Hypotonicity-induced Hemolysis Research Articles

Phytochemical Profiling and Pharmacological Evaluation of Methanolic Leaf Extract of C. digyna for Cytotoxic, Anti-inflammatory, Antioxidant, Antiarthritic, and Analgesic Activities.

Caesalpinia digyna (Family: Fabaceae) is traditionally used in Ayurvedic medicine for various medicinal purposes, including as a treatment for wounds, leprosy, skin diseases, fever, diabetes, etc. Although the root and stem of this plant have a significant medicinal value, there was little research on the leaves of this plant. This study aimed to investigate the qualitative phytochemical profile and evaluate the invitro cytotoxic, anti-inflammatory, antioxidant, and antiarthritic activities, as well as the invivo anti-inflammatory and analgesic activities, of C. digyna leaf extract. The methanolic extract of C. digyna leaves was prepared using an ultrasonic-assisted extraction process. Invitro and invivo anti-inflammatory activities were evaluated using the hypotonicity-induced hemolysis and carrageenan-induced paw edema methods, respectively. Additionally, the extract was assessed for invitro DPPH (1, 1-diphenyl-2-picrylhydrazyl) free radical scavenging, antiarthritic (protein denaturation), and invivo analgesic (acetic acid-induced writhing and tail immersion) activities. Brine shrimp lethality bioassay (BSLB) showed moderate cytotoxic activity (LC50 = 2.25 μg/mL) compared with the standard vincristine sulfate (LC50 = 1.61 μg/mL). Invitro, anti-inflammatory activity exhibited 85.13% (IC50 value = 2.51 μg/mL) inhibition of Human Red Blood Cell (HRBC) membrane lysis at a concentration of 2000 μg/mL whereas invivo anti-inflammatory study exerted its maximum effect (p < 0.05) at 400 mg/kg bw dose. This extract also showed significant antioxidant (IC50 = 0.218 μg/mL), antiarthritic (83.61% inhibition) activity, and moderate analgesic effect (p < 0.05) in both methods. These research findings indicated that C. digyna leaves extract has potent antioxidant, analgesic, and anti-inflammatory effects which can be used as a supplementary medication for inflammatory pain-relieving factors. In future, finding the mechanism involved in these effects could have significant impact on clinical science.

Study on Anti-arthritic and Antimicrobial Activity of Leaves of Mukia maderaspatana on Membrane Stabilization Assay and Egg Albumin Assay

Abstract Background and Aim: Arthritis, a chronic inflammatory disorder, and microbial infections substantially affect global health, driving the need for effective natural remedies. This study explores the anti-arthritic and antimicrobial properties of Mukia maderaspatana leaves, traditionally used in folk medicine. Materials and Methods: Anti-inflammatory activity: Assessed using the membrane stabilization assay to evaluate the inhibition of hypotonicity-induced hemolysis in erythrocytes. Anti-arthritic activity: Measured through the egg albumin assay by observing the prevention of protein denaturation. Antimicrobial activity: Evaluated using standard microbial strains to gauge the broad-spectrum antimicrobial effects of M. maderaspatana leaves. Results: Membrane stabilization: Significant inhibition of hypotonicity-induced hemolysis was observed, indicating notable anti-inflammatory activity. Protein denaturation: Reduced denaturation of egg albumin proteins was noted, suggesting potent anti-arthritic properties. Antimicrobial activity: Broad-spectrum antimicrobial effects were demonstrated against standard microbial strains. Conclusion: M. maderaspatana leaves exhibit significant membrane stabilization, reduced protein denaturation, and broad-spectrum antimicrobial effects. These properties indicate their potential as a natural source for developing treatments targeting arthritis and microbial infections.

Membrane Stabilization, PhospholipaseA2, Albumin Denaturation, Protease Inhibition, as Viable Mechanisms for the Anti-Inflammatory Effects of Methanol Extract of Rauvolfia vomitoria Afzel Leaves

Background: The body uses inflammation as a defence mechanism to eliminate harmful stimuli like germs, damaged cells, or irritants and to initiate the healing process. However, the ongoing discovery of several medicinal plants and the testing of their bioactivity to produce information that will assist doctors and patients in making informed decisions prior to employing them have established.&#x0D; Aims: The study valuate the phosphatelipase A2, membrane stabilization, albumin denaturation, protease inhibition, and platelete aggregation activities as viable mechanisms for the anti-inflammatory effects of the methanol extract of Rauvolfia vomitoria Afzel leaf (MERVL).&#x0D; Methodology: The anti-inflammatory effect of (MERVL) was ascertained using the phosphate lipaseA2, membrane stabilization model, albumin denaturation, protease inhibitor, assay.&#x0D; Place and Duration of the Study: Department of Pharmacology Lab, Enugu State of University of Science and Technology Agbani Nigeria, between March 2021 and August 2021.&#x0D; Results: MERVL inhibited hypotonicity-induced haemolysis by 27.14, 41.10 and 65.70%, at the concentration of 0.4, 0.8 and, 1.0 mg/mL respectively.The highest percentage of inhibition (67.70%) was noticed at the highest concentration of the MERVL. These results were almost analogous to the standard drug (indometacin) used as it exhibited concentration dependent inhibition of albumin denaturation. Protease activity was significantly (P &lt; 0.05) increased at all concentrations which follow the similar tendency as standard drug used. The results showed that MERVL has anti-inflammatory activities.

