Heat-induced Albumin Denaturation Research Articles

Mentha rotundifolia (L.) Huds. and Salvia officinalis L. hydrosols mitigate aging related comorbidities in rats.

Aging is often linked to oxidative stress, where the body experiences increased damage from free radicals. Plants are rich sources of antioxidants, playing a role in slowing down aging and supporting the proper functioning and longevity of cells. Our study focuses on exploring the impact of Mentha rotundifolia (MR) and Salvia officinalis (SO) hydrosols on aging-related comorbidities. The chemical composition of MR and SO hydrosols was analyzed by gas chromatography coupled to mass spectrometry. 2,2-Diphenyl 1-picrylhydrazyl and 2,20-azino-bis 3-ethylbenzothiazoline-6-sulfonic acid radicals scavenging assays were used to assess their in vitro antioxidant activity, and heat induced albumin denaturation test was used to evaluate their anti-inflammatory activity. Subsequently, we administered 5% of each plant hydrosol in the drinking water of 18-month-old rats for six months. We then conducted behavioral tests, including open field, dark/light box, rotarod, and Y-maze assessments, and measured biochemical parameters in plasma, liver and brain tissues. At two years old, animals treated with MR and SO hydrosols displayed fewer physical and behavioral impairments, along with well-preserved redox homeostasis in comparison with animals in the control group. These results highlighted the significance of MR and SO hydrosols in addressing various aspects of age-related comorbidities. The study suggests that these plant-derived hydrosols may have potential applications in promoting healthy aging and mitigating associated health challenges.

Copolymeric Micelles of Poly(ε-caprolactone) and Poly(methacrylic acid) as Carriers for the Oral Delivery of Resveratrol.

In this study, we report the development of a micellar system based on a poly(methacrylic acid)-b-poly(ε-caprolactone)-b-poly(methacrylic acid) triblock copolymer (PMAA16-b-PCL35-b-PMAA16) for the oral delivery of resveratrol. The micellar nanocarriers were designed to comprise a PCL core for solubilizing the poorly water-soluble drug and a hydrated PMAA corona with bioadhesive properties for providing better contact with the gastrointestinal mucosa. The micelles were first formed in an aqueous media via the solvent evaporation method and then loaded with resveratrol (72% encapsulation efficiency). Studies by transmission electron microscopy (TEM) and dynamic and electrophoretic light scattering (DLS and PALS) revealed a spherical shape, nanoscopic size (100 nm) and a negative surface charge (-30 mV) of the nanocarriers. Loading of the drug slightly decreased the hydrodynamic diameter (Dh) and increased the ƺ-potential of micelles. In vitro dissolution tests showed that 80% and 100% of resveratrol were released in 24 h in buffers with pH 1.2 and 6.8, respectively, whereas for the same time, not more than 10% of pure resveratrol was dissolved. A heat-induced albumin denaturation assay demonstrated the advantage of the aqueous micellar formulation of resveratrol, which possessed anti-inflammatory potential as high as that of the pure drug. Further, the micellar resveratrol (5 µM) exerted a strong protective effect and maintained viability of mucosa epithelial HT-29 cells in a co-cultural model, representing the production of inflammatory cytokines. For comparison, the pure resveratrol at the same concentration did not protect the damaged HT-29 cells at all. Thus, the present study revealed that the PMAA-b-PCL-b-PMAA copolymeric micelles might be considered appropriate nanocarriers for the oral delivery of resveratrol.

Novel Derivatives of Eugenol as a New Class of PPARγ Agonists in Treating Inflammation: Design, Synthesis, SAR Analysis and In Vitro Anti-Inflammatory Activity.

The main objective of this research was to develop novel compounds from readily accessed natural products especially eugenol with potential biological activity. Eugenol, the principal chemical constituent of clove (Eugenia caryophyllata) from the family Myrtaceae is renowned for its pharmacological properties, which include analgesic, antidiabetic, antioxidant, anticancer, and anti-inflammatory effects. According to reports, PPARγ regulates inflammatory reactions. The synthesized compounds were structurally analyzed using FT-IR, 1HNMR, 13CNMR, and mass spectroscopy techniques. Molecular docking was performed to analyze binding free energy and important amino acids involved in the interaction between synthesized derivatives and the target protein. The development of the structure-activity relationship is based on computational studies. Additionally, the stability of the best-docked protein-ligand complexes was assessed using molecular dynamic modeling. The in-vitro PPARγ competitive binding Lanthascreen TR-FRET assay was used to confirm the affinity of compounds to the target protein. All the synthesized derivatives were evaluated for an in vitro anti-inflammatory activity using an albumin denaturation assay and HRBC membrane stabilization at varying concentrations from 6.25 to 400 µM. In this background, with the aid of computational research, we were able to design six novel derivatives of eugenol synthesized, analyzed, and utilized TR-FRET competitive binding assay to screen them for their ability to bind PPARγ. Anti-inflammatory activity evaluation through in vitro albumin denaturation and HRBC method revealed that 1f exhibits maximum inhibition of heat-induced albumin denaturation at 50% and 85% protection against HRBC lysis at 200 and 400 µM, respectively. Overall, we found novel derivatives of eugenol that could potentially reduce inflammation by PPARγ agonism.

