Membrane Stabilization Assay Research Articles

Assessment of Anti-Inflammatory and Antimicrobial Potential of Ethanolic Extract of Woodfordia fruticosa Flowers: GC-MS Analysis.

Currently, the potential utilization of natural plant-derived extracts for medicinal and therapeutic purposes has increased remarkably. The current study, therefore, aimed to assess the antimicrobial and anti-inflammatory activity of modified solvent evaporation-assisted ethanolic extract of Woodfordia fruticosa flowers. For viable use of the extract, qualitative analysis of phytochemicals and their identification was carried out by gas chromatography–mass spectroscopy. Analysis revealed that phenolic (65.62 ± 0.05 mg/g), flavonoid (62.82 ± 0.07 mg/g), and ascorbic acid (52.46 ± 0.1 mg/g) components were present in high amounts, while β-carotene (62.92 ± 0.02 µg/mg) and lycopene (60.42 ± 0.8 µg/mg) were present in lower amounts. The antimicrobial proficiency of modified solvent-assisted extract was evaluated against four pathogenic bacterial and one fungal strain, namely Staphylococcus aureus (MTCC 3160), Klebsiella pneumoniae (MTCC 3384), Pseudomonas aeruginosa (MTCC 2295), and Salmonella typhimurium (MTCC 1254), and Candida albicans (MTCC 183), respectively. The zone of inhibition was comparable to antibiotics streptomycin and amphotericin were used as a positive control for pathogenic bacterial and fungal strains. The extract showed significantly higher (p < 0.05) anti-inflammatory activity during the albumin denaturation assay (43.56–86.59%) and HRBC membrane stabilization assay (43.62–87.69%). The extract showed significantly (p < 0.05) higher DPPH (2,2-diphenyl-1-picrylhydrazyl) scavenging assay and the obtained results are comparable with BHA (butylated hydroxyanisole) and BHT (butylated hydroxytoluene) with percentage inhibitions of 82.46%, 83.34%, and 84.23%, respectively. Therefore, the obtained results concluded that ethanolic extract of Woodfordia fruticosa flowers could be utilized as a magnificent source of phenols used for the manufacturing of value-added food products.

Myristic acid reduces skin inflammation and nociception.

This study evaluated the inhibitory effect of myristic acid (MA) on models of inflammation and nociception. The in vitro anti-inflammatory activities of MA were assessed on LPS-stimulated macrophages, membrane stabilization assay, and inhibition of protein denaturation, whereas the inhibitory activity of MA on in vivo inflammation was assessed on TPA-induced ear edema using acute and chronic assays in mice. The inhibitory effect of MA on nociception was assessed using three in vivo models. MA exerted in vitro anti-inflammatory activity by the increase (58%) in the production of IL-10 in LPS-stimulated macrophages. In the in vivo assay, MA showed good anti-inflammatory effects on the acute (ED50 = 62mg/kg) and chronic (ED50 = 77mg/kg) TPA-induced ear edema. The antinociceptive activity of MA was related to the participation of the nitrergic system in the formalin-induced paw licking test. PRACTICAL APPLICATIONS: Previous studies with different plant extracts containing MA, as one of their major components, have demonstrated anti-inflammatory and antinociceptive actions. However, the anti-inflammatory and antinociceptive actions of myristic acid have not been previously reported. The results suggest that MA induced anti-inflammatory effects in LPS-stimulated macrophages through the participation of IL-10. The antinociceptive effects of MA are attributed to the participation of the nitrergic system.

A search for antiinflammatory therapies: Synthesis, in silico investigation of the mode of action, and in vitro analyses of new quinazolin‐2,4‐dione derivatives targeting phosphodiesterase‐4 enzyme