Anti-Inflammatory and Antinociceptive Activity of Herbal Lipospheres of Pentaclethra macrophylla (Fabaceae) Stem Bark Extract

Purpose: Inflammation of various degrees is common among humans. There are associated side effects with orthodox delivery systems and anti-inflammatory agents; hence, the study investigated the characteristics of herbal lipospheres and the anti-inflammatory potency of the lipospheres formulated from Pentaclethra macrophylla with the view to having a drug with a better delivery system and lesser side effects. Methods: Herbal lipospheres were formulated using solidified reverse micellar solutions (SRMS) of P90H and goat fat and characterized for particle size and morphology, pH time dependent analysis, encapsulation efficiency (EE%), and Fourier Transform infrared spectroscopy. The in vitro antinociceptive and anti-inflammatory studies were carried out using membrane stabilization by hypotonicity-induced hemolysis and the determination of anti-platelet aggregatory activity models. The in vivo antinociceptive and anti-inflammatory studies on egg albumin- and formaldehyde-induced arthritis models were conducted. A total white blood cell count and differential blood count were carried out on the rats. Results: The results showed that there was no change in pH for the PM-unloaded lipospheres and 2.5 g of PM-loaded lipospheres from day 1 to day 7, but there was a mild variation in the rest of the formulations. The EE ranged from 35.2% to 94%, increasing according to the drug concentration. The photomicrographs of the lipospheres showed that the particles were spherical in shape. The particle sizes were within the acceptable range for lipospheres. FTIR showed no interaction. In the arthritis study, PM-loaded lipospheres inhibited edema consistently throughout the duration of observation. Inhibition of the membrane increased steadily with an increase in concentration of PM in the lipospheres and the standard drug. The platelet aggregatory inhibition decreased steadily with an increase in concentration of the PM in the lipospheres as well as the standard. The T50 dose of PM had the highest percentage of WBC, and it decreased as the treatment doses increased from T100 to T200. There were no significant differences among the Neutrophil counts of the different groups. Conclusions: The study, therefore, showed that the methanol extract of Pentaclethra macrophylla formed efficient herbal lipospheres with antinociceptive and anti-inflammatory activities.

Anti-Inflammatory Potential of Seasonal Sonoran Propolis Extracts and Some of Their Main Constituents.

Biological properties of Sonoran propolis (SP) are influenced by harvest time. Caborca propolis showed cellular protective capacity against reactive oxygen species, which might be implicated in anti-inflammatory effects. However, the anti-inflammatory activity of SP has not been investigated so far. This study investigated the anti-inflammatory activity of previously characterized seasonal SP extracts (SPE) and some of their main constituents (SPC). The anti-inflammatory activity of SPE and SPC was evaluated by measuring nitric oxide (NO) production, protein denaturation inhibition, heat-induced hemolysis inhibition, and hypotonicity-induced hemolysis inhibition. SPE from spring, autumn, and winter showed a higher cytotoxic effect on RAW 264.7 cells (IC50: 26.6 to 30.2 µg/mL) compared with summer extract (IC50: 49.4 µg/mL). SPE from spring reduced the NO secretion to basal levels at the lowest concentration tested (5 µg/mL). SPE inhibited the protein denaturation by 79% to 100%, and autumn showed the highest inhibitory activity. SPE stabilized erythrocyte membrane against heat-induced and hypotonicity-induced hemolysis in a concentration-dependent manner. Results indicate that the flavonoids chrysin, galangin, and pinocembrin could contribute to the anti-inflammatory activity of SPE and that the harvest time influences such a property. This study presents evidence of SPE pharmacological potential and some of their constituents.