Sequential analysis for identification of byproduct from N-benzylation reaction: wound healing and anti-inflammatory potential of the byproduct 4-chlorobenzyl 2-((4-chlorobenzyl)amino)benzoate.

A very common reaction, N-benzylation of isatoic anhydride in the presence of sodium hydride base, produces byproducts. The yield of one of the byproducts was greater than that of the desired product; therefore, we identified the anonymous undisclosed structure of the byproduct using sequential spectroscopy methods and SC-XRD. This byproduct was found to be effective as a wound-healing and anti-inflammatory agent. The 10% formulation of byproduct and standard (nitrofurazone) showed complete wound closure with a large number of cell migrations within 16 days. Hydroxyproline contents of 5% and 10% formulations were found to be slightly increased as compared with that of the standard. The byproduct also had anti-inflammatory potential. It was effective in inhibiting COX-2, heat-induced albumin denaturation, and formalin-induced paw edema.

Therapeutic Potential of Myrrh, a Natural Resin, in Health Management through Modulation of Oxidative Stress, Inflammation, and Advanced Glycation End Products Formation Using In Vitro and In Silico Analysis

Oxidative stress, hyper-inflammatory responses, and protein glycation are the chief contributing factors in the pathogenesis of several diseases. This study aimed to explore the therapeutic role of myrrh in health management through in vitro and in silico studies. Antioxidant potential, anti-inflammatory potential, antiglycation, and advanced glycation end products (AGEs) formation inhibition activities were determined by various in vitro assays. Molecular docking was performed to predict the non-covalent binding of macromolecules (receptor) and a small molecule (ligand). Myrrh extract contained significant antioxidant activity as reflected by FRAP value (16.12 µg ascorbic acid/100 mg dry weight), the maximum percentage of DPPH scavenging (57.71%), and maximum hydrogen peroxide reducing activity (58.71%) at a concentration of 600 µg/mL. Further, the extract exhibited maximum protection from bovine serum albumin (BSA) denaturation inhibition (53.47%), anti-proteinase action (43.517%), and egg albumin denaturation inhibition (44.95%) at a dose of 600 µg/mL concentration. Myrrh is used in pharmacy as an antiseptic, anti-inflammatory, antimicrobial, antifungal, and anti-venom remedy. This study aimed to explore the antioxidant, anti-inflammatory, antiarthritic, antiglycation, and advanced glycation end products (AGEs) formation inhibition ability of methanolic myrrh extract. It was found that myrrh has good antioxidant potential due to plenty of flavonoids and polyphenolic compounds, as reflected by results of the 2,2′-diphenyl-1-picrylhydrazyl radical (DPPH) assay, FRAP (ferric reducing antioxidant power) test, and hydrogen peroxide reducing test. Furthermore, myrrh methanolic extract was found to be significantly effective against heat-induced albumin denaturation, and percent inhibition of denaturation increases with increased extract concentration. The presence of myrrh extract at a dose of 600 μg/mL decreased browning intensity (59.38%), percent aggregation index (59.88%), and percentage amyloid structure (56.13%). We used molecular docking tools to study the role of myrrh in oxidative stress (Catalase), antioxidant property (Superoxide dismutase), and antiviral property (spike protein S). The molecular docking analysis confirmed four phytoconstituents; 2,3-Furandione, Curzerene, delta-Elemene, and Furanoeudesma-1,3-Diene interact with catalase and superoxide dismutase. Curzerene and Furanoeudesma-1,3-Diene showed remarkable interaction with SARS-CoV-2 spike protein S. Our data suggest that myrrh resin extract can be used to develop a suitable alternative therapy for various diseases linked with oxidative stress, inflammation, glycation, and AGEs.