AbstractInflammation is the response of the body's immune system to harmful stimuli. The expression of phosphodiesterase 4 enzyme (PDE4) was demonstrated in many inflammatory cells. Thus, it was considered an attractive therapeutic target for combating inflammatory disorders. In the present study, a novel class of quinazolin‐2,4‐dione analogues 1‐17a,b was synthesized. Structures of the newly synthesized compounds were characterized by means of spectral and elemental analysis. Additionally, molecular docking studies for the new synthetic compounds were performed using PyRx—virtual screening tool to demonstrate the possible binding pattern mode of interactions within the active site region of the PDE4 enzyme. The antiinflammatory activity of the synthesized compounds was also in vitro examined using inhibition of protein denaturation, anti‐proteinase effect, and membrane stabilization assay. The in silico docking studies revealed that the newly synthesized compounds were well accommodated by the active site region of the target protein through a network of noncovalent interactions such as hydrogen bonds and pi‐stacking, which contributed to the enhancement of affinity and stability between the compounds and the target protein. They exhibited better docking scores when compared with reference drugs diclofenac and coumarin. In addition, in vitro testing revealed that the compounds had moderate to good antiinflammatory effects. The biological study agreed with in silico approach, which revealed that the compounds had promising antiinflammatory activity. Hence, our detailed results can facilitate the rational drug design targeting PDE4 enzyme.

Gloeothece sp.-Exploiting a New Source of Antioxidant, Anti-Inflammatory, and Antitumor Agents.

Bioactive lipidic compounds of microalgae, such as polyunsaturated fatty acids (PUFA) and carotenoids, can avoid or treat oxidation-associated conditions and diseases like inflammation or cancer. This study aimed to assess the bioactive potential of lipidic extracts obtained from Gloeothece sp.–using Generally Recognized as Safe (GRAS) solvents like ethanol, acetone, hexane:isopropanol (3:2) (HI) and ethyl lactate. The bioactive potential of extracts was assessed in terms of antioxidant (ABTS•+, DPPH•, •NO and O2•assays), anti-inflammatory (HRBC membrane stabilization and Cox-2 screening assay), and antitumor capacity (death by TUNEL, and anti-proliferative by BrdU incorporation assay in AGS cancer cells); while its composition was characterized in terms of carotenoids and fatty acids, by HPLC-DAD and GC-FID methods, respectively. Results revealed a chemopreventive potential of the HI extract owing to its ability to: (I) scavenge -NO• radical (IC50, 1258 ± 0.353 µg·mL−1); (II) inhibit 50% of COX-2 expression at 130.2 ± 7.4 µg·mL−1; (III) protect 61.6 ± 9.2% of lysosomes from heat damage, and (IV) induce AGS cell death by 4.2-fold and avoid its proliferation up to 40% in a concentration of 23.2 ± 1.9 µg·mL−1. Hence, Gloeothece sp. extracts, namely HI, were revealed to have the potential to be used for nutraceutical purposes.

STUDY OF ANTI-INFLAMMATORY POTENTIAL OF WHOLE CELL EXRACTS OF CYANOBACTERIA BY MEMBRANE STABILIZATION &amp; TRYPSIN INHIBITION ASSAY

Inflammation is a complex mechanism in response to any infection, injury, or irritation. The prolonged inflammation leads to tissue damage and loss of function result in various disease conditions like osteoarthritis, and autoimmune diseases like lupus, rheumatoid arthritis, asthma and Crohn’s disease. Anti-inflammatory drugs are used to control inflammatory response and prevent tissue damage .Cyanobacteria are rich sources of phytochemicals like phenolics, flavonoids are known to have anti-inflammatory activity. Because of ease of cultivation and faster growth rate than plants, they are preferred candidates over plants. This study focuses on screening of cyanobacterial isolates for their anti-inflammatory activity. The human erythrocytes (HRBC) membrane stabilization assay indicated the potential of whole cell extracts of cyanobacteria to stabilize lysosomal membrane and thereby prevent tissue damage by lysosomal chemokines and enzymes. The trypsin inhibition assay is an indicator for potential to inhibit proteinases and decelerate tissue damage. The whole cell extracts of 10 cyanobacterial isolates of different genus namely Weistellopsis, Pseudophormidium, Oscillatoria, Nostoc, Phormidium, Chlorella, Hapalosiphon under study showed 85%-94% membrane stabilization in hypotonicity induced hemolysis and 92% to 97% membrane stabilization in heat induced hemolysis. The test extracts also showed 48% to 52% inhibition of trypsin. Thus, the isolates under study have application as anti-inflammatory agent.

