein. These compounds may explain the traditional uses of D. lotus which might be effective anticancer agents.

Anti-inflammatory activity of endophytic bacterial isolates from Emilia sonchifolia (Linn.) DC.

Ethnopharmacological relevanceIn the traditional medicine system, plants have been utilized as a rich source of anti-microbial, anti-inflammatory, anti-cancer, anti-viral and anti-oxidant compounds. The biological properties of plant-based drugs depend on their interaction with endophytes which persist as an important provider of bioactive secondary metabolites. Bacterial endophytes secrete anti-inflammatory molecules whose activity can be the base for the anti-inflammatory property of the plant.Aim of the study: During the screening of endophytes from Emilia sonchifolia, we isolated six different bacteria whose potential as the sources of anti-inflamamtory compounds have been aimed at in this study. Materials and methodsAnti-inflammatory activity of the ethyl acetate extract of endophytes was studied by both in vitro and in vivo analyses. In vitro study was done using protein denaturation, COX, LOX, iNOS, myeloperoxidase and nitric oxide assays and in vivo analysis was carried out by carrageenan-induced and formalin-induced paw oedema tests. The expression level of anti-inflammatory genes such as COX-2 and NfKb was confirmed by real time PCR. ResultsWe confirmed anti-inflammatory activity of the ethyl acetate extract of bacterial endophytes of E sonchifolia by both in vitro and in vivo experiments. Carrageenan- and formalin-induced inflammations in mice were effectively reduced by the administration of the bacterial extract. Among the isolates, strain ES1effectively reduced inflammation. Gene expression studies confirmed reduction in the expression of COX-2 and NfKb genes in the presence of ES1 extract. ConclusionThe present investigation demonstrated the anti-inflammatory property of the isolated bacterial endophyte ES1 (Bacillus subtilis strain-MG 692780) and thus justifies the possible role of endophytes in contributing anti-inflammatory property to E sonchifolia which is ethno-botanically important as a source of anti-inflammatory drug.

HPTLC analysis and anti-inflammatory activity of Plectranthus amboinicus (Lour.) leaves

Plectranthus amboinicus (Lour.) Spreng. are generally used in Chinese folk medicine for treating fever, cough, mumps, sore throats, and mosquito bite. The current study was therefore carried out to provide vital pharmacological details about the leaf. This study investigated the anti-inflammatory properties of ethanolic extract from Plectranthus amboinicus. The leaves of Plectranthus amboinicus were rinsed, air dried and then powdered using machine. The ethanol extract of the plant was used for phytochemical analysis to identify the constituents present in the plants. HPTLC analysis was carried out to investigate its secondary metabolite profile by using the solvent mixture. The anti-inflammatory effect was analysed by inhibition of protein denaturation assay. The hypotonicity induced haemolysis was observed from 100µg/ml to 500µg/ml of extract concentration. The results proved that the ethanolic leaf extract of P.amboinicus has antiinflammatory activity. The presence of flavonoid, quercetin in plant extract was confirmed by HPTLC analysis and these flavonoids may be responsible for the antiinflammatory activity. Further studies are needed to the development of potential drug that may be used for various pharmacological activities.

Investigation of anti-inflammatory and anti-arthritic potentials of Terminalia catappa bark using in vitro assays and carrageenan-induced inflammation, complete Freund's adjuvant induced arthritis model in rats

Terminalia catappa Linn (T. catappa) belongs to the family Combretaceae, locally known as Bengal Almond. The plant is known to have many phytochemicals such as carbohydrates, phytosterols, tannins, polyphenols, flavonoids, and saponins. It is widely used in the treatment of various diseases such as dermatitis, leprosy, and scabies and internally for colic, headache and hepatitis and also, the plant possesses anticancer, antioxidant, antidiabetic, hepatoprotective and extracts have been shown to modify reproductive behavior in rats. The main objective of this study is to assess the pharmacological potentials of T. catappa bark in the therapeutic areas of inflammation and arthritis. T. catappa bark alcoholic extract (TCE) and water extract (TCW) were prepared and evaluated for its possible anti-inflammatory activity by in vitro protein denaturation assays and membrane stabilization assay. Further, the anti-inflammatory and anti-arthritic activity of bark extracts were evaluated by in vivo models such as carrageenan-induced paw edema, air-pouch model and Complete Freund's adjuvant (CFA) induced arthritis model in rats. Total phenolic content (TPC) was found to be 287 and 175 mg/g for TCE and TCW. Total flavonoid content (TFC) was found to be 10.2 and 61.7 mg/Kg for TCE and TCW. The concentration required for the 50 % inhibition of the protein denaturation (IC 50) was found to be lower for TCE (36.2 µg/mL) in egg albumin denaturation assay, whereas in the membrane stabilization assay method TCW exhibited a lower IC50 value in comparison with diclofenac. Both the extracts showed activity in the in vitro models of inflammation and were also found to be effective in reducing the paw edema in the carrageenan-induced inflammation model and inhibition of cellular infiltration and Myeloperoxidase activity in air pouch fluid, suggesting its anti-inflammatory activity. In CFA-induced arthritis model, the extracts exhibited anti-arthritic potential by improving the altered blood parameters, arthritic scores, radio-graphical, and histopathological changes. The results of the study suggested that the bark of T. catappa possessed anti-inflammatory and anti-arthritic activities.

