Inhibition Of Nitric Oxide Production Research Articles

Anti-inflammatory activity via NO production inhibition of compounds from Vietnamese Lycopodium casuarinoides Spring

In our investigation of the methanol extract derived from the aerial parts of Vietnamese Lycopodium casuarinoides Spring, we successfully isolated two novel secondary metabolites: 3β,21α,24-trihydroxyserrat-14-en-29-yl-E-4′-hydroxy-3′-methoxycinnamate (1) and lycocasuarinol A (2), and eight known compounds, namely α-onocerin (3), salcolin A (4), salcolin B (5), (2 R,3 R)-dihydrokaempferol (6), apigenin (7), huperzinine (8), 4-methoxycinnamic acid (9), and (E)-p-coumaric acid (10). The structural elucidation of these compounds was accomplished through high-resolution electrospray ionization mass spectrometry (HR-ESI-MS) and one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR) spectroscopy techniques. All isolates were tested for their potential to inhibit nitric oxide (NO) production in lipopolysaccharide (LPS)-induced RAW264.7 cells. Notably, the flavonoids (2 R,3 R)-dihydrokaempferol (6) and apigenin (7) exhibited significant inhibitory activity against NO production, with IC50 values of 3.4 µM and 3.0 µM, respectively. These values surpassed the inhibition observed with the positive control, celastrol. Furthermore, a comparison of the guaiacyl glyceryls salcolin A (4) and salcolin B (5), differing in their threo-β- and erythro-guaiacyl glyceryl moieties, revealed potential structure-activity relationships. These findings suggest that the extract from Lycopodium casuarinoides and its phytochemical constituents hold promise as potential anti-inflammatory agents

Lanostane triterpenoids from mycelia-associated Ganoderma sinense and their anti-inflammatory activity

Seven previously undescribed lanostane triterpenoids, ganoderic acid M1 (1), M2 (2), M3 (3), M4 (4), M5 (5), M6 (6), and M7 (7), together with eight known compounds, were isolated from mycelia of the basidiomycete Ganoderma sinense (Ganodermataceae). The structures of all compounds were elucidated by spectroscopic analysis. The possible biosynthetic pathway of these fifteen triterpenoids was proposed. Some of the compounds were evaluated for their anti-inflammatory activity by measuring the production of nitric oxide (NO), TNF-α, and IL-6 in RAW264.7 macrophage cells induced by lipopolysaccharide. Lanosta-7,9(11),24-trien-3β,15α,22β-triacetoxy-26-oic acid (14) exhibited the strongest inhibition of NO production with an IC50 of 0.6 ± 0.1 μM and completely inhibited the secretion of TNF-α and IL-6 at 10 μM. The structure-activity relationship of the anti-inflammatory activity is discussed.

A new anti-inflammatory thujane monoterpenoid glycoside ester from Pittosporum heterophyllum Franch

Phytochemical investigation of EtOH extract of Pittosporum heterophyllum led to one new esterified thujane monoterpenoid glycoside, pitheteroside A (1), together with one eudesmane sesquiterpenoid (2) and twelve lignans (3－14). Their structures were elucidated by extensive spectroscopic analysis, including 1D and 2D NMR, ECD calculation and HRESIMS spectra. Pitheteroside A is an unreported and highly esterified monoterpenoid glycoside. All isolates were evaluated for their nitric oxide production inhibition against LPS-induced BV-2 microglial cells. Among them, compounds 1, 6 and 8 showed significant activities with IC50 values less than 10 μM. The results indicated the metabolisms from P. heterophyllum possess potential anti-inflammatory effects.