Screening for anti-inflammatory activity of extract and fractions of Morinda lucida stem bark and detection of bioactive secondary metabolite

Morinda lucida Benth (Rubiaceae) is a versatile plant used in traditional medicine for the treatment of a variety of ailments and the claims of its efficacy are particularly remarkable in the treatment of infections and immuno-inflammatory disorders. This study evaluates the anti-inflammatory properties of methanolic stem bark extract and fractions of M. lucida and also identifies the phytochemicals responsible for its anti-inflammatory activity. The crude extract was subjected to liquid- liquid partitioning successively with n- hexane, ethylacetate, butanol and water. High performance liquid chromatography (HPLC) of the four fractions and Vacuum Liquid Chromatography fraction (VLC) of the promising fraction was evaluated. The effect of the fractions on egg albumen induced rat paw oedema were also evaluated. Anti-inflammatory activity of the fractions was further screened using xylene induce ear oedema models and human red blood cell membrane stabilization test. Ulcerogenic test on the normal stomach mucosa was also evaluated. The result of the egg albumen induced rat paw oedema showed that the butanol fractions maximally inhibited egg albumen induced effect at 400 mg/kg (70%) and 200mg/kg (67.5%) after 180 minutes compared to the positive control, ibuprofen (20mg/kg) with 100% inhibition after 180 minutes. The result of the xylene induced ear oedema showed that the inhibition produced by 100 µg/ear of the Butanol fraction (BF) was 56.67 % and was greater than inhibition produced by 200 µg/ear of ibuprofen (38.89 %). HPLC analysis of the fractions revealed the following phytocompounds; Cytreo- a-pyrone, Cytosporin- J and Waol A. Ulcerogenic test was negative at the doses of 200 mg/kg and 400 mg/kg of the fractions when compare with the indomethacin (positive control) at dose of 50 mg/kg. Human red blood cell membrane stabilization assay showed that BF-VLC 2 (Dichloromethane: methanol (8:2) VLC of Butanol fraction) exhibited concentration dependent inhibition of heat-induced haemolysis while other extract showed a non- concentration dependent inhibition of haemolysis when compared to the standard, ibuprofen. These findings suggest that the stem bark of M. lucida possess promising anti-inflammatory phytocompounds which justify its use in ethno-medicine.

Venlafaxine demonstrated anti-arthritic activity possibly through down regulation of TNF-α, IL-6, IL-1β, and COX-2.

Venlafaxine is a serotonin-norepinephrine reuptake inhibitor used to treat depression. Previous studies demonstrated its anti-nociceptive and anti-inflammatory activities through the suppression of pro-inflammatory cytokines. Present research aimed to explore its anti-arthritic potential. Different in-vitro assays including egg albumin, bovine serum albumin denaturation and human red blood cell (RBC) membrane stabilization assays along with in-vivo models of formaldehyde and complete Freund's adjuvant-induced arthritis were used to study its anti-arthritic effect. Venlafaxine inhibited egg albumin and bovine serum albumin denaturation and preserve the integrity of red blood cells membrane in concentration-dependent manner. In formaldehyde-induced arthritis venlafaxine significantly (p < 0.001) reduced the paw edema on treatment for 10 days. Chronic administration of venlafaxine for 28days in Freund's adjuvant-induced arthritis model decreased the paw volume (p < 0.001), arthritic index (p < 0.01), flexion pain score (p < 0.05), mobility score (p < 0.05), and improved the stance score (p < 0.05). Venlafaxine also significantly declined the rheumatoid factor (p < 0.01) and C-reactive protein (p < 0.05) levels and increased the RBC count (p < 0.01) and Hb value (p < 0.001). Upon PCR analysis venlafaxine remarkably turndown the mRNA expression of TNF-α, IL-6, IL-1β, and COX-2. Taken together it is inferred from current findings that venlafaxine possesses the significant anti-arthritic activity and could be a potential therapeutic option for the treatment of rheumatoid arthritis.

Characterization of Citrusnobilis Peel Methanolic Extract for Antioxidant, Antimicrobial, and Anti-Inflammatory Activity.