Phytochemical, Antioxidant, Anti-Inflammatory, and Thrombolytic Properties of Cleisomeria lanatum (Lindl.) Lindl. ex G. Don.

For the first time, Cleisomeria lanatum (family: Orchidaceae) has been investigated for its phytochemical, antioxidant, anti-inflammatory, and thrombolytic properties. All phytochemicals studied were identified in varying degrees during qualitative screening. In quantitative screening, a maximum of 106.02 ± 0.08 mg/g alkaloids (root), 179.67 ± 8.83 mg/g phenols (stem), 17.34 ± 0.88 mg/g flavonoids (stem), 73.67 ± 1.76 mg/g tannins (stem), and 180.04 ± 0.02 μg/mL proteins (root) were detected. Antioxidant, anti-inflammatory, and thrombolytic properties were investigated using a free-radical scavenging assay, heat-induced albumin denaturation assay, and blood clotting inhibition assay. The root extracts had the most effective antioxidant (IC50 = 67.98 μg/mL) and anti-inflammatory (IC50 = 60.86 μg/mL) properties, whereas the stem extracts had the most effective thrombolytic property (IC50 = 163.8 μg/mL). The bioactivities studied also had a significant positive relationship (r = 0.9; p < 0.05) with the amount of phenolics and tannins.

Biological Characterization and Preliminary Crystallization: A Step Towards the Use of Novel Allium sativum Protease Inhibitor as a Potential Therapeutic Drug

Background: Garlic, being a well-known medicinal plant, is the most commonly used culinary spice worldwide. Investigation of protease inhibitor isolated from garlic leads to a promising contender in pharmacognostic and pharmacological studies. Objective/Introduction: Protease Inhibitor (PI) from 'garlic' (Allium sativum) was analyzed for its biological role as an antioxidant, antimicrobial, and anti-inflammatory agent. Methods: Antioxidant activity was evaluated using ferric ion reducing antioxidant power (FRAP) and 2, 2-diphenyl-1-picrylhydrazyl (DPPH) assays. The anti-inflammatory activity was assessed using trypsin inhibitory assay and heat-induced albumin denaturation method. The antimicrobial activity was examined in broth against E. coli and B. Subtilis. The crystallization was setup using the hanging drop method. Results: ASPI showed DPPH radical scavenging with IC50 values 561±0.337 μg/ml. Also, ASPI showed the highest value of 0.699±0.009 mM at 1000 μg/ml and the lowest i.e. 0.181±0.006 mM at 100 μg/ml in FRAP assay. Ascorbic acid was taken as standard in both cases. ASPI showed IC50 values of 651±0.532 μg/ml and ~657±1.802 μg/ml, respectively. The antibacterial role of ASPI was testified, and results showed maximum inhibition against E. coli (ATCC 25922) i.e., 87.8 ±0.602% but no inhibition against B. subtilis (MTCC 736). Cuboidal shaped crystals of the ASPI were obtained in 4-6 weeks using 0.2M calcium chloride dihydrate, 0.1M sodium acetate trihydrate, 20 % isopropanol. Conclusion: ASPI has tremendous potential in the development of suitable drugs in pharmaceutical industries against diseases due to the generation of reactive oxygen species and cancer. The cuboidal crystals were obtained, which is the first study in the context of crystallization of ASPI to date.

Study of the Phytochemical Composition, the Antioxidant and the Anti-Inflammatory Effects of Two Sub-Saharan Plants: Piliostigma reticulatum and Piliostigma thonningii.

The aim of this study is to perform phytochemical screening of the leaves of Piliostigma reticulatum and Piliostigma thonningii, to determine the phenolic, flavonoids, tannins, and sugars content in their methanolic extracts, evaluate their antioxidant activity using the DPPH and the ABTS tests, and test their anti-inflammatory effect in vitro using the heat-induced albumin denaturation inhibition method. Phytochemical screening revealed the presence of polyphenols and alkaloids in the leaves of both plants. Yields of the extracts in this study ranged from 7% to 18% for P. reticulatum and 4% to 16% for P. thonningii. The phenolic content in the methanolic extract of P. reticulatum is 74.66 ± 1.76 μg GAE/mL, which is significantly higher than that of P. thonningii (56.54 ± 1.24 μg GAE/mL). Both plants showed good antioxidant activity. In fact, for the DPPH test, the IC50 value is 8.88 ± 0.11 μg/mL for P. reticulatum and 17.64 ± 0.68 μg/mL for P. thonningii. For the ABTS assay, the IC50 values of the two plants are, respectively, 9.78 ± 1.83 μg/mL and 13.47 ± 2.62 μg/ml, statistically comparable and significantly higher than the IC50 of the standard 30.76 ± 0.18 μg/ml. Leaf extracts from both plants were effective against heat-induced denaturation of albumin. The activity of P. reticulatum is indeed comparable to that of the standard with an IC50 value of 121.43 ± 1.55 μg/mL and higher than that of P. thonningii with an IC50 value of 170.15 ± 1.09 μg/mL. These results show that both plants exhibit significant antioxidant and anti-inflammatory activities. Therefore, their chemical compounds could have potential applications as antioxidant and anti-inflammatory drugs.