ANTIOXIDANT AND INHIBITORY EFFECT OF SELECTED GHANAIAN VEGETABLES ON NITRIC OXIDE EXPRESSION IN LIPOPOLYSACCHARIDE-INDUCED MACROPHAGE CELLS

Objective: Nitric oxide (NO) is a signaling molecule that plays a key role in the pathogenesis of inflammation. Inhibitors of NO may be useful candidates for the treatment of inflammatory diseases. The study aimed to determine the antioxidant and inhibitory effect of commonly used Ghanaian vegetables, namely Corchorus olitorius (CO), Solanum melongena (SM), Solanum torvum (ST), Xanthosoma sagittifolia (XS) and Abelmoschus esculentus (AE) on NO expression in a Lipopolysaccharide (LPS)-induced RAW 264.7 macrophage cell line. Methods: The cytotoxic effects of the vegetables on the cell line were determined using a tetrazolium-based colorimetric assay. The inflammatory activity was determined by measuring the inhibition of NO production in LPS-induced RAW 264.7 macrophage cells. Total antioxidant activity, total phenolic, flavonoid, and reduced glutathione contents were evaluated using 2, 2-diphenyl-1-picrylhydrazyl (DPPH) assay, Folin-Ciocalteu, aluminium chloride, and O-Phthalaldehyde methods, respectively. Results: Our results showed that CO and ST significantly inhibited NO production in a concentration-dependent manner with good cell viability. Solanum torvum also exhibited strong antioxidant activity (IC50= 0.466±0.23 mg/ml) with total phenolic content of 230.73±1.84 mg/g GAE, while CO showed high flavonoid content (291.45±2.14 mg/g QUE). Abelmoschus esculentus recorded the highest glutathione content (58.6 µg/g GSH. Saponins, alkaloids, tannins, terpenoids, and cardiac glycosides were present in all the samples except SM and AE, which lacked terpenoids. Conclusion: These findings suggest that CO and ST possess anti-inflammatory and antioxidant activities that could be explored as potential therapeutic remedies for inflammatory disorders.

Establishing Nitric Oxide Levels as a Biomarker for Fibromyalgia and Rheumatoid Arthritis Diagnosis

Fibromyalgia (FM) is a chronic, hyperalgesic disorder leaving musculoskeletal pain throughout the body without tissue damage, ultimately affecting about 1 in 25 to 1 in 50 people in the United States. Although prevalent, its diagnosis is complicated and long due to its ambiguity of official signs of its presence. Patients wait an estimated 2.3 years and see about 3.7 physicians on average before being diagnosed with this disorder. But in recent years, the FM/a blood test, which analyzes unusual patterns of immune responses has allowed for more accurate and timely diagnosis, boasts a 93% sensitivity rate and 89% specificity rate. Although an advancement, the FM/a test was found to have a false positive rate of 29% and 31% for patients with SLE and RA, respectively. To combat the lack of secure physiological ways to determine Fibromyalgia in patients, measuring nitric oxide (NO) levels in the body and exhaled breath appears to be a promising biomarker to distinguish RA from FM upon diagnosis. Nitric oxide works as a vasodilator and signal for localized inflammation. FM is a non-inflammatory disorder, so NO levels are heightened in both the blood and exhaled breath due to a lack of arginase, a competitive inhibitor of NO production. On the other hand, RA is an inflammatory autoimmune disorder that presents similarly in behavior to FM, but it leaves less NO in the blood because of the increased activity of arginase.

Undescribed Phenolic Glycosides from Syzygium attopeuense and Their Inhibition of Nitric Oxide Production.