Currently, the potential utilization of fruits and vegetable waste as a source of micronutrients and antioxidants has increased. The present study, therefore, aimed to determine the antimicrobial and anti-inflammatory activities of Citrus nobilis peel extract. A modified solvent evaporation technique was employed for peel extract preparation. For effective utilization of the natural product, quantitative analysis of phenolic compounds was carried out using liquid chromatography and mass spectroscopy technique. Phenolic and flavonoids were present in high amounts, while β-carotene and lycopene were present in vestigial amounts. The antimicrobial efficiency of peel extract was evaluated against four bacterial strains including Staphylococcus aureus (MTCC 3160), Klebsiella pneumoniae (MTCC 3384), Pseudomonas aeruginosa (MTCC 2295), and Salmonella typhimurium (MTCC 1254), and one fungal strain Candida albicans (MTCC 183), and zone of inhibition was comparable to the positive control streptomycin and amphotericin B, respectively. The extract of Citrus nobilis peels showed effective anti-inflammatory activity during human red blood cell membrane stabilization (HRBC) and albumin denaturation assay. The extracts also exhibited 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity ranging from 53.46 to 81.13%. Therefore, the obtained results suggest that Citrus nobilis peel could be used as an excellent source of polyphenols and transformed into value-added products.

Effects of Various Phytochemical Fractions of Mundulea sericea on Free Radical Scavenging and Inhibition of Inflammatory Agents

Antioxident and antiinflammatory activities of Mundulea sericea (Willd.) A. Chev. is a medicinally important species belong to the family Fabaceae. This species contains various phytocompounds of entire parts which act as antimicrobial activity, anti-inflammatory activity, anticancer activity, antidiabetic activity, etc. Hence, the present study was aimed to analysis of phytochemical constitutes through Fourier transform infrared (FTIR) and gas chromatography mass spectrometry (GCMS) analysis by six various solvent extracts of hexane (MSBH), petroleum ether (MSBPE), chloroform (MSBC), ethyl acetate (MSBEA), ethanol (MSBE), and methanol (MSBM). The result of phytochemical analysis reported that the MSBE and MSBM extracts are contain more phytochemical constitutes than other extracts. In FTIR and GCMS study, MSBE and MSBM extracts are reported various phytochemical constitutes and out that many bioactive compounds are present of these extracts. Further, MSBE extract was subjected antioxidant and anti-inflammatory activities. In antioxidant activity, four different type assays such as 2,2-diphenyl-1-picryl-hydrazyl-hydrate radical scavenging, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonate), hydrogen peroxide, and nitric oxide (NO) were used. Among this antioxidant study, NO was reported 74.63 inhibition percentage of free radical than other assays. In anti-inflammatory activity, albumin denaturation and human red blood cell membrane stabilization assay were performed by MSBE extract which result reported that the MSBE extract has good anti-inflammatory activity with more than 60% inhibition capacity on both assays. From this study, M. sericea bark extracts contain highly therapeutic phytocompounds which compounds possess good antioxidant and inflammatory activities.

In vitro antidiabetic, anti-inflammatory and antioxidant potential of the ethanol extract of Uromastyx hardwickii skin

Purpose: To evaluate the in vitro antidiabetic, anti-inflammatory and antioxidant potential of the ethanol extract of Uromastyx hardwickii Skin (UHSEE).&#x0D; Methods: The in vitro effects of UHSEE at various concentrations (10 - 250 µg/mL) on the activities of ߙ-amylase, ߙ-glucosidase and glucose uptake by yeast cells were used to evaluate its antidiabetic potential. Nitric oxide (NO), 2,2-diphenyl-1-picrylhydrazyl (DPPH) and hydrogen peroxide inhibitory assay were employed to determine its antioxidant effects, while the anti-inflammatory effects were evaluated using human red blood cell (HRBC) membrane stabilization assay.&#x0D; Results: UHSEE inhibited ߙ-amylase and ߙ-glucosidase enzymes but increased glucose uptake by yeast cells in a concentration-dependent manner (p &lt; 0.05). It also inhibited NO, DPPH, hydrogen peroxide and HRBC hemolysis in a concentration-dependent manner (p &lt; 0.05).&#x0D; Conclusion: Uromastyx hardwickii skin exhibits promising good antidiabetic, antioxidant and antiinflammatory properties in vitro. However, its true potentials in this regard needs to be evaluted in vivo.