In-vitro antioxidant and anti-inflammatory activities of ethanol stem-bark extract of Blighia sapida K.D. Koenig

Blighia sapida (B. sapida) K.D. Koenig (Family Sapindaceae) is a branchless straight bole approximately 15 m in length. The study evaluated the antioxidant and anti-inflammatory activities of ethanol extract and fractions of B. sapida stem-bark using in vitro methods. Ethanol extract and its fractions were investigated for 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, ferric reducing antioxidant power (FRAP), total antioxidant capacity (TAC), and quantitative phenolic and flavonoid contents. Anti-inflammatory activity was evaluated using albumin denaturation and membrane stabilization assays. The extract and its fractions exhibited radical scavenging and anti-inflammatory properties. The ethyl acetate fraction possessed maximum phenolic and flavonoid contents (136.67 ± 1.55 gallic acid equivalent mg/g and 75.76 ± 4.03 quercetin equivalent mg/g, respectively). Antioxidant studies revealed that the ethyl acetate fraction displayed superior activity with an IC50 = 0.09 ± 0.03 mg/mL DPPH, and values of 146.96 ± 3.81 ascorbic acid equivalent (AAE) mg/g and 359.20 ± 4.98 AAE mg/g for FRAP and TAC, respectively. Furthermore, the anti-inflammatory activity was revealed by inhibition of heat-induced albumin denaturation and red blood cell membrane stabilization at concentrations of 200–1000 μg/mL and 50–250 μg/mL, respectively. The ethanol extract and fractions exhibited antioxidant and anti-inflammatory activities, with ethyl acetate fraction showing superior activity, which could be attributed to secondary metabolites, mainly phenolic compounds. Overall, the antioxidant and anti-inflammatory activities of B. sapida can be exploited by ethnomedicinal users.

EVALUATION OF SALVIA LANATA LEAF EXTRACT FOR ANTI-INFLAMMATORY AND ANTIOXIDANT ACTIVITY

Objective: The work is aimed to evaluate the anti-inflammatory and antioxidant activity of the ethanolic leaf extract of Salvia lanata.&#x0D; Methods: Anti-inflammatory activity of the leaf extract of S. lanata at a dose of 100 mg/kg and 200 mg/kg against the standard drug indomethacin at a dose of 10 mg/kg i.p. was evaluated by carrageenan-induced rat paw edema and protein denaturation method. Antioxidant activity was determined by 1, 1 diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging method, reducing power method, and nitric oxide scavenging assay.&#x0D; Results: S. lanata leaf extract showed highly significant dose-dependent efficacy against carrageenan-induced paw edema at a dose of 200 mg/kg and lesser effect at 100 mg/kg. It inhibited heat-induced albumin denaturation with a maximum inhibition of 79.26% at 160 μg/ml. DPPH free radical scavenging activity of extract exhibited inhibition of 25.96%–87.74% within the concentration range of 10 μg/ml–160 μg/ml, nitric oxide assay from 12.26% to 79.22% in the same concentration range. In reducing power assay with an increase in concentrations, an increase in the absorbance of the reaction mixture was observed. Antioxidant activity was compared to standard drug ascorbic acid.&#x0D; Conclusion: The leaf extract of S. lanata has potent anti-inflammatory and antioxidant activity.