Chemical composition of Callistemon citrinus (Curtis) Skeels aerial part essential oil and its pharmacological applications, neurodegenerative inhibitory, and genotoxic efficiencies.

Callistemon citrinus aerial part essential oil (CCEO) was analyzed for chemical compositions using GC/MS. Pharmacological activities such as neurodegenerative inhibitory, antioxidant, anti-inflammatory, antimicrobial, and acetylcholinesterase activities were evaluated using the standard methodologies. Genotoxicity was investigated using Allium cepa assay. GC/MS analysis identified 27 compounds; eucalyptol (55.40%) was the major component. Radical scavenging activity showed IC50 value of 16.71μg/mL. Protein denaturation assay showed IC50 value of 21.19μg/mL and 19.53μg/mL in protease inhibitor activity. MIC assay revealed antimicrobial potential of CCEO against microbial strains B. cereus at 2.00mg/mL, S. typhimurium at 4.50mg/mL, S. mutans at 2.50mg/mL, C. albicans at 4.00mg/mL, and S. cerevisiae at 4.50mg/mL concentrations. Mitotic index value (MI) of CCEO showed negligible genotoxicity with MI 17.25% close to distilled water 18.22%. Acetylcholinesterase strong inhibitory activity of CCEO was observed from IC50 =6.335μg/mL. CCEO could be a cheap and easy source for the extraction of the pure compound eucalyptol and possess various biological activities which increase its pharmacological value as well as its applicability in the field of flavor and fragrance industries. PRACTICAL APPLICATIONS: Eucalyptol is the major component of many pharmaceutical and food/flavor industries. The present investigation provides a new source for the isolation of pure compound eucalyptol in cost-effective way. Additionally, the essential oil could also be used for the pharmaceutical formulation of antioxidant, inflammation inhibitory, and neurodegenerative inhibitory drug preparation under the safety range concentration.

4-Aminoantipyrine Analogs as Anti-inflammatory and Antioxidant agents: Synthesis, Biological Evaluation and Molecular Docking Studies

Objectives: A novel series of 4-aminoantipyrine derivatives were designed and efficiently synthesized from Gewald, triphosgene and various substituted aromatic/ aliphatic/heterocyclic amines. Methods: The synthesized derivatives were characterized by IR, NMR, Mass and elemental analysis. The synthesized derivatives were evaluated for their anti-inflammatory activity using in vitro protein denaturation assay and antioxidant activity by using the 1,1-diphenyl-1-picrylhydrazyl free radical scavenging method. To establish the selectivity and safety profile of the drug, the most active compounds 4a and 4b were further screened for cytotoxicity against HeLa and MCF-7 cell lines using the sulforhodamine-B assay. The synthesized compounds were also analyzed for ADME properties and a docking study was done into the active site of oxidoreductase, cyclooxygenase-1 and cyclooxygenase-2 enzymes in an attempt to understand their binding mode using Auto Dock Vina. Results: Among the series Compounds 4a and 4b showed potent anti-inflammatory activity and antioxidant activity as compared with standard diclofenac and Ascorbic acid respectively. The results of in silico ADME Screening showed that compounds could be exploited as an oral drug candidate. The most prominent compound 4a and 4b showed no significant cytotoxic activity against HeLa and MCF-7 cell lines. The molecular docking study of most active compounds had shown good binding interactions against oxidoreductase, cyclooxygenase-1 and cyclooxygenase-2 enzymes. Conclusion: Results of in vitro anti-inflammatory, antioxidant activities and docking study showed that synthesized compounds had potential antiinflammatory and antioxidant activities and can be further optimized and developed as a lead compound.