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.

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.

In Vitro Anti-Inflammatory Activity of the Root of Thottea siliquosa (Lam.) Rottb., a Medicinal Undershrub in Western Ghats, India

Inflammation can be considered as an essential immune response that helps the body to survive during the conditions of infection and injury. Drugs for curing inflammation had their origin in the accidental discovery of certain plants and their extracts being used for relief of pain, fever and anti-inflammation. Since the plants are considered as the first source of remedies for ailments in human history, a large number of plants were used as drugs against all kinds of ailments through all ages. Thottea siliquosa, commonly known as ‘Alpam’ belongs to the family Aristolochiaceae is a medicinal undershrub, the crude root extracts of which are widely used in traditional tribal healthcare systems in the slopes of the Western Ghats, India, for treating inflammatory responses. The present study is an attempt to scientifically validate this ethnic practice. Root extracts of T. siliquosa were prepared using solvents like chloroform, ethyl acetate, 70% ethanol, methanol, and distilled water by soxhlet mode of extraction. Anti-inflammatory responses in different solvent extracts were analyzed by albumin denaturation assay, anti-protease assay, Human red blood cell membrane stabilization assay (HRBC method), Cyclooxygenase (COX), and Lipoxygenase (LOX) assays. RAW 264.7 cell lines were used for the COX and LOX study. The results illustrate the antiinflammatory potential of the root extracts. The studies on cell lines clearly indicated that the higher anti-inflammatory potential of chloroform extract over other solvent extracts. In tune with the ethno-botanic information, the aqueous extract offered a dose-dependent anti-inflammation property. Since most of the bio activity of plant extracts are contributed by the phytochemicals present in it, further efforts shall be made for the isolation and purification of bioactive phyto components responsible for the anti-inflammation property of the T. siliquosa root.

Sauropus androgynus (L.) leaf phytochemical activated biocompatible zinc oxide nanoparticles: An antineoplastic agent against human triple negative breast cancer and a potent nanocatalyst for dye degradation

The present study delineates the biosynthesis of ZnOSA NPs by availing S. androgynous (L.) aqueous leaf extract. The formation of ZnOSA NPs was substantiated by FT-IR and UV–visible spectroscopy which was further corroborated by the elemental composition study through EDS spectroscopy. The FE-SEM and HRTEM micrographs have revealed the formation of NPs with different shapes and sizes ranging from 12 to 23 nm. The BET isotherms obtained have manifested the mesoporous nature of ZnOSA NPs. The in-vitro cytotoxicity studies of ZnOSA NPs carried out on MDAMB468 human triple-negative breast cancer (TNBC) and NIH3T3 mouse fibroblast cells have evinced their potential cytotoxic effect (75.98 ± 1.07) on cancer cells sparing the normal cells. Further, the results of the AO-EB dual staining assay indicated early apoptosis in TNBC cells by displaying greenish yellow-fluorescence in the nuclei at a IC50 concentration of 53.79 µg/mL of NPs. Additionally; the human erythrocyte membrane stabilization assay has corroborated the haemocompatibility of ZnOSA NPs by exhibiting a % HRBCMS up to 89.15 ± 0.02%. Furthermore, the photocatalytic experiments performed on Methylene Blue dye have revealed the excellent degradation efficiencies of ZnOSA NPs up to 88.01% with high stability and reusability tested during 4 catalytic cycles. Thus, the obtained results have shown greater potential in the anticancer and photocatalytic dye degradation activities of bio-fabricated ZnOSA NPs.