EVALUATION OF ANTI-INFLAMMATORY AND ANTIDIABETIC ACTIVITY OF SIMAROUBA GLAUCA BARK EXTRACT: AN IN VITRO STUDY

Objectives: The present study was envisaged to identify the effect of anti-inflammatory and antidiabetic activity for methanolic bark extracts of Simarouba glauca. &#x0D; Methods: The present study design was to evaluate the in vitro anti-inflammatory and in vitro antidiabetic activity of S. glauca methanolic bark extract. To examine the antidiabetic activity, the samples were studied for their effect on inhibition of alpha-amylase and glucose transport across the dialysis membrane. In vitro anti-inflammatory activity was evaluated using albumin denaturation assay and membrane stabilization method.&#x0D; Results: Our current results indicate that the various bioactive constituents detected in S. glauca may be responsible for its in vitro antidiabetic and anti-inflammatory effects. The ability of plant extract on anti-inflammatory activity showed that it was effective in inhibiting heat-induced albumin denaturation with an IC50 value of test and standard was found to be 46.42 μg/ml and 24.09 μg/ml. In addition to this, heat-induced hemolysis was also performed. The IC50 values of the test and standard were found to be 43.51 μg/ml and 21.41 μg/ml, respectively. The percentage inhibition of the test sample varied from the concentration range of 75 to 100 μg/ml. The IC50 value of the test and standard was found to be 19.08 μg/ml and 9.08 μg/ml, respectively.&#x0D; Conclusion: The findings of the present study concluded that S. glauca bark has the potential to treat diabetes and a novel natural anti-inflammatory agent as a good source. Thus, S. glauca may be a potential candidate for the development of future antidiabetic and anti-inflammatory compounds. However, still further studies and standardization of the plant research may be required to develop them as medicine.

SYNTHESIS AND BIOLOGICAL EVALUATION OF NOVEL THIAZOLE-PYRAZOLE INTEGRATED CHALCONES AS ANTIOXIDANT AND ANTI-INFLAMMATORY AGENTS

Objective: The objective of the present study was to synthesize the thiazole-pyrazole integrated chalcones and their in vitro antioxidant and anti- inflammatory evaluation.&#x0D; Methods: The designed hybrid thiazole-pyrazole integrated chalcones (3a-j) were synthesized by Claisen–Schmidt reaction of substituted 1-(4-methyl-2-phenylthiazol-5-yl) ethanone and substituted pyrazole aldehyde in the presence of 10% NaOH in ethanol solvent under reflux condition. The chemical structures of synthesized compounds were confirmed by IR, 1H nuclear magnetic resonance (NMR), 13C NMR, and high- resolution mass spectra.&#x0D; Results: All the title compounds were screened for their in vitro antioxidant and anti-inflammatory activity. The screening data indicated that tested compounds showed potent antioxidant activity with moderate anti-inflammatory potential.&#x0D; Conclusion: Antioxidant screening data reveal that most of the synthesized compounds possess excellent 1,1-diphenyl-2-picrylhydrazyl and NO radical scavenging activity. Most of the compounds found to possess marked anti-inflammatory potential by effectively inhibiting the heat-induced albumin denaturation.

Anti-inflammatory activity of Punica granatum extract on oral microbes - In vitro

To study the anti-inflammatory effect of Punica granatum extract against the oral microbes. Oral diseases continue to be a major health problem worldwide. Dental caries and periodontal diseases are among the most important global oral health problems, although conditions such as oral and pharyngeal cancers and oral tissue lesions are also significant health concerns. Pomegranate extracts have been used for centuries in traditional medicine to confer health benefits in a number of inflammatory diseases, microbial infections and cancer. The anti-inflammatory activity of pomegranate extract was evaluated by protein denaturation assay, and the results were read spectrophotometrically. Denaturation of proteins is a great‐ documented cause of inflammation. As a part of the investigation on the mechanism of the anti-inflammatory activity, the ability to extract to inhibit protein denaturation was studied. It was effective in inhibiting heat induced albumin denaturation at different concentrations as shown in Table 1. Maximum inhibition, 70.12±1.12% was observed at500µg/ml. IC50 value was found to be 105.35±1.99µg/ml. Aspirin, a standard anti-inflammatory drug showed the maximum inhibition, 77.12±1.42% at the concentration of 200µg/ml. Hence it can be concluded that pomegranate extract has anti-inflammatory property and also can be used in products such as toothpaste and mouth wash etc.