In vitro and in silico investigation of garlic’s (Allium sativum) bioactivity against 15-lipoxygenase mediated inflammopathies

Introduction: Garlic (Allium sativum) is widely used as a flavor-enhancing dietary ingredient and exhibits a wide spectrum of pharmacological effects. This study aimed to investigate the therapeutic effects of aqueous garlic extract to explore the bioactivity against 15-lipoxygenase (15-LOX) mediated inflammopathies. Methods: In this study, the antioxidant (DPPH free radical scavenging assay and reducing power assay), anti-inflammatory (hypotonicity-induced hemolysis assay and 15-LOX inhibition assay) and anticoagulation (serine protease inhibition assay and prothrombin time assay) effects of the aqueous garlic extract were investigated. Furthermore, in silico molecular docking and dynamic simulation analysis of reported small compounds of garlic against 15-LOX1 and 15-LOX2 were performed to figure out the most efficient phytochemical ligands and validate the anti-inflammatory potential. Results: The DPPH scavenging effect and the reducing power of the extract were found with the IC50 of 213.87 ± 1.49 μgmL-1 and EC50 of 124.78 ± 3.39 μgmL-1, respectively. In the hypotonicity-induced hemolysis and 15-LOX inhibition assay, the IC50 values were observed as 147.59 ± 2.98 μgmL-1 and 250.05 ± 8.48 μgmL-1, respectively. The extract inhibited serine protease activity with an IC50 of 301.33 ± 1.31 μgmL-1 and prevented blood coagulation for 10.05 ± 0.35 minutes in prothrombin time assay. The in silico study identified Rhamnetin as a potential 15-LOX1 and 15-LOX2 inhibitor, and it exhibited a stable interaction with the targets throughout the 100 ns dynamic simulation. Conclusion: The findings of this study provide molecular insights into garlic’s medicinal properties as well as its bioactive compounds, which can be potential therapeutic interventions for 15-LOX mediated inflammations.

Veronica rosea biomolecule profiling, antioxidant potential, dermoprotective effect, anti-inflammatory and hemostatic activities and enzyme inhibitory action

• Eight flavonoids were isolated from the ethyl acetate extract. two detected for the first time in the genus veronica • Antioxidant evaluation was evaluated using DPPH, FRAP, and total antioxidant capacity. • Sun protection factor was calculated to estimate the dermoprotective activity • In vitro hemostatic activity was measured of plasma re-calcification time (PRT) • Veronica rosea is able to inhibit heat- and hypotonicity-induced hemolysis, • Veronica rosea present acetyl-butyrylcholinesterases and urease effects. : Veronica rosea (Plantaginacea) has been previously studied from a phytochemical perspective. The present research intends to perform an in-depth investigation to identify more secondary metabolites in this plant and to explore its pharmacological potential. : The ethyl acetate extract (EAE) of V. rosea was analysed with different chromatographic methods. The obtained pure products were analysed using nuclear magnetic resonance (NMR) and mass spectrometry to identify the structures of the bioactive compounds. The therapeutic potential of EAE was investigated by performing several in vitro assays. : An advanced spectral analysis enabled the structures of 8 flavonoids to be elucidated. Considerable amounts of phenolic, flavonoid, tannin and anthocyanin compounds were detected in the following quantities: 161.32±4.29 mg gallic acid equivalents (GAE)/g, 179.48±9.86 mg quercetin equivalents (QE)/g dry extract, 36.44±3.37 mg tannic acid equivalents (TAE)/g dry extract and 7.47±0.45 mg cyanidin 3-glucoside (Cy-3-glu) equivalents/100 g of dry weight (DW), respectively. Strong evidence of the antioxidant activity of the EAE was established with diphenyl-1-picrylhydrazyl (DPPH; 50% inhibitory concentration (IC 50 ) = 16.09±0.2 μg/mL), ferric reducing antioxidant power (FRAP; IC 50 = 6.35±0.019 μg/mL) and total antioxidant capacity (161.41±10.76 mg EAA/mg extract). This antioxidant activity leads to important solar protection (solar protection factor (SPF) of 41.53±0.019 at a dose of 0.2 mg/ml). The EAE exerted a moderate to robust effect on stabilizing the red blood cell membrane compared to diclofenac, as well as haemostatic action, for which plasma recalcification times were significantly shortened. The extract inhibited acetylcholinesterase and butyrylcholinesterase activities with IC 50 = 121.6±03.29 µg/mL and 108.94±4.24 µg/mL, respectively, and significantly inhibited urease activity with an IC 50 = 132.77±4.73 μg/mL. : The present findings indicate that V. rosea might be viewed as a potential source of bioactive molecules to improve the prognosis of certain diseases.

Anti-Cancer and Anti-Inflammatory Activities of a Short Molecule, PS14 Derived from the Virulent Cellulose Binding Domain of Aphanomyces invadans, on Human Laryngeal Epithelial Cells and an In Vivo Zebrafish Embryo Model.