Preliminary Investigation of the Antioxidant, Anti-Diabetic, and Anti-Inflammatory Activity of Enteromorpha intestinalis Extracts

Marine algae are a promising source of potent bioactive agents against oxidative stress, diabetes, and inflammation. However, the possible therapeutic effects of many algal metabolites have not been exploited yet. In this regard, we explored the therapeutic potential of Enteromorpha intestinalis extracts obtained from methanol, ethanol, and hexane, in contrasting oxidative stress. The total phenolic (TPC) and flavonoids (TFC) content were quantified in all extracts, with ethanol yielding the best values (about 60 and 625 mg of gallic acid and rutin equivalents per gram of extract, respectively). Their antioxidant potential was also assessed through DPPH•, hydroxyl radical, hydrogen peroxide, and superoxide anion scavenging assays, showing a concentration-dependent activity which was greater in the extracts from protic and more polar solvents. The α-amylase and α-glucosidase activities were estimated for checking the antidiabetic capacity, with IC50 values of about 3.8 µg/mL for the methanolic extract, almost as low as those obtained with acarbose (about 2.8 and 3.3 µg/mL, respectively). The same extract also showed remarkable anti-inflammatory effect, as determined by hemolysis, protein denaturation, proteinase and lipoxygenase activity assays, with respectable IC50 values (about 11, 4, 6, and 5 µg/mL, respectively), also in comparison to commercially used drugs, such as acetylsalicylic acid.

A Comparative Analysis of Bark and Leaf Essential Oil and their Chemical Composition, Antioxidant, Anti-inflammatory, Antimicrobial Activities and Genotoxicity of North East Indian Cinnamomum zeylanicum Blume

Background: Cinnamomum zeylanicum Blume ( Lauraceae) bark and leaf essential oil possesses eugenol as the major component. Both the essential oils have the potential antioxidant, anti- inflammatory, antimicrobial and negligible genotoxicity. Objective: A comparative chemical composition and biological evaluation of Cinnamomum zeylanicum Blume, bark and leaf essential oils, originated from North-East India has been investigated in the present study. Methods: Gas chromatography/mass spectroscopy analysis was used to investigate the chemical composition of the essential oil samples. DPPH free radical scavenging activity, and reducing power assay were used for antioxidant activity evaluation. Similarly, egg albumin denaturation assay, protease inhibitory activities were used for anti-inflammatory activity, while genotoxicity was evaluated by using Allium cepa assay and antimicrobial activities using disc diffusion and minimum inhibitory concentration assay. Results: The GC/MS results showed that eugenol is the major component of C. zeylanicum bark (84.48%) and leaf (48.78 %) essential oil. Significant antioxidant activity was observed for both essential oils in the DPPH free radical scavenging assay (IC50= 103.2 μg/mL, bark; IC50= 234.7 μg/mL, leaf), and reducing power assay (absorbance of 1.802 nm, 0.907 nm in 48 μg/mL for bark and leaf essential oil respectively). In-vitro anti-inflammatory activity revealed strong potential of leaf essential oil (protein denaturation assay, IC50= 0.05279 μg/mL; protease inhibitory activity IC50= 3.607 μg/mL) in comparison to bark essential oil (protein denaturation assay IC50= 0.1491, protease inhibitory activity IC50= 61.06). Allium cepa root growth test, mitotic index, and chromosomal aberration test were performed to investigate the genotoxicity of C. zeylanicum bark and leaf essential oil, which resulted in no toxic nature of both the essential oils. Antimicrobial activity against eight bacterial and fungal strains revealed good antimicrobial properties with the MIC value of Staphylococcus aureus at 5 μL/mL (bark), Aspergillus niger at 2.5 μL/mL (bark); Bacillus cereus at 5 μL/mL (leaf) and Aspergillus niger at 2.5 μL/mL (leaf). Conclusion: Collectively, these findings indicated the strong biological potential of both essential oils, although bark essential possesses better biological activities than the leaf essential oil and could be used in the food and beverage industries.