Evaluation of anti-inflammatory potential of the leaves of Wendlandia heynei (Schult.) Santapau & Merchant in Sprague Dawley rat

Ethnopharmacological relevanceThe leaves of W. heynei (family: Rubiaceae) are used by the folklore in swelling, wounds and body aches. In this study anti-inflammatory potential of W. heynei leaves has been evaluated during in vitro studies and in rat. MethodsMethanol extract of the leaves of W. heynei (WHLM) and its fractions; n-hexane (WHLH), chloroform (WHLC), ethyl acetate (WHLE), n-butanol (WHLB) and residual soluble aqueous (WHLA) were screened for phytochemical analysis and several active constituents (alkaloids, flavonoids, saponins, tannins, terpenoids, β-carotene and lycopene) were also quantified. Heat induced albumin denaturation assay and in vitro cell cultures was carried out for in vitro anti-inflammatory activity, while various in vivo assays like TPA induced ear edema, croton oil induced anus edema, formalin and carrageenan-induced hind paw edema was investigated in Sprague-Dawley rats. Alterations on levels of tumor necrosis factor (TNF-α), Interleukin-1β (IL-1β), IL-6 and prostaglandins (PGE2) induced with WHLE was studied in serum after carrageenan induced paw edema in rat. Meanwhile, the dose dependent WHLE inhibition of NFκB pathway via regulation of the phosphorylation of IKKs, IκBα, and p65 subunit was studied in LPS-induced rat peritoneal macrophages. On account of marked anti-inflammatory activity of WHLE its bioactive components were analyzed by HPLC-DAD analysis. ResultsThe phytochemical analysis yielded alkaloids, saponins, tannins, coumarins, glycosides, quinones and vitamin C in WHLM and in all fractions. Fraction (WHLE) was enriched with alkaloids (20.20 ± 2.5%), flavonoids (25.26 ± 2.11%) and tannins (307.2 ± 2.03 mg of GAE/g of extract), while terpenoids (21.60 ± 1.65%) were the major constituents of WHLH. Ethyl acetate fraction convincingly protected heat induced albumin denaturation. WHLE exhibited highest edema inhibition in models of TPA-induced ear edema (74.51 ± 2.05) and croton oil-induced anal edema (75.38 ± 2.83). The pretreatment with WHLE significantly (p < 0.05) reduced the paw edema with formalin (78.99 ± 2.26%) assessed after 6 h and in carrageenan (75.71 ± 4.46%) was detected after 4 h. Level of anti-inflammatory markers; IL-1β, IL-6, TNF-α and PGE2 in carrageenan induced paw edema in serum of rat was significantly (p < 0.001) decreased with WHLE pretreatment to rat. WHLE significantly inhibited the NFκB by reducing the phosphorylation of IKKs, IκBα, and p65 subunit in LPS-induced inflammation in rat peritoneal macrophages. HPLC-DAD analysis of WHLE indicated the presence of rutin, gallic acid, catechin, caffeic acid and myricetin. ConclusionsIt is concluded that WHLM fractions have marked anti-inflammatory activity and this study endorsed the folklore use of W. heynei leaves for swelling related disorders.

Synthesis, Anti-inflammatory, and Antioxidant Activity of Mannich Bases of Dehydrozingerone Derivatives

Objective: This study aims to synthesize a series of five new Mannich bases of dehydrozingerone (DHZ) derivatives and to evaluate for their antiinflammatory and antioxidant activity. Methods: The synthesis was performed by refluxing DHZ with formaldehyde and secondary amines, and the structures of the synthesized compounds were confirmed by FT-IR, 1 H-NMR, 13 C-NMR, and HR-MS. The anti-inflammatory and antioxidant activity evaluations were done by inhibition of heat-induced albumin denaturation and a free-radical DPPH method, respectively. Results: All the synthesized compounds ( 2a-e ) showed anti-inflammatory and antioxidant activity. The highest anti-inflammatory activity was shown by compound 2c . The activity was comparable to the that of diclofenac sodium as a standard. While, the highest antioxidant activity was demostrated by compound 2e . The compound showed moderate activity compared to that of quercetin as a standard. Mostly of Mannich base derivatives of DHZ compounds exhibited higher antioxidant activity than that of DHZ. Conclusion: A series of five new Mannich bases of DHZ ( 2a-e ) was synthesized successfully. Compound 2c demonstrated anti-inflammatory activity which was comparable to diclofenac sodium, while compound 2e exhibited moderate antioxidant activity compared to quercetin as standard. Key words: Dehydrozingerone, Mannich Bases, Synthesis, Anti-inflammatory, Antioxidant.