In this study, the anti-cancer and anti-inflammatory activities of PS14, a short peptide derived from the cellulase binding domain of pathogenic fungus, Aphanomyces invadans, have been evaluated, in vitro and in vivo. Bioinformatics analysis of PS14 revealed the physicochemical properties and the web-based predictions, which indicate that PS14 is non-toxic, and it has the potential to elicit anti-cancer and anti-inflammatory activities. These in silico results were experimentally validated through in vitro (L6 or Hep-2 cells) and in vivo (zebrafish embryo or larvae) models. Experimental results showed that PS14 is non-toxic in L6 cells and the zebrafish embryo, and it elicits an antitumor effect Hep-2 cells and zebrafish embryos. Anticancer activity assays, in terms of MTT, trypan blue and LDH assays, showed a dose-dependent inhibitory effect on cell proliferation. Moreover, in the epithelial cancer cells and zebrafish embryos, the peptide challenge (i) caused significant changes in the cytomorphology and induced apoptosis; (ii) triggered ROS generation; and (iii) showed a significant up-regulation of anti-cancer genes including BAX, Caspase 3, Caspase 9 and down-regulation of Bcl-2, in vitro. The anti-inflammatory activity of PS14 was observed in the cell-free in vitro assays for the inhibition of proteinase and lipoxygenase, and heat-induced hemolysis and hypotonicity-induced hemolysis. Together, this study has identified that PS14 has anti-cancer and anti-inflammatory activities, while being non-toxic, in vitro and in vivo. Future experiments can focus on the clinical or pharmacodynamics aspects of PS14.

Green Synthesis and Characterization of ZnO Nanoparticles Using Pelargonium odoratissimum (L.) Aqueous Leaf Extract and Their Antioxidant, Antibacterial and Anti-inflammatory Activities.

Nanoparticles (NPs) exhibit distinct features compared to traditional physico-chemical synthesis and they have many applications in a wide range of fields of life sciences such as surface coating agents, catalysts, food packaging, corrosion protection, environmental remediation, electronics, biomedical and antimicrobial. Green-synthesized metal NPs, mainly from plant sources, have gained a lot of attention due to their intrinsic characteristics like eco-friendliness, rapidity and cost-effectiveness. In this study, zinc oxide (ZnO) NPs have been synthesized employing an aqueous leaf extract of Pelargonium odoratissimum (L.) as a reducing agent; subsequently, the biosynthesized ZnO NPs were characterized by ultraviolet-visible spectroscopy (UV-Vis), dynamic light scattering (DLS), Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM) and energy-dispersive X-ray spectroscopy (EDX), high-resolution transmission electron microscopy (HRTEM) and selected area electron diffraction (SAED). Moreover, aqueous plant leaf extract was subjected to both qualitative and quantitative analysis. Antioxidant activity of ZnO NPs was assessed by DPPH assay, with varying concentrations of ZnO NPs, which revealed scavenging activity with IC50 = 28.11 μg mL−1. Furthermore, the anti-bacterial efficacy of the green synthesized ZnO NPs against four foodborne pathogenic bacterial strains was examined using the disk diffusion assay, and Staphylococcus aureus (ATCC 8095), Pseudomonas aeruginosa (ATCC10662) and Escherichia coli (ATCC 25922) were found to be the most sensitive against biosynthesized ZnO NPs, whereas the least sensitivity was shown by Bacillus cereus (ATCC 13753). The anti-inflammatory effect was also evaluated for both ZnO NPs and the aqueous leaf extract of P. odoratissimum through the human red blood cells (HRBC) membrane stabilization method (MSM) in vitro models which includes hypotonicity-induced hemolysis. A maximum membrane stabilization of ZnO NPs was found to be 95.6% at a dose of 1000 μg mL−1 compared with the standard indomethacin. The results demonstrated that leaf extract of P. odoratissimum is suitable for synthesizing ZnO NPs, with antioxidant, antibacterial as well as superior anti-inflammatory activity by improving the membrane stability of lysosome cells, which have physiological properties similar to erythrocyte membrane cells and have no hemolytic activity. Overall, this study provides biosynthesized ZnO NPs that can be used as a safe alternative to synthetic substances as well as a potential candidate for antioxidants, antibacterial and anti-inflammatory uses in the biomedical and pharmaceutical industries.