Anti-Inflammatory Activity of the Stem Bark Methanol Extract of Picralima nitida

Aims: To investigate the anti-inflammatory activity of Picralima nitida stem bark methanol extract (PNSBE).&#x0D; Study Design: The study was carried out using in vivo (carrageenan-induced paw oedema and egg albumin-induced paw oedema) and in vitro (Human red blood cell (HRBC) membrane stabilization assay) models in rat.&#x0D; Place and Duration of Study: Department of Veterinary Physiology and Pharmacology, Michael Okpara University of Agriculture, Umudike, Abia State, Nigeria from March to July, 2020.&#x0D; Methodology: The extract was used at the doses of 100, 200 and 400 mg/kg while diclofenac (20 mg/kg) was used as the standard reference drug for the in vivo study (carrageenan-induced paw oedema and egg albumin). For the in vitro study (red blood cell haemolysis), the extract was used at the concentrations of 25, 50, 100, 200 and 400 µg/ml while diclofenac 250 µg/ml was used.&#x0D; Results: In the carrageenan-induced paw oedema model, the extract at the doses used and the reference drug significantly (p &lt; 0.05) reduced the paw edema in the rats at 1 hour in a dose dependent manner. Also, in the egg albumin model, the paw oedema of treated rats was significantly (P &lt; 0.05) reduced by PNSBE dose dependently reducing the mean rat paw oedema from 0.99 ± 0.12 to 0.61 ± 0.06 at the first hour. There was also a concentration dependent inhibition of red blood cell haemolysis by the extract in the HRBC membrane stabilization assay.&#x0D; Conclusion: Picralma nitida demonstrated a significant anti-inflammatory activity in this study.

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.

Development and Evaluation of a Novel Herbal Emulgel for Potential Anti-Inflammatory and Antioxidant Activities &lt;em&gt;in Vitro&lt;/em&gt;

Emulgel is a novel topical drug delivery system which can overcome the major limitation of gel; limited delivery of hydrophobic drugs. Leea indica (Burm.f.) Merr. (Burulla) is a medicinal plant used in Sri Lankan Ayurvedic medicine for many therapeutic purposes. The present study was aimed to develop a novel herbal emulgel using Leea indica leaf extract and evaluate in vitro anti-inflammatory and antioxidant activities. Different formulations (Fg1-Fg5) were prepared by incorporating freeze dried powder of 70% aqueous acetone leaf extract. All formulations (Fg1-Fg5) were tested for their physicochemical stability parameters such as pH, appearance, odor, homogeneity, spread ability, phase separation and washability for 60 days and they were subjected for in vitro anti-inflammatory activity by Human Red Blood Cell membrane stabilization assay and in vitro antioxidant activity by 2, 2-diphenyl-1-picrylhydrazyl (DPPH) assay respectively. Two commercially available gels were used as the positive control. Among the formulated herbal emulgels, Fg5 showed the significantly high values of in vitro anti-inflammatory and antioxidant activities compared to the tested positive controls. At the concentration of 12.5 mg/mL, Fg5 showed percentage inhibition of 93.73 ± 0.90% of heat induced hemolysis of erythrocytes and in vitro antioxidant activity of Fg5 was 3.39 ± 0.01 mmol Trolox per 100 g of emulgel. All formulated emulgels were found to be semi-solid, homogenous, and washable with water, having no phase separation and pH of the formulations were reported in the range of 6 to 7.There was no remarkable variation ofstability parameters tested during 60 days. It isconcluded that the formulated herbal emulgelshave promising anti-inflammatory andantioxidant activities and formulation Fg5 canbe recommended for further investigationsusing in vivo models.

Chemical characterization and anti-arthritic appraisal of Monotheca buxifolia methanolic extract in Complete Freund's Adjuvant-induced arthritis in Wistar rats.