PHYTOCHEMICAL ANALYSIS AND SUPPRESSION OF INFLAMMATORY TARGETS BY ADATHODA VASICA

Objectives: A persistent and chronic inflammation may act as an underlying definition for many of the long-term illness such as diabetic foot ulcer, Alzheimer’s disease and rheumatoid arthritis. Folklore medicinal plants provide immense health benefits with new remedies. Active compounds from plants containing alkaloids and phenolic compounds (flavonoids and tannins) possess antioxidant and anti-inflammatory activity. The leaves of Adhatoda vasica are most known for its effectiveness in treating respiratory diseases such as asthma, chronic bronchitis, and other respiratory conditions. In this report, we investigated the possible action of A. vasica against inflammation.Methods: Solvents with varying polarity (Hexane, dichloromethane, ethyl acetate, and methanol) were used for the sequential extraction of leaves of A. vasica Linn (Family: Acanthaceae). The anti‐inflammatory activity was assessed using simple in vitro methods. Phenols, flavonoids, phytosterols, saponins, alkaloids, and tannins were analyzed qualitatively using standard protocols. Anti‐inflammatory activity was evaluated using albumin denaturation assay, membrane stabilization assay and proteinase inhibitory activity. Aspirin (diclofenac) was used as a standard drug for the study of anti-inflammatory activity.Results: Our results substantiated that methanolic extract of A. vasica gave a maximum yield of 12.5% and apparently all bioactive phytochemical constituents were at a higher concentration in the methanolic extract. Methanolic extract of A. vasica showed a significant inhibition of the heat-induced albumin denaturation, red blood cells membrane stabilization, and proteinase inhibitory effects with 86±1.46, 88±1.24, and 96±0.93% for 1000 μg/ml, respectively.Conclusion: The above results delineate that the bioactive components were extracted using high polar solvent are mostly flavonoids and related polyphenols. These compounds may be the active constituents to render the anti-inflammatory activity to A. vasica.

Investigations on anti-inflammatory and analgesic activities of Alnus nitida Spach (Endl). stem bark in Sprague Dawley rats

Ethnopharmacological relevanceStem bark of Alnus nitida (Spach) Endl. (family Betulaceae) is used by local communities in northern areas of Pakistan as a remedy for swelling, injuries and pain. However no pharmacological study of this plant has been reported to confirm these activities. In this study anti-inflammatory and analgesic effect of A. nitida stem bark have been evaluated. MethodsPowder of the stem bark of A. nitida was extracted with methanol (ANBM) and partitioned in escalating polarity to get the n-hexane (ANBH), chloroform (ANBC), ethyl acetate (ANBE) and the residual soluble aqueous (ANBA) fractions. The methanol extract and derived fractions were evaluated for anti-inflammatory activity by using in vitro heat induced albumin denaturation assay and various in vivo assays; carrageenan-induced hind paw edema method, Freunds’ complete adjuvant induced arthritis, histamine induced paw edema and xylene induced ear edema in Sprague Dawley rat. The extracts/fractions were also evaluated for analgesic effects by using hot plate analgesic test and acetic acid induced writhing test in rat. The ANBM composition was analyzed by HPLC-DAD and GC-MS analysis. ResultsResults of heat induced albumin denaturation activity indicated that among the extract/fractions ANBC at concentration range of 100−500µg/ml remarkably protected the heat induced albumin denaturation. The pretreatment with ANBC significantly reduced the carrageenan induced edema with 90.81±1.6% after 4h, comparing with 86.63±3.42% reduction produced by the reference drug diclofenac potassium. Histopathological alterations of the gastric and hind paw were decreased with the extract/fractions. Furthermore, anti-inflammatory effects of ANBC were evident in Freunds’ complete adjuvant induced arthritis, histamine induced paw edema and xylene induced ear edema. The latency time in hot plate analgesic assay with ANBC (61.59±0.38%) after 90min was comparable to standard drug morphine (69.31±2.67%) and aspirin (67.24±2.08%). Similarly ANBC significantly (p<0.01) inhibited the pain sensation in acetic acid induced writhing test in rat. HPLC-DAD analysis of ANBM indicated the presence of gallic acid, catechin and rutin while the GC-MS analysis of ANBM indicated the presence of 30 compounds predominantly of neophytadiene, 3,7,11,15 tetramethyl-2-hexadecen-1-ol, phytol, vitamin E and linalool. ConclusionThe results of this study suggested that the presence of polyphenols, sterols, terpenoids and other constituents might contributed towards the anti-inflammatory and analgesic activities of the crude methanol extract of A. nitida bark and its derived fractions. This study endorsed the folklore use of A. nitida bark for inflammation related disorders.