Four undescribed phenolic glycosides including three stilbene derivatives (1 and 3) and sodium salt of 3 (2), and a chalcone glycoside (4), together with thirteen known compounds (5-17) were isolated from the leaves of Syzygium attopeuense (Gagnep.) Merr. & L.M.Perry. Their chemical structures were elucidated to be (Z)-gaylussacin (1), 6''-O-galloylgaylussacin sodium salt (2), 6''-O-galloylgaylussacin (3), 4'-O-[β-D-glucopyranosyl-(1→6)-glucopyranosyl]oxy-2'-hydroxy-6'-methoxydihydrochalcone (4), gaylussacin (5), pinosilvin 3-O-β-D-glucopyranoside (6), myricetin-3-O-(2''-O-galloyl)-α-L-rhamnopyranoside (7), myricetin-3-O-(3''-O-galloyl)-α-L-rhamnopyranoside (8), myricetin-3-O-α-L-rhamnopyranoside (9), quercitrin (10), myricetin-3-O-β-D-glucopyranoside (11), myricetin-3-O-β-D-galactopyranoside (12), quercetin 3-O-α-L-arabinopyranoside (13), myricetin-3-O-2''-O-galloyl)-α-L-arabinopyranoside (14), (+)-gallocatechin (15), (-)-epigallocatechin (16), and 3,3',4'-trimethoxyellagic acid 4-O-β-D-glucopyranoside (17) by the analysis of HRESIMS, 1D and 2D NMR spectra in comparison with the previously reported data. Compounds 1-3, 5, and 6 significant inhibition of NO production in LPS-activated RAW264.7 cells, with IC50 values ranging from 18.37 ± 1.38 to 35.12 ± 2.53 µM, compared to a positive control (dexamethasone) with an IC50 value of 15.37 ± 1.42 µM.

Biological properties of mucus from land snails (Lissachatina fulica) and freshwater snails (Pomacea canaliculata) and histochemical study of mucous cells in their foot.

Mucus derived from many land snails has been extensively utilised in medicine and cosmetics, but some biological activities of the mucus need to be well documented. Nevertheless, most mucus is obtained from land snails, while mucus from freshwater snails has yet to be attended. This study aims to determine and compare mucus's antioxidant and anti-inflammatory activities from the land snail Lissachatina fulica and the freshwater snail Pomacea canaliculata. ABTS, DPPH, reducing power and total antioxidant activity assays were used to evaluate the antioxidant capacity. Inhibition of nitric oxide production in lipopolysaccharide-activated RAW 264.7 cells was performed to determine the anti-inflammatory activity. Additionally, the histochemical analysis of mucous cells in each snail foot was conducted to compare the distribution of mucous cells and types of mucins using periodic acid-Schiff and Alcian blue staining. Mucus from L. fulica and P. canaliculata exhibited antioxidant and anti-inflammatory activities in different parameters. L. fulica mucus has higher total antioxidant (44.71 ±2.11 mg AAE/g) and nitric oxide inhibitory activities (IC50 = 9.67 ±0.31 µg/ml), whereas P. canaliculata mucus has better-reducing power activity (43.63 ±2.47 mg AAE/g) and protein denaturation inhibition (IC50 = 0.60 ±0.03 mg/ml). Histochemically, both species' dorsal and ventral foot regions contained neutral and acid mucins in different quantities. In the dorsal region, the neutral mucins level in L. fulica (16.64 ±3.46%) was significantly higher than that in P. canaliculata (11.19 ±1.50%), while the acid mucins level showed no significant difference between species. Levels of both mucins in the ventral foot region of L. fulica (15.08 ±3.97% and 10.76 ±3.00%, respectively) were significantly higher than those of P. canaliculata (2.25 ±0.48% and 2.71 ±0.56%, respectively). This study revealed scientific evidence of the biological capacity of mucus from L. fulica and P. canaliculata as well as provided helpful information on the region of the foot which produces effective mucus.

Development of a novel histone deacetylase inhibitor unveils the role of HDAC11 in alleviating depression by inhibition of microglial activation

Histone deacetylases (HDACs) are key epigenetic regulators and classified into four subtypes. Despite the various roles of each HDAC isoform, the lack of selective HDAC inhibitors has limited the elucidation of their roles in biological systems. HDAC11, the sole class-IV HDAC, is highly expressed in the brain, however, the role of HDAC11 in microglia is not fully understood. Based on the modification of MC1568, we developed a novel HDAC inhibitor, 5. Interestingly, 5 suppresses lipopolysaccharide-induced microglial activation by the initiation of autophagy and subsequent inhibition of nitric oxide production. Furthermore, we demonstrated that 5 significantly alleviates depression-like behavior by inhibiting microglial activation in mouse brain. Our discovery reveals that specific pharmacological regulation of HDAC11 induces autophagy and reactive nitrogen species balance in microglia for the first time, which makes HDAC11 a new therapeutic target for depressive disorder.