Antimicrobial and anti-inflammatory response by two formulations of Jatyadithailam in healing diabetic foot ulcers

Jatyadithailam is a polyherbal formulation used by Ayurvedic practitioners and clinically reportedfor the treatment of inflammation related ailment specifically in non-healing chronic foot ulcers. The study is focused to validate and compare the Ayurvedic Formulary of India (AFI) and Yogagrantha (YG) formulations of Jatyadithailam in vitro, for its antimicrobial and anti-inflammatory potential. Antimicrobial activity of thailam was determined by broth microdilution method for its minimal inhibitory concentration (MIC) and microbicidal activity (MBC/MFC). In vitro anti-inflammatory activity of varying concentrations was determined by assaying albumin denaturation inhibition, membrane stabilization (hypotonicity-induced hemolysis), heat induced hemolysis and antiproteinase activities for both the formulations. Highest bactericidal and fungicidal activity was recorded by AFI formulation of Jatyadithailam showing low MIC values compared to YG formulation. Of all the tested bacterial strains, both the formulations showed great bactericidal effect against Staphylococcus aureus. Both the formulations exhibited significant dose dependent anti-denaturation and anti-proteinase effectively. The thailam also showed marked stabilization of human red blood cell (HRBC) membrane against thermal hemolysis. Our study confirmed AFI formulation of Jatyadithailam possessed better antimicrobial and significant (p<0.05) anti-inflammatory effect. AFI Jatyadithailam was more effective than YG formulation in terms of dose-dependence activity against infection causing microbes and toxic inflammatory mediators. The outcome of the study emphasizes the positive therapeutic potential of Jatyadithailam to combat infectious and inflammatory conditions.

Antioxidant, antidiabetic, anti-inflammatory and anticancer potential of some seaweed extracts

This study aims to evaluate the secondary metabolites, cytotoxicity, antioxidant, antidiabetic, anti-inflammatory, and anticancer activities of four selected seaweeds Padina pavonica, Taonia atomaria, Jania rubens, and Corallina elongata. The maximum value of phenolic content (176.7 ± 6.9 mg gallic acid equivalents/g crude extract) and tannin content (26.5 ± 4.3 mg tannic acid equivalents/g crude extract), was shown in the methanol extract of J. rubens and P. pavonica, respectively. While the ethanol extract of T. atomaria recorded the highest values of total flavonoid content (374.1 ±27.4 mg quercetin equivalents/g crude extract) and total saponins content (30.2 ± 0.7 mg cholesterol equivalents/g crude extract, respectively) compared to other seaweeds. Methanol extract of J. rubens and C. elongata exhibited the greatest content of alkaloids (25.8 ± 4.4 and 22.7 ± 2.6 mg/g d wt.). The methanol extract of P. pavonica established the highest DPPH radical scavenging activity (55.7% ± 0.1) at 50 μg/mL. Among the selected seaweed extracts, the ethanol extract of T. atomaria demonstrated the maximum α-amylase inhibition capacity (66.3% ± 0.0). Methanol extract of J. rubens and P. pavonica species effectively prevented the hypotonicity-induced haemolysis in a concentration-dependent manner compared with the diclofenac potassium as a standard anti-inflammatory drug. In vitro, the reported results also supported the safety and non-toxicity of the four seaweed extracts on WI-38 cell line at lower concentrations. J. rubens and P. pavonica methanol extracts recorded the highest cell growth inhibition of the HeLa Cancer Cell Lines compared to other seaweed extracts and the standard drug Cisplatin.

Effect of ethanolic extract of Stachys pilifera Benth on subacute experimental models of inflammation and some underlying mechanisms.

Background and purpose:This study was designed to evaluate the anti-inflammatory activities of S. pilifera (HESP) in two sub-acute models of inflammation and clarified some possible mechanisms.Experimental approach:Colorimetric methods were used to determine total phenol and flavonoid contents. Carrageenan or formalin-induced rat paw edema (seven days) and multiple application TPA-induced ear edema in mice (9 days) were used. The concentration of IL-1 and TNF-α were measured in the inflamed paw, as well as MDA levels in the serum and liver. Histopathological studies and in vitro anti-inflammatory effects of the extract were also studied using heat-or hypotonicity-induced hemolysis in RBC humans.Findings/Results:Total phenol and flavonoid contents of HESP were 101.35 ± 2.96 mg GAE/g extract and 660.79 ± 10.06 mg RE g extract, respectively. Oral (100 and 200 mg/kg) and topical application (5 mg/ear) of HESP significantly inhibited formalin-induced paw edema and multiple TPA-induced ear edema. The extract also significantly decreased the serum and liver levels of MDA in the carrageenan and formalin tests. The elevated levels of TNF-α and IL-1β in the carrageenan-injected paw were not affected by HESP. The extract (50-800 μg/mL) inhibited heat-or hypotonicity-induced hemolysis. Histopathological examination of the inflamed tissues revealed that HESP inhibited congestion and leukocyte infiltration.Conclusion and implications:The findings confirmed the potent anti-inflammatory effects of S. pilifera in two sub-acute inflammation models and suggested that these properties were not related to IL-1 and TNF-α, but could be attributed to inhibition of lipid peroxidation, membrane stabilization, and inhibition of leukocyte penetration.