Exploring the Bioactive Terpenoid Content of an Algerian Plant of the Genus Pulicaria: The ent-Series of Asteriscunolides.

Chemical analysis of the chloroform extract of the aerial parts of the Algerian plant Pulicaria undulata exhibiting anti-inflammatory activity resulted in the isolation of 10 new humulene sesquiterpenoids, 1-10, belonging to the asteriscunolide family of compounds. The structure and relative configuration have been defined by NMR data, whereas the absolute configuration has been established by ECD analysis. Compounds 1-10 include enantiomers of the known asteriscunolides A-D and tetrahydroasteriscunolide previously reported from the genera Asteriscus and Nauplius. Compounds 1 and 10 showed in vitro anti-inflammatory activity by inhibiting heat-induced albumin denaturation with IC50 values of 23.76 and 220.42 μM.

English

This study investigates the anti-inflammatory properties of Hydrocotyle bonariensis Comm. Ex Lam, a medicinal plant used by indigenous traditional healers to manage chronic inflammatory diseases especially rheumatism and arthritis. The hexane extract of H. bonariensis leaves was evaluated for the presence and concentration of phytochemicals. It was subjected to heat-induced albumin denaturation and human red blood cell (HRBC) membrane stabilization assays. The anti-inflammatory properties of the extract were further assessed by employing the formaldehyde induced arthritis animal model assay. The extract was finally subjected to GC/MS analysis for the tentative identification of the phytochemical constituents. Phytochemical analysis of the extract revealed the presence of saponin, phenol, flavonoid, tannin, terpenoid and sterol. This extract showed the ability to inhibit thermally-induced protein denaturation and stabilize HRBC membrane in concentration dependent manner. In the formaldehyde induced arthritis model, the paw measurements of the rats were taken, their hematological parameters were determined, and their liver function tests (aspartate aminotransferase and alanine aminotransferase activities) were also carried out. Results from the animal model indicated that oral administration of the hexane extract of H. bonariensis leaf at a dose of 250 mg/kg body weight had potent anti-inflammatory action. The GC/MS analysis suggested a number of anti-inflammatory compounds in the extract among which were: hexadecanoic acid methyl ester, falcarinol and phytol. Consequently, the tentative identification of phytochemicals with anti-inflammatory activity in H. bonariensis affirms the anti-inflammatory property of the plant and the phytochemicals could serve as lead compounds for designing anti-inflammatory drugs. Key words: Hydrocotyle bonariensis, anti-inflammatory, phytochemicals, protein denaturation, membrane stabilization, arthritis.

Antioxidant and anti-inflammatory activities of Arbutus unedo aqueous extract

ObjectiveTo evaluate the antioxidant and anti-inflammatory activities of aqueous extract of Arbutus unedo (A. unedo) leaves. MethodsIn this context, the in vitro antioxidant activity was demonstrated by 2,2-diphenyl-1-picrylhydrazyl, hydroxyl radical and H2O2 radical scavenging, ferrous ion chelating, ferric reducing power, total antioxidant capacity and by the protection against peroxidation of β-carotene-linoleic acid in emulsion. The anti-inflammatory activity was evaluated first by studying the membrane of human red blood cells against different hypotonic concentrations of NaCl and against heat, inhibiting the denaturation of albumin. ResultsTotal phenolic and flavonoid content were found respectively [(207.84 ± 15.03) mg gallic acid equivalent/g, and (13.070 ± 0.096) mg quercetin equivalent/g]. The extract displayed significant scavenging activity of some radicals such as 2,2-diphenyl-1-picrylhydrazyl [IC50 at (7.956 ± 0.278) μg/mL], OH [IC50 = (1015.74 ± 46.35 μg/mL)], H2O2 [IC50 = (114.77 ± 16.86) μg/mL] and showed a good antioxidant activity through ferrous ion chelating activity [IC50 = (1014.30 ± 36.21) μg/mL], ferric reducing power [IC50 = (156.55 ± 17.40) μg/mL], total antioxidant capacity [IC50 = (461.67 ± 4.16) μg/mL] and β-carotene-linoleic acid protection against peroxidation [I% = (87.04 ± 1.21)% at 1000 μg/mL]. ConclusionsA. unedo showed in vitro anti-inflammatory activity by inhibiting the heat induced albumin denaturation and red blood cells membrane stabilization. Our results show that aqueous leaf extract of A. unedo has good antioxidant activity and interesting anti-inflammatory properties. A. unedo aqueous extract can be used to prevent oxidative and inflammatory processes.