GC-MS Analysis and Bioactivity Screening of Leaves and Fruits of Zanthoxylum armatum DC.

Zanthoxylum armatum DC. is a plant that has been homologated for medicine and food by the Chinese for three thousand years. In this study, the essential oils of fresh leaves and fruits were extracted by hydrodistillation, the aromas of fresh leaves and fruits were extracted by headspace solid-phase microextraction and their chemical compositions were analyzed by gas chromatography mass spectrometry. The main components of the leaf essential oils were linalool (62.01%), 2-undecanone (9.83%) and 2-tridecanone (5.47%); the fruit essential oils were linalool (72.17%), limonene (8.05%) and sabinene (6.77%); the leaf aromas were limonene (39.15%), β-myrcene (15.8%), sabinene (8.17%) and linalool (5.25%); the fruit aromas were limonene (28.43%), sabinene (13.56%), linalool (11.47%) and β-myrcene (8.64%). By comparison, it was found that the composition of leaf essential oils and fruit essential oils were dominated by oxygenated monoterpenes, while the composition of their aromas were both dominated by monoterpenes; the relative content of non-terpene components in leaf essential oil and leaf aroma is second only to oxygenated monoterpenes, while their content in fruits is low; the chemical composition of leaf aromas and fruit aromas were richer than those of essential oils. In this study, we reported for the first time that the antitumor, tyrosinase inhibition, HMGR inhibition and nitric oxide production inhibition activity of leaf essential oils were stronger than those of fruit essential oils in in vitro tests. The results of the study can provide a reference for the recycling and green low-carbon transformation of the leaves, and also help to deepen the understanding of the value of the volatile chemical constituents of this plant in “forest medicine” or “aromatherapy”, and provide new ideas for the transformation of the value of the plant in the secondary and tertiary industry chain.

Undescribed 2,9-deoxyflavonoids and flavonol-diamide [3+2] adduct from the leaves of Aglaia odorata Lour. Inhibit nitric oxide production

Phytochemical study on the methanol extract of Aglaia odorata leaves resulted in the isolation of four previously undescribed compounds, including three 2,9-deoxyflavonoids and one flavonol-diamide [3 + 2] adduct, and 13 known compounds. The chemical structures of the four undescribed compounds were elucidated on the basis of their IR, HR-ESI-MS, 1D and 2D NMR, and ECD spectra. The results revealed an unprecedented 2,9-deoxyflavonoid framework, which was confirmed by TD-DFT, ECD, and GIAO 13C-NMR calculations using sorted training set methods. The 17 compounds were examined for their ability to inhibit NO production activity in cultured lipopolysaccharide-activated RAW264.7 cells with aglaodoratas A-C, odorine, and epi-odorine inhibiting NO production, with IC50 values in the range of 16.2–24.3 μM. The other investigated compounds had either weak or no activity.

Sesquiterpenoids from the roots of Aucklandia costus and their anti-inflammatory activities

Five undescribed sesquiterpenoid dimers, aucklandiolides A–E (1–5), one new sesquiterpenoid glycoside, β-cyclocostunolide-15-β-D-glucopyranoside (6), and seventeen known analogues (7–23) were isolated from the roots of Aucklandia costus. Their structures were elucidated by comprehensive HRESIMS and NMR spectroscopic data analysis, and their configurations were confirmed by the computational calculations of ECD and NMR chemical shifts. Aucklandiolides A and B are the first examples of dimeric sesquiterpenoids with a unique 6/6/6/5/6/6 ring system originated from a proposed Diels–Alder cycloaddition between two eudesmane sesquiterpenoids. Besides, compounds 9–11, 20, and 22 showed significant inhibition of nitric oxide production in LPS-stimulated RAW 264.7 cells at a concentration of 20 μM.