Effect of different polarity solvents on the anti‐inflammatory activity of Symplocos cochinchinensis leaves and correlation with total polyphenol content

AbstractSymplocos cochinchinensis leaves known as Vietnamese traditional herbal medicines which have been utilized anciently to support the medication of infirmities. The study aimed to explore the correlation of total polyphenol content (TPC) with anti‐inflammatory activity of S. cochinchinensis leaves. S. cochinchinensis leaf extracts (ethanol extract, chloroform, ethyl acetate, n‐butanol, aqueous fractions) was determined the TPC by Folin‐Ciocalteu reagent, anti‐inflammatory proficiency by in vitro protein denaturation and membrane lysis assays. The correlation between TPC and anti‐inflammatory activities was analyzed in silico by Pearson's method. Results illustrated that the TPC was arranged in sort of descending order as ethyl acetate fraction > n‐butanol fraction > ethanol fraction > aqueous fraction > chloroform fraction. S. cochinchinensis leaf extracts exhibited anti‐inflammatory effect via their protein denaturation inhibition capacity as well as hemolysis by heat and hypotonicity, which was better than the standard drug diclofenac sodium. There was a significantly negative correlation between TPC in S. cochinchinensis leaf extracts with their IC50 inhibition of protein denaturation, hemolysis by heat and hypotonicity with Pearson's correlation coefficient (r) of ‐0.6302, ‐0.7241, and ‐0.9101, respectively. It was suggested that S. cochinchinensis leaves have great potential for anti‐inflammatory activity during the high total polyphenol content. Hence, polyphenol compounds of S. cochinchinensis leaves should be considered since they can have good effects when in pure form.

Rosuvastatin Attenuates Rheumatoid Arthritis-Associated Manifestations via Modulation of the Pro- and Anti-inflammatory Cytokine Network: A Combination of In Vitro and In Vivo Studies.

The current investigation employed rosuvastatin for evaluation as an antiarthritic agent by in vitro and in vivo studies. In vitro studies comprised egg albumin and bovine serum albumin protein denaturation assays along with membrane stabilization assays, while in vivo studies comprised formaldehyde and complete Freund’s adjuvant (CFA)-provoked arthritis. The antioxidant potential was estimated via DPPH free radical scavenging and ferric reducing assays. Rosuvastatin significantly inhibited heat-provoked protein denaturation of egg albumin and bovine serum in a concentration-dependent way with the highest inhibition of 1225 ± 9.83 and 82.80 ± 4.03 at 6400 μg/mL. The percentage protection of the RBC membrane from hypotonicity-prompted lysis was found to be 80.67 ± 2.7. Rosuvastatin promisingly subdued formaldehyde-provoked arthritis, with maximum reduction (65.47%) of the paw volume being observed at a dose of 40 mg/kg. Rosuvastatin also significantly (p < 0.001) attenuated arthritis induced by CFA injection by reducing the paw volume and arthritic index. The reduction in the body weight due to CFA injection was also preserved by rosuvastatin treatment. Hematological and biochemical changes due to arthritis induction by CFA injection were also maintained near normal values by rosuvastatin. The histopathological and radiographic investigation also revealed the protective effect of rosuvastatin on preventing structural changes. Gene expression of IL-1β, TNF-α, and IL-6 was reduced, while IL-4 and IL-10 levels were elevated by rosuvastatin in comparison to those for the disease control group. Concentration-dependent antioxidant potential was shown by rosuvastatin. Thus, rosuvastatin possesses a notable antiarthritic potential as evidenced via in vitro and in vivo studies.

In-Vivo and In-Vitro Anti-Inflammatory Activities of the Aqueous Extract of Di-Herbal Formulation (Euphorbia hirta and lactuca virosa)

Euphorbia hirta and Lactuca virosa are medicinal plants that have been used in the cure and treatment of various diseases and for health care. This study aims at evaluating the anti-inflammatory activities of the aqueous extracts of Euphorbia hirta and Lactuca virosa plants. Albumin and histamine induced inflammation in mice and xylene induced ear edema were used for the in-vivo anti-inflammatory studies. Erythrocyte membrane stabilization and inhibition of protein denaturation assays were used for the in-vitro anti-inflammatory studies. Combined doses of 100 mg/kg Euphorbia hirta and 50 mg/kg Lactuca virosa, 100 mg/kg Euphorbia hirta and 100 mg/kg Lactuca virosa, 100 mg/kg Lactuca virosa only and 10 mg/kg diclofenac significantly reduced inflamed paw in mice (P&lt;0.05) compared to control in albumin and histamine induced inflammatory test. Combined doses of 50 mg/kg Euphorbia hirta and 100 mg/kg Lactuca virosa,100 mg/kg Euphorbia hirta, 100mg/kg Lactuca virosa significantly reduced xylene induced inflammation (P&lt;0.001) compared to control. The extracts at 1 mg/ml, 2 mg/ml and 3 mg/ml significantly inhibited protein denaturation (P&lt;0.001) and heat induced hemolysis of erythrocytes (P&lt;0.0001). The plant extract of Euphorbia hirta and lactuca virosa possesses in-vivo and in-vivo anti-inflammatory effects.&#x0D; Keywords: Euphorbia hirta, Lactuca virosa, anti-inflammatory, protein denaturation