INVESTIGATION ON PHARMACOGNOSY AS WELL AS THE ANTIOXIDANT, ANTI-INFLAMMATORY POTENTIAL OF THE KATHA POWDER

Objective: The objective of the study was to investigate the pharmacognosy as well as the antioxidant, anti-inflammatory potential of the Katha powder.&#x0D; Methods: The Coarsely dried chips of Acacia catechu heartwood were treated with 10 % hydro-alcoholic solution to obtain Katha as the final product. The powdered Katha was standardized through pharmacognostic parameters. This Katha power is showing the good solubility in the hot water having astringent in the taste. The powder microscopy of the Katha powder is to be demonstrated fragments of acicular crystals, fibers, and bordered pitted vessels. Katha powder antioxidant potential is to be accessed by using the 2, 2-diphenyl-1-picryl hydrazyl assay and NO Scavenging assay using ascorbic acid as a standard drug. Further, the Katha powder is to be subjected for the assessment of its anti-inflammatory potential by the use of heat-induced hemolysis as well as hypotonicity-induced hemolysis approach by the use of the aspirin or diclofenac sodium as a standard drug.&#x0D; Results: Microscopical investigations were showed that Katha showing the presence of fragments of acicular crystals, fibers, and bordered pitted vessels. In vitro study shows that the Katha powder has excellent antioxidant as well as anti-inflammatory potential in a dose-dependent manner in comparison of the result of heartwood of A. catechu.&#x0D; Conclusion: So from this investigation, it is to be suggested that the Katha powder is rich in the phenolic compound and the experimentation study shows that the drug is to possess a good antioxidant as well as anti-inflammatory property.

Eleusine indica Linn, Baertin (Poaceae) Ethanol Leaf Extract and Its Ethyl Acetate Fraction Display Potential Anti-inflammatory Activities

Objective: Inflammation is the underlying cause of most of the chronic diseases that occur with aging. Although many drugs are available for the management of inflammatory disorders and their symptoms, most of these drugs possess serious adverse effects that limit their usefulness. This has encouraged the unending search for potent anti-inflammatory drugs from plant sources as alternatives to conventional drug treatment of inflammation. This study investigated the anti-inflammatory activities of the ethanol leaf extract of E. indica and the ethylacetate fraction in rodents.&#x0D; Materials and Methods: The leaves were extracted with ethanol by cold maceration and the extract was fractionated with n-hexane, ethylacetate, butanol and water. The oral acute toxicity (LD50) of the extract and the phytochemical constituents of the extract and the fractions were determined. The anti-inflammatory activities of the ethanol extract (EE) and ethylacetate fraction (ETF) and their possible mechanisms of actions were investigated.&#x0D; Results: The oral LD50 of the extract was above 5000 mg/kg. Both the EE and ETF displayed dose-dependent inhibition of the rat paw edema, with ETF producing between 48-54% edema inhibition. Xylene-induced topical edema was significantly (p &lt; 0.05) reduced by both the EE and ETF, with ETF causing between 48 and 65% inhibition. The EE and ETF preserved the integrity of gastric mucosa. Their average ulcer index (1.37±0.02) was significantly lower than that of indomethacin (5.20±0.23). Pre-treatment with the EE and ETF significantly (p &lt; 0.05) reduced leucocyte migration, especially the neutrophils. Both heat- and hypotonicity-induced hemolysis of RBC membrane were remarkably inhibited.&#x0D; Conclusion: The mechanisms of the anti-inflammatory activity may involve among others inhibition of leukocyte migration and membrane stabilization.

IN VITRO ANTI-INFLAMMATORY AND ANTI-ARTHRITIC ACTIVITY OF ETHANOLIC EXTRACT OF LEAVES OF PYRENACANTHA VOLUBILIS (EEPV)

Objectives: The study was undertaken to evaluate the in vitro anti-inflammatory and anti-arthritic activity of the ethanolic extract of leaves of Pyrenacantha volubilis (EEPV) using human red blood cells (HRBCs) membrane stabilization and protein denaturation methods.&#x0D; Methods: In the present study, the in vitro anti-inflammatory and anti-arthritic activity of EEPV was carried out using HRBC membrane stabilization by hypotonicity-induced hemolysis and protein denaturation using egg albumin methods at various concentrations (100, 200, 400, 800, and 1000) of EEPV. Diclofenac sodium was used as reference standard.&#x0D; Results: P. volubilis was effective in inhibiting HRBC membrane stabilization and protein denaturation in a dose-dependent manner and was comparable to the standard drug diclofenac sodium.&#x0D; Conclusion: The study suggests that P. volubilis has potential anti-inflammatory and anti-arthritic activity.