Datura stramonium Flowers as a Potential Natural Resource of Bioactive Molecules: Identification of Anti-Inflammatory Agents and Molecular Docking Analysis.

The present study investigated the antioxidant, antibacterial, antiviral and anti-inflammatory activities of different aerial parts (flowers, leaves and seeds) of Datura stramonium. The plant material was extracted with 80% methanol for about 24 h. The sensitivity to microorganisms analysis was performed by the microdilution technique. Antioxidant tests were performed by scavenging the DPPH and ABTS radicals, and by FRAP assay. Anti-inflammatory activity was evaluated through the inhibition of nitric oxide production in activated macrophage RAW 264.7 cells. Cell viability was assessed with an MTT assay. Results show that the flower extract revealed a powerful antimicrobial capacity against Gram-positive bacteria and strong antioxidant and anti-inflammatory activities. No significant cytotoxicity to activated macrophages was recorded. High resolution electrospray ionization mass spectrometry and nuclear magnetic resonance analysis identified two molecules with important anti-inflammatory effects: 12α-hydroxydaturametelin B and daturametelin B. Molecular docking analysis with both pro-inflammatory agents tumor necrosis factor alpha and interleukin-6 revealed that both compounds showed good binding features with the selected target proteins. Our results suggest that D. stramonium flower is a promising source of compounds with potential antioxidant, antibacterial, and anti-inflammatory activities. Isolated withanolide steroidal lactones from D. stramonium flower extract with promising anti-inflammatory activity have therapeutic potential against inflammatory disorders.

Identification of Anti-Inflammatory Compounds from Peucedanum praeruptorum Roots by Using Nitric Oxide-Producing Rat Hepatocytes Stimulated by Interleukin 1β.

The roots of Peucedanum praeruptorum Dunn and Angelica decursiva Franchet et Savatier are designated Zenko, which is a crude drug defined by the Japanese Pharmacopoeia. This crude drug is used as an antitussive and an expectorant and is included in the Kampo formula Jinsoin, which improves cough, fever, and headache. Although the anti-inflammatory effects of this crude drug have been determined, the constituents responsible for this effect remain unknown. To investigate biologically active compounds, rat hepatocytes were used, which produce proinflammatory mediator nitric oxide (NO) in response to proinflammatory cytokine interleukin 1β (IL-1β). A methanol extract of P. praeruptorum roots, which suppressed IL-1β-induced NO production, was fractionated into three crude fractions (ethyl acetate (EtOAc)-soluble, n-butanol-soluble, and water-soluble fractions) based on hydrophobicity. The EtOAc-soluble fraction markedly inhibited NO production. After this fraction was purified, three biologically active compounds were identified as praeruptorins A, B, and E, the contents of which were high. A comparison of their activities indicated that praeruptorin B exhibited the highest potency to inhibit NO production by decreasing inducible NO synthase expression and suppressed the expression of mRNAs encoding proinflammatory cytokines. Collectively, the three praeruptorins may primarily contribute to the anti-inflammatory effects of P. praeruptorum roots.

New nitric oxide production inhibitors from Syzygium bullockii.

Three undescribed triterpene glycosides syzybullosides A-C (1-3) along with fourteen known compounds were isolated from the leaves of Syzygium bullockii (Hance) Merr.& L.M. Perry, including six triterpene glycosides (1-6), four phenolics (7-9, 17), four megastigmanes (10-13), and three flavonoids (14-16). The structures of 1-17 were elucidated by extensive spectroscopic analysis, including IR, HR-ESI-MS, 1D and 2D NMR spectra. Compounds 1-10 and 12-17 inhibited nitric oxide (NO) production in lipopolysaccharide activated RAW264.7 cells with IC50 values ranging from 1.30 to 13.70µM, lower than that of the positive control compound, L-NMMA (IC50 = 33.8µM).