Syntheses, Characterization and Biological Activity of Novel Thiazoylazo Dye and Its Coordination Compounds

The present work describes the synthesis of a novel heterocyclic azo dye by general diazonization of 2-amino-4,5-dimethylthiazole followed by the diazo- coupling of the resulting diazonium ion with 5-methyl-2-(propan-2-yl)phenol to obtain ligand L. This was characterized using Fourier-transformed infrared and electronic spectrophotometry. Ligand L was further coordinated with five metal ions, M:L, 1:2 [M = Cu(II), Mn(II), Zn(II), Ni(II) and Co(II)]. The coordination compounds obtained were characterized by electronic, IR spectrophotometry, magnetic susceptibility and percentage metal analyses. The results obtained suggested that a thiazoylazo dye was obtained as ligand L. It was proposed that two molecules of the solvent coordinated to the metal ion in addition with the ligands to give an octahedral geometry for copper(II), manganese(II) and nickel(II) complexes. On the other hand, square planar geometry was suggested for zinc(II) and cobalt(II) complexes. The anti-infla- mmatory activity of the ligand and coordination compounds was evaluated using four in vitro-based assays viz: xanthine oxidase and lipoxygenase inhibition assay, membrane stability and protein denaturation assay. The synthesized compounds generally exhibited good anti-inflammatory activity in all the assays carried out. However, the reference standards, in this instance, were more effective in the case of xanthine oxidase, lipoxygenase and protein denaturation inhibitory assays. For the membrane stability study, the coordination compounds and ligand L elicited more potent anti-inflammatory activity than the standard drug.

Methanolic Extract of Distemonanthus benthamianus (Caesalpiniaceae) Stem Bark Suppresses Ethanol/Indomethacin-Induced Chronic Gastric Injury in Rats.

Distemonanthus benthamianus (Caesalpiniaceae) is a plant from the Cameroon pharmacopoeia very widely used in the treatment of many pathologies among which are gastrointestinal disorders. The main purpose of this study was to assess the healing properties of gastric ulcer from the methanolic extract of Distemonanthus benthamianus and its mechanisms of action. The healing properties of gastric ulcers (chronic ulcer model induced by ethanol and indomethacin) were evaluated in vivo in adult male rats, while the mechanisms of action were evaluated in vitro by anti-inflammatory assay (protein denaturation, cyclooxygenase, and lipoxygenase assays) and immunomodulatory assay (ROS production (using technical chemiluminescence), cytokine (TNF-α, IL-1β, IL-6) production (using ELISA), proliferation of T cells (using liquid scintillation counter), and cytotoxicity (using MTT assay)). The methanolic extract of Distemonanthus benthamianus inhibited protein denaturation (75.63%) and the activities of cyclooxygenase (78.92%) and 5-lipoxygenase (81.54%). The extract also significantly (p < 0.001) inhibited intracellular and extracellular ROS production and T cell proliferation and reduced significantly (p < 0.01, p < 0.001) TNF-α, IL-1β, IL-6, and PGE2 production. At all doses (125, 250, and 500 mg/kg), the extract significantly reduces the ulceration index and the area of ulceration and significantly increases the mass of gastric mucus. In addition, the extract significantly decreases the level of MDA, significantly increases the activities of catalase and glutathione, and then improves the hematological parameters in sick animals. Histological micrographs show that in the presence of the extract, there is advanced reepithelialization with recovery of the ulcerated epithelium. Thus, the extract of Distemonanthus benthamianus has healing properties against gastric ulcers which are associated with its anti-inflammatory, immunomodulatory, and antioxidant effects.

Synthesis, 3D-QSAR, and Molecular Modeling Studies of Triazole Bearing Compounds as a Promising Scaffold for Cyclooxygenase-2 Inhibition.