ANTI-INFLAMMATORY PROFILING OF SEED AND RIND EXTRACTS OF AMOMUM SUBULATUM

Objective: The study aimed to evaluate the anti-inflammatory activity of Amomum subulatum (greater cardamom) seed and rind extracts in Wistar rats.&#x0D; Methods: The seed and rind of A. subulatum were air-dried in the shade, powdered, and subjected to 80% hydroalcoholic extraction in the Soxhlet apparatus. The anti-inflammatory activity of the seed and rind extracts of A. subulatum was evaluated by in vivo and in vitro methods.&#x0D; Results and Discussion: In vivo studies, namely, carrageenan-induced rat paw edema, cotton pellet granuloma, and formaldehyde-induced arthritis model confirmed the anti-inflammatory potential of seed and rind extracts of A. subulatum. It was found that rind extract exhibited better inhibition of inflammation as compared to seed extract. A. subulatum rind extract at the dose of 500 μg/ml exhibited best results for in vitro studies, namely, inhibition of albumin denaturation (73% inhibition), antiproteinase action (58% inhibition), membrane stabilization, heat-induced hemolysis, hypotonicity-induced hemolysis (54% inhibition), anti-lipoxygenase activity.&#x0D; Conclusions: The results of the study showed that the rind extract of A. subulatum (greater cardamom) possesses significant anti-inflammatory potential in various in vivo and in vitro anti-inflammatory models in the experimental animals.

In-vitro anti-inflammatory and antioxidant activities of an Ayurvedic formulation –Trayodashang guggulu

Trayodashang guggulu is an important Ayurvedic polyherbal formulation containing babula (stem bark of Acacia nilotica), ashwagandha (roots of Withania somnifera), hapusa (fruits of Juniperus communis), guduchi (stem of Tinospora cordifolia), shatavari (roots of Aspargus recemosus), gokshuara (fruits of Tribulus terrestris), vradadaru (roots of Argyeria nervosa), rasana (roots and leaves of Pluchea lanceolata), satavha (fruits of Anethum sowa), sati (rhizome of Hidichium spicatum), yavani (Trachyspermum ammi fruits), sunthi (rhizome of Zingiber officinale), shuddha officinale guggulu (exudates of Commiphera mukul) and goghrat (ghee). Trayodashang guggulu is used in the Ayurvedic system of medicine for the treatment of various inflammatory conditions such as arthritis and associated pain. Hence, the present study investigated the in-vitro anti-inflammatory and antioxidant activities of trayodashang guggulu. Trayodashang guggulu was standardized as per standard procedures and TLC profile was carried as per Ayurvedic Pharmacopoeia of India. In-vitro anti-inflammatory activity of its aqueous extract (AqTG) (in different concentrations) was evaluated by assaying inhibition of albumin denaturation, membrane stabilization (hypotonicity-induced haemolysis), anti-lipoxygenase and antiproteinase activities. The in-vitro antioxidant effect was evaluated by various in-vitro methods viz. DPPH (1, 1-diphenyl-2-picryl-hydrazyl) and hydroxyl radical scavenging and reducing power assay and total phenolic and flavonoids contents. Trayodashang guggulu was confirmed as per pharmacopoeial standards. It showed marked scavenging effect on DPPH and hydroxyl radicals and exhibited strong reducing potential. It also showed inhibition of membrane stabilizing, protein denaturation inhibitory, anti-lipoxygenase and anti-proteinase activities. Trayodashang guggulu exhibited in-vitro anti-inflammatory and antioxidant activities. This preliminary study supports the therapeutic claim of the formulation as an anti-inflammatory drug in the Ayurvedic system of medicine and advocates its use in inflammatory conditions. In view of the in-vitro results, further in-vivo investigations are needed to confirm and strengthen the anti-inflammatory activity of trayodashang guggulu in pain, inflammation and arthritis.

Extraction and characterization of phycocyanin from Spirulina platensis and evaluation of its anticancer, antidiabetic and antiinflammatory effect