In vitro antiproliferative, anti-inflammatory effects and molecular docking studies of natural compounds isolated from Sarcocephalus pobeguinii (Hua ex Pobég).

Background: Sarcocephalus pobeguinii (Hua ex Pobég) is used in folk medicine to treat oxidative-stress related diseases, thereby warranting the investigation of its anticancer and anti-inflammatory properties. In our previous study, the leaf extract of S. pobeguinii induced significant cytotoxic effect against several cancerous cells with high selectivity indexes towards non-cancerous cells. Aim: The current study aims to isolate natural compounds from S. pobeguinii, and to evaluate their cytotoxicity, selectivity and anti-inflammatory effects as well as searching for potential target proteins of bioactive compounds. Methods: Natural compounds were isolated from leaf, fruit and bark extracts of S. pobeguinii and their chemical structures were elucidated using appropriate spectroscopic methods. The antiproliferative effect of isolated compounds was determined on four human cancerous cells (MCF-7, HepG2, Caco-2 and A549 cells) and non-cancerous Vero cells. Additionally, the anti-inflammatory activity of these compounds was determined by evaluating the nitric oxide (NO) production inhibitory potential and the 15-lipoxygenase (15-LOX) inhibitory activity. Furthermore, molecular docking studies were carried out on six putative target proteins found in common signaling pathways of inflammation and cancer. Results: Hederagenin (2), quinovic acid 3-O-[α-D-quinovopyranoside] (6) and quinovic acid 3-O-[β-D-quinovopyranoside] (9) exhibited significant cytotoxic effect against all cancerous cells, and they induced apoptosis in MCF-7 cells by increasing caspase-3/-7 activity. (6) showed the highest efficacy against all cancerous cells with poor selectivity (except for A549 cells) towards non-cancerous Vero cells; while (2) showed the highest selectivity warranting its potential safety as a chemotherapeutic agent. Moreover, (6) and (9) significantly inhibited NO production in LPS-stimulated RAW 264.7 cells which could mainly be attributed to their high cytotoxic effect. Besides, the mixture nauclealatifoline G and naucleofficine D (1), hederagenin (2) and chletric acid (3) were active against 15-LOX as compared to quercetin. Docking results showed that JAK2 and COX-2, with the highest binding scores, are the potential molecular targets involved in the antiproliferative and anti-inflammatory effects of bioactive compounds. Conclusion: Overall, hederagenin (2), which selectively killed cancer cells with additional anti-inflammatory effect, is the most prominent lead compound which may be further investigated as a drug candidate to tackle cancer progression.

Continuous NO dual-generation by ZnO nanoparticle conjugated with α-lipoic acid for functional biodegradable vascular stent

Coronary artery diseases are treated with a biodegradable vascular scaffold (BVS) intervention. However, BVS need to be enhanced to avoid post-insertion side effects, which can be done by controlling the nitric oxide (NO) concentration in blood vessels. NO has excellent functions, such as re-endothelialization, inhibition of smooth muscle cells, and prevention of activation and aggregation of platelets, and is essential in maintaining a healthy heart and blood vessels. In this study, a novel nanoparticle with fortified NO-generating ability was synthesized by combining two mechanisms capable of long-term NO production: the decomposition of the NO donor by the redox reaction of zinc oxide and the thiol/disulfide exchange reaction between the NO donor and α-lipoic acid. Next, a poly(L-lactic acid) (PLLA) scaffold containing synthesized nanoparticles was fabricated, and the potent functional recovery ability of unhealthy vessels was demonstrated in vitro and in vivo. The NO generated from scaffolds inhibited platelet adhesion and activation, increased endothelial cell proliferation, and decreased smooth muscle cell proliferation. In addition, the scavenging ability of reactive oxygen species and their inhibition of toxic NO production were investigated. The scaffolds implanted subcutaneously in mice suppressed the inflammatory response and induced healthy blood vessel formation, offering a new tool for designing cardiovascular stents and various biological applications.