Targeting of cyclooxygenase-2 (COX-2) has emerged as a powerful tool for therapeutic intervention because the overexpression of this enzyme is synonymous with inflammation, cancer, and neurodegenerative diseases. Herein, a new series of 1,2,4-triazole Schiff bases scaffold with aryl and heteroaryl systems 9a–12d were designed, synthesized, structurally elucidated, and biologically evaluated as a potent COX-2 blocker. The rationale beyond the current study is to increase the molecule bulkiness allowing a selective binding to the unique hydrophobic pocket of COX-2. Among the triazole–thiazole hybrids, the one with the para-methoxy moiety linked to a phenyl ring 12d showed the highest In vitro selectivity by COX-2 inhibition assay (IC50 of 0.04 μM) and in situ anti-inflammatory activity when evaluated using the protein denaturation assay (IC50 of 0.88 μM) in comparison with commercially available selective COX-2 inhibitor, Celecoxib (IC50 of 0.05 μM). Towards the COX-2 selectivity, ligand-based three dimensional quantitative structures activity relationship (3D-QSAR) employing atomic-based and field-based approaches were performed and resulted in the necessity of triazole and thiazole/oxazole scaffolds for COX-2 blocking. Furthermore, the molecular modeling study indicated a high selectivity and promising affinity of our prepared compounds to COX-2, especially the hydrophobic pocket and the mouth of the active site holding hydrogen-bonding, hydrophobic, and electrostatic interactions. In Silico absorption, delivery, metabolism, and excretion (ADME) predictions showed that all the pharmacokinetic and physicochemical features are within the appropriate range for human use.

North-East Indian Chromolaena odorata (L. King Robinson) Aerial Part Essential Oil Chemical Composition, Pharmacological Activities - Neurodegenerative Inhibitory and Toxicity Study

Current study was formulated to analyze the chemical composition, pharmacological activities, and toxicology study of Chromolaena odorata aerial part essential oil (COEO) grown in North East India. Gas Chromatography/Mass Spectroscopy was used for chemical composition analysis. The antioxidative potential was tested using DPPH and reducing power assays. Protein denaturation and protease inhibitor assays for anti-inflammatory activity and Allium cepa assay were used to test the genotoxicity of the essential oil. Standard disc diffusion and minimum inhibitory assays (MIC) were employed to test the antimicrobial efficacy of COEO and Acetylcholinesterase (AChE) inhibitory assay was used for anticholinesterase activity. GC/MS analysis revealed Geijerene (26.34 %), α-Copaene (17.87 %), Caryophyllene (11.14 %), 3-Carene (10.63 %), Cadinene (7.98 %), Cyclohexane, 1-Methylene-4-(1-methylethenyl)-(3.37 %), Elemol (2.89 %), τ-Muurolol (2.77 %), as major components. Standard DPPH, protein denaturation, and protease inhibitor assays revealed (IC50= 15.88 μg/mL, 19.35 μg/mL, and 27.28 μg/mL respectively) strong antioxidant, anti-inflammatory potential of COEO. Allium cepa assay showed negative genotoxicity with (mitotic index) MI=16.38 %, which is close to distilled water MI=19.20 %. While disc diffusion assay (zone of inhibition for B.cereus 16.56 ± 0.002 mm; S. aureus 14.20 ± 0.057 mm; S. cerevisiae 10.67 ± 0.015 mm; A. fumigatus 10.15 ± 0.05 mm) and MIC (B. subtilis, S. aureus 2.50 μL/mL), showed the antimicrobial potential of COEO. Strong AChE inhibitory activity was observed with an IC50 value of 61.98 μg/mL. From all the results obtained it is clear that COEO could be used for future research, development, and in preparation of cost-effective pharmacologically important herbal drugs after proper validations.

Ethnomedicinal documentation and anti-inflammatory effects of n-butanol extract and of four compounds isolated from the stems of Pituranthos scoparius: An in vitro and in vivo investigation