The phycocyanin was purified by Sephadex- G-100 and RP-HPLC and protein content was found to be 52.82% and the high purity fraction was collected and RP-HPLC analysis of fractionated phycocyanin, the α-subunit and β-subunit were detected in 4.9 and 11.1(mAU). The frequency of peak 1456.26 cm−1 has showed the CH2 bending vibration and the protein amide II band was detected at 1539.20 cm−1 (CO stretching) and 2358.94 cm−1. In 1H NMR analysis, 14 chemical shifts (δ) were observed and signals confirmed namely alkyl halide, alkene, aldehyde proton and carboxylic acid. The in vivo anticancer effect was assessed by MTT assay against HepG-2 cell lines and in vivo antidiabetic effect was carried out through α-amylase and β-glucosidase enzyme inhibition methods. The promising anticancer effect 68% was noticed at the concentration of 500 μg/ml and lower anticancer effect was noticed at the concentration of 100 μg/ml against Hep-G2 cell lines. The α-amylase and β-glucosidase enzyme inhibition of phycocyanin showed dose dependent and maximum inhibition effect at 250 μg/ml. Phycocyanin anti-inflammatory effect such as inhibition of albumin denaturation, antiproteinase, hypotonicity-induced haemolysis and anti-lipoxygenase activities have been recorded maximum level at 500 μg/ml. Phycocyanin have complex structure and high molecular weight with more biomedical applications for drug development.

RP-HPLC Estimation of Flavonol Derivatives and Phenolic Acids in Capsicum annuum L. and their Correlation with In Vitro Anti-Inflammatory Activity.

Flavonol derivative and phenolic acids derived from the plants function as free radical scavengers, reducing agents, and quenchers for the formation of singlet oxygen. Flavonoids and phenolic constituents also play an important role in various human diseases and disorders primarily through modulation of inflammatory responses. To estimate the Flavonol Derivatives (FD) and phenolic acids (PA) in Capsicum annuum (CA) and other important phytochemicals having an anti-inflammatory effect. In the present study, FD and PA were estimated in CA and in vitro anti-inflammatory activity (pilot study) was determined and correlation was established. The results were found to be significant using RP-HPLC. FD and PA were found to be 0.0659±0.0058 and 0.0862±0.0.0134 mg/gram dry weight, respectively. For in vitro anti-inflammatory activity, the inhibition of albumin denaturation and antiproteinase activity was found to be maximum in Quercetin (QE) with 98.230±1.589% and 59.906±1.529%, respectively. Heat-induced hemolysis of erythrocytes was found to be maximum in salicylic acid (SA) (71.830±2.838%). Hypotonicity-induced hemolysis showed significant activity with QE (76.770±3.475%). Lipoxygenase and cyclooxygenase inhibition was found to be maximum in QE with 56.930±4.069% and 61.660±3.135%, respectively. A strong positive correlation of 0.9 was observed between the extract of CA and standard QE and SA against the anti-inflammatory activity. Therefore, the role of FD and PA has been postulated to be an active phytochemical of CA accountable for its anti-inflammatory activity. However further work is desirable to fully elucidate the phytochemicals responsible for their anti-inflammatory activity and to develop better herbal drug formulations.

Evaluation of anti-inflammatory activity of Justicia secunda Vahl leaf extract using in vitro and in vivo inflammation models

BackgroundJusticia secunda Vahl. is a medicinal plant used in ethnomedical practice as therapy to manage inflammation. Therefore, this study was designed to evaluate the anti-inflammatory activity of methanol extract of J. secunda leaves (MEJSL) using in vitro and in vivo inflammation models.MethodsSeventy-percent MEJSL was prepared following standard procedure. In vitro anti-inflammatory assays were performed using heat-induced bovine serum albumin (BSA) denaturation and erythrocyte membrane stabilization assays. Carrageenan and formaldehyde induced inflammation in rat models were used to evaluate the anti-inflammatory activity of MEJSL in vivo. Diclofenac sodium was used as a reference drug. In addition, liver and kidney function assays and hematological analysis were carried out.ResultsData revealed that varying concentrations of MEJSL significantly (P < 0.05) inhibited heat-induced BSA denaturation and stabilized erythrocyte membrane against hypotonicity-induced hemolysis when compared with diclofenac sodium in a concentration-dependent manner. In vivo study showed that 10 mg/kg body weight (b.w.) diclofenac sodium, 100 and 300 mg/kg b.w. MEJSL suppressed carrageenan-induced paw edema at the sixth hour by 71.14%, 83.08%, and 89.05%, respectively. Furthermore, 10 mg/kg b.w. diclofenac sodium, 100 and 300 mg/kg b.w. MEJSL inhibited formaldehyde-induced paw edema by 72.53%, 74.73%, and 76.48%, respectively. Animals treated with varying doses of MEJSL had reduced plasma aspartate aminotransferase and alanine aminotransferase activities; urea and creatinine concentrations; and modulated hematological parameters when compared with the untreated control group.ConclusionsFindings from this study showed that MEJSL exhibited substantial anti-inflammatory actions in the in vitro and in vivo models. It also indicated that MEJSL anti-inflammatory mechanisms of action could be through interference with phase 2 inflammatory stressors, upregulation of cytoprotective genes, stabilization of inflammatory cell membranes and immunomodulatory activity.