New 5-methyl-4-hydroxycoumarin polyketide derivatives from Gerbera delavayi with anti-inflammatory activity

Five new 5-methyl-4-hydroxycoumarin polyketide derivatives (MPDs), delavayicoumarins A-E (1–5), were isolated from the whole plants of Gerbera delavayi. Among them, compounds 1–3 are the common monoterpene polyketide coumarins (MPCs), while 4 is a modified MPC with both the lactone ring contracted to a five-membered furan ring and a carboxyl at C-3, and 5 is a pair of unusual phenylpropanoid polyketide coumarin enantiomers (5a and 5b), featuring a phenylpropanoid unit at C-3. The planar structures were elucidated by spectroscopic methods and biosynthetic arguments, and the absolute configurations of 1–3, 5a and 5b were confirmed by calculated electronic circular dichroism (ECD) experiment. Furthermore, compounds 1–3, (+)-5 and (−)-5 were tested for the nitric oxide (NO) inhibitory activity by using lipopolysaccharide (LPS)-induced RAW 264.7 cells in vitro. The results showed that compounds 1–3, (+)-5 and (−)-5 remarkably inhibited NO production at the concentration of 10.0 μM, exhibiting that they have significant anti-inflammatory activity.

Analytical determination, antioxidant and anti-inflammatory activities of Bhamrung-Lohit a traditional Thai medicine.

Bhamrung-Lohit (BRL) remedy is a traditional Thai medicine (TTM). There are few reports of biological activity, the activity of its constituent plants, or quantitative analytical methods for the content of phytochemicals. In this study, we investigated antioxidant, anti-inflammatory activity, and total phenolic and flavonoid content and validated a new analytical method for BRL. Antioxidant activity was evaluated by a 2,2-diphenyl-1-picrylhydrazyl (DPPH and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical scavenging. The cellular antioxidant activity was evaluated by inhibition of the superoxide anion (O2●-) production from HL-60 cells and anti-inflammatory activity by inhibition of nitric oxide production in RAW264.7 cells. The total phenolic and flavonoid contents were analyzed using the Folin-Ciocalteu method and an aluminum chloride colorimetric assay, respectively. Validated analytical procedures were conducted according to International Conference on Harmonization (ICH) guidelines. An ethanolic extract of BRL exerted potent DPPH radical scavenging activity and moderate antioxidant and anti-inflammatory activity. Caesalpinia sappan exerted the greatest effect and the highest content of total phenolics and flavonoids. The HPLC method validated parameters that complied with ICH requirements. Each peak showed selectivity with a baseline resolution of 2.0 and precision was less than 2.0% CV. The linearity of all compounds was > 0.999 and the recovery % was within 98.0%-102.0%. The validated results demonstrated specificity/selectivity, linearity, precision, and accuracy with appropriate LOD and LOQ. BRL remedy, a TTM demonstrated antioxidant and anti-inflammatory properties. This study is the first report on the biological activity and the validation of an HPLC method for BRL remedy.

Screening of ten wild fruits extracts from Cameroon’s flora for their anti-inflammatory and antioxidant potential