Ethnopharmacological relevancePituranthos scoparius is a medicinal plant belonging to the Apiaceae family. It thrives in North Africa, and is widely distributed in the high plateau of most parts of the Sahara in Algeria. This plant is widely used in the Algerian traditional medicine to treat numerous infectious diseases, dermatoses, nervous breakdowns, digestive disorders, and calm abdominal pain. Aim of the studyThe aim of the present work was to examine the ethnomedicinal uses of Pituranthos scoparius in Setif region, Algeria, isolate and identify the chemical constituents of the n-butanol stem extract of P. scoparius (BEPS), and to determine the toxicity and anti-inflammatory effects of these compounds in addition to the extract. Materials and methodsThe anti-inflammatory effects of BEPS and the four compounds isolated from the extract were evaluated using the in vitro protein denaturation assay, whereas the topical anti-inflammatory activity was assessed using the croton oil-induced ear edema in mice. Toxicity was determined based on assessment of in vitro cytotoxicity using hemolytic activity against human red blood cells (RBCs). ResultsFour compounds, identified as the rare isorhamnetin-3-O-β-apiofuranosyl (1 → 2)-β glucopyranoside (2), in addition to three known compounds, namely isorhamnetin-3-O-β-glucoside (1), D-mannitol (3), and isorhamnetin-3-O-β-glucopyranosyl-(1 → 6)-β-glucopyranoside (4) were isolated from BEPS. These compounds were characterized by means of NMR and high-resolution mass spectral (HRMS) data. These four compounds were isolated for the first time from this traditional Algerian medicinal plant. Screening of the extract indicated the presence of alkaloids, polyphenols, flavonoids, free quinones, coumarins and tannins. Topical anti-inflammatory effect showed that the four isolated compounds, as well as BEPS, exhibit a significant (p < 0.05) dose-dependent (0.5 and 1 mg/ear) anti-inflammatory effect. At a dose of 1 mg/ear, compounds 1, 2, and 4, exhibited remarkable anti-inflammatory effect with a percentage inhibition of 85.50 ± 2.78, 79.78 ± 4.68, and 75.78 ± 2.98%, respectively. Results from in vitro cytotoxicity showed that the % lysis of the extract, along with isolated compounds was found to be virtually nontoxic. ConclusionsThese results suggest that BEPS and isolated compounds are safe, nontoxic, and exert remarkable anti-inflammatory effects, and can be new sources of natural anti-inflammatory agents.

Antioxidant, Anti-inflammatory and Biosorption Properties of Starch Nanocrystals In Vitro Study: Cytotoxic and Phytotoxic Evaluation

This study aimed to investigate the antioxidant, anti-inflammatory and biosorption properties of starch nanocrystals (SNC). The characterization of synthesized SNC was done using various analytical techniques like microscopic and spectroscopic analysis. The antioxidant property was determined using DPPH (2,2-diphenyl-1-picrylhydrazyl) assay and metal ion chelating assay. SNC showed the highest scavenging activity of 70.03 ± 0.74% at 100 µg/mL concentration. Protein denaturation assay and proteinase inhibitory assay depicted the anti-inflammatory property of SNC. The results revealed that the maximum inhibition activity was found at 100 µg/mL with 72.71% inhibition. The maximum removal efficiency was found to be 83.42% at pH 2.0 with 0.15 g biosorbent. As the pH increases, biosorption capacity of SNC were reduced from 8.17 to 6.30 mg/g and the efficiency of the dye removal was decreased from 80.95 to 36.01%. The shape of synthesized SNC was spherical nanoplatelets and it shows agglomeration. The Langmuir isotherm model is best suited for the biosorption experiments with the R2 value of 0.986. SNC were subjected to cytotoxic and phytotoxic evaluation. Cell viability and phytotoxic assays proves the non-toxic nature of the SNC.

Anti-inflammatory potential of novel hexapeptide derived from Meretrix meretrix foot and its functional properties.

The study aimed to identify bioactive peptide from Meretrix meretrix Linnaeus foot (MMF) and examine its potential of suppressing inflammation. In brief, the anti-inflammatory activity was identified by erythrocyte membrane protection and protein denaturation assay from MMF peptic 9th-h hydrolysate and was separated with three molecular weight cut-off units. The obtained four fractions were testified for activity and the fraction (10-3kDa) with maximum activity was purified using gel permeation chromatography. Finally, the peptide sequence was identified as Asn-Pro-Ala-Gln-Asp-Cys (647.559Da) by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The hexapeptide was characterised for functional properties at different pH range. The non-toxic hexapeptide was able to reduce the cyclooxygenase (COX)-2 activation, pro-inflammatory cytokines and nitric oxide (NO) production significantly in RAW264.7 macrophage cells. The current results propose that the hexapeptide derived from MMF protein can act as an effective anti-inflammatory against pro-inflammatory cytokines, COX-2 and NO. Moreover, it could be used as an effective alternative source for drugs in pharma and also as an ingredient in food industries.