The current study was designed to evaluate the anti-oxidant and anti-arthritic potential of a traditionally used herb, Monotheca buxifolia. The M. buxifolia methanolic extract (MBME) was prepared from the aerial parts of the plant followed by chemical characterization with GC-MS. The anti-oxidant potential of the MBME was demonstrated by DPPH scavenging activity. The effects of MBME on protein denaturation and membrane stabilization were determined by inhibition of egg albumin denaturation and RBC membrane stabilization assays, respectively. The in vivo anti-arthritic potential of the MBME at 50, 100, and 150mg/kg/day was evaluated in Complete Freund's Adjuvant-induced polyarthritis in Wistar rats treated for 21days. Phytochemicals, such as linolenic acid methyl ester, n-hexadecanoic acid, vitamin E, α-amyrin, and β-amyrin were detected in the GC-MS analysis. The plant extract exhibited a 55.20 ± 0.69% scavenging of free radicals at 100µg/ml concentration. It significantly (p < 0.05) stabilized human RBC membrane (65.06 ± 0.22%) and inhibited protein denaturation (70.53 ± 0.34%) at 100mg/ml concentration. The diclofenac sodium (DS) and MBME at 150,100, and 50mg/kg reduced the paw edema, restored the body weight, and altered blood parameters including CRP. The MBME significantly reduced the MDA and increased the SOD, CAT, and GSH levels in liver tissue homogenate in treated rats. The serum concentration of TNF-α and PGE2 was remarkably (p < 0.01-< 0.0001) restored by the DS and MBME dose dependently. The histopathological study showed that MBME 150mg/kg commendably restored the ankle joint inflammation, bone erosion, and cartilage damage in polyarthritic rats. It was concluded that the anti-oxidant, anti-inflammatory and anti-arthritic effects of MBME might be attributed to phenols, flavonoids, triterpenoids, vitamin E, phytol, and other fatty acids. This study showed the anti-arthritic potential of Monotheca buxifolia and thus validates its traditional claim.

Chemical Composition of Pinus brutia Ten Essential Oil and Its in vitro Anti-Inflammatory Activity

This investigation aims to determine the chemical composition of Pinus brutia leaves essential oil and evaluate its anti-inflammatory property using Human Red Blood Cells (HRBC) membrane stabilization assay and Albumin denaturation assay. The chemical composition of essential oil (EO) obtained by hydro-distillation of leaves of Pinus brutia was investigated by GC-MS. The anti-inflammatory effect of EO was evaluated using Human Red Blood Cells (HRBC) membrane stabilization assay and Albumin denaturation assay. The main constituents of EO were α-Terpineol (66.16%), 3-Carene (4.90%), Carveol (4.55%) and cis-Verbenol (3.22%). The inhibion of hemolysis was observed at concentrations (2.5-12.5) µg/ml. Moreover, albumin denaturation test showed protection effect at concentrations (8-40) µg/ml. We concluded that, Pinusbrutia EO shows strong anti-inflammatory activity at different concentration when compared to standard drug of Diclofenac sodium. In addition, GC-MS analysis of Pinus brutia EO showed the presence of α-Terpineol as major compound in the oil. It reveals that this constituent is responsible to maximum protection of albumin denaturation and membrane stabilization assay. The future work will be determination of anti-inflammatory by in vivo models.

Antioxidant and anti-inflammatory potential, and chemical composition of fractions of ethanol extract of Annona muricata leaf

Serious health challenges have been associated with inflammation which is a major cause of mortality in the world. This study evaluated the antioxidant, anti-inflammatory potential, and chemical compositions of fractions of ethanol extract of Annona muricata leaf. The leaves were dried at room temperature, blended and extracted in sequential with solvents of varying degree of polarities, i.e., n-hexane, ethyl acetate and ethanol. Ethanol extract was fractionated via solvent-solvent partitioning into five fractions, i.e., n-hexane fraction (F1), dichloromethane fraction (F2), dichloromethane/ methanol (1:1) fraction (F3), methanol fraction (F4), and ethanol fraction (F5). These fractions were examined for their in-vitro antioxidant activities on DPPH, ABTS and H2O2 while the anti-inflammatory activities were investigated using lipoxygenase inhibition, proteinase inhibition and membrane stabilization assays. The F4 being the most active fraction was further analyzed with GCMS to determine its chemical compositions. The results showed that F4 had the highest H2O2 scavenging activity at 10–100 µg/mL. The activity of F4 at 50 µg/mL was significantly higher (P<0.05) than that of other treatments including the standard (Vitamin C). Activity of F4 also showed significantly higher (P<0.05) membrane stabilization than other fractions at 50-100 µg/mL. F4 exhibited higher antioxidant and anti-inflammatory activities than the other fractions. The activity of this fraction could be attributed to the synergetic effect of various antioxidant compounds present in the fraction. Some of the bioactive compounds identified in the GC-MS of F4 were coumaran, tyrosol, phytol, tetracosanol, elaidic acid methyl ester and β-sitosterol. Keywords: Bioactivity, concentration, inhibition, radical scavenging