Background: Wild edible fruits are good source of phenolic compounds such as anthocyanins. Those compounds are known for their anti-inflammatory potential. They interact with many mediators of the physiopathological pathway such as NF-kB which is at a central position in the inflammatory response in macrophages. This study aimed to determine the anti-inflammatory and antioxidant potential of ten wild edible fruits aqueous extracts. Methods: The antioxidant potential was evaluate through the DPPH, ABTS and OH radical scavenging activity; the ferric reducing agent power and the total antioxidant capacity assessment. Also, the total phenolic and flavonoids, anthocyanins compounds and vitamin C content were assayed. For the anti-inflammatory activity, a primary macrophage culture was used. Inflammatory mediators such as nitric oxide (NO), alkaline phosphatase, NADPH oxidase and 5-lipoxigenase activity was assayed. Results: Alchornea cordifolia and Pentaclethra macrophylla extracts showed the greatest antioxidant activity respectively (Antiradical power (AP)=5.82 mol of DPPH/mg of extract and AP=5.05 mol of DPPH/mg of extract for DPPH scavenging assay); (IC50=1.62µg/ml and IC50=1.60µg/ml for ABTS radical scavenging assay); (IC50=32.67µg/ml and IC50=70.01µg/ml for OH radical scavenging assay) and (EC50=54.06µg/ml and EC50=17.46µg/ml for ferric reducing antioxidant power) with a non-significant difference with the reference compound. For the anti-inflammatory activity, firstly it has been observed that Alchornea cordifolia’s extract exhibits the greatest potential against NADPH oxidase activity inhibition (IC50=36.20µg/ml); alkaline phosphatase activation (EC50=0.27µg/ml) with non-significant difference with the reference compound. Secondly, Pentaclethra macrophylla’s extract demonstrated a high ability to inhibit NO production (IC50=5.79µg/ml) and 5-lipoxigenase (IC50=1.41µg/ml). Conclusion: From the results obtained the assessed wild fruit extracts appears to be good sources for anti-inflammatory and antioxidant bioactive molecules. They could be beneficial for the research of new modulators for the management of inflammatory disorders.

Anti-inflammatory and Antimicrobial Potential of Three Natural Polyketides Isolated from Endophytic Fungus Phomopsis sp CAM212 against to Dysenteric Causing Pathogens

Aims: The present work aimed to evaluate the anti-amoebic, antibacterial, and anti-inflammatory potential of three natural polyketides from Phomopsis sp. CAM212.
 Study Design: Clinical isolates of E.histolytica, E.coli ATCC25922 strain, primary peritoneal mouse macrophages and three polyketides were used.
 Places and Duration of Study: Laboratory of Pharmacology and Toxicology, Laboratory of Medical Microbiology, Faculty of Science, University of Yaounde 1 between May and December 2022.
 Methodology: During this work, we evaluated the ability of three natural polyketides from Phomopsis sp to inhibit the growth of germs responsible for amoebic and bacillary dysentery. First, the anti-amoebic activity was carried out on clinical isolates of E. histolytica in polyxenic culture. Subsequently, we evaluated the antibacterial potential on a strain of E. coli ATCC25922. Finally, the anti-inflammatory potentials were evaluated on a primary culture of SC activated macrophages through inhibition of nitric oxide (NO) production, activation of phosphatase alcaline (ALP) and inhibition of 5-lipoxygenase (5-LOX).
 Results: It emerges from this work that among compounds, phomopsinin B, presented the highest anti-amoebic potential (84.4 % inhibition after 72h) and the highest antibacterial potential (MIC=12.5µg/mL and MBC/MIC=2). Phomopsini A and phomopsini A acetate showed moderate anti amoebic and antibacterial potentials. However, all these activities remain lower than that of metronidazole and ciprofloxacin (90% of amoebic inhibition after 72h; MIC=0.72µg/mL and MBC/MIC=4). Subsequently, all tested compounds were nontoxic on primary macrophages. Phomopsinin B exhibited a great anti-inflammatory potential through the inhibition of NO production (IC50=1.72±0.91µg/mL); inihibition of 5-LOX activity (IC50=36.97±7.12µg/mL) and activation of ALP activity (IC50=0.13±0.01µg/mL) as compared to Baicalin the standard. The anti-inflammatory potential of phomopsinin A and phomopsinin A acetate were lower compared to baicalin.
 Conclusion: Ultimately, among compounds tested, phomopsinin B exhibited the best anti-amoebic, antibacterial and ant-inflammatory potential similar to the respective standards within the limits of the tests carried out.