Anti-Acne Vulgaris Potential of the Ethanolic Extract of Mesua ferrea L. Flowers

Acne vulgaris is a common chronic inflammatory skin disease. In the present study, we reported the anti-acne vulgaris effect of the Mesua ferrea (M. ferrea) flower extract. The extract was evaluated for three anti-acne-causing bacteria properties including Cutibacterium acnes (C. acnes), Staphylococcus epidermidis (S. epidermidis) and Staphylococcus aureus (S. aureus). The results indicated that the M. ferrea flower extract could be considered as the bactericidal agent against S. epidermidis and S. aureus with MIC values of 0.78 and 6.25 mg mL−1 and MBC values of 1.56 and 12.50 mg mL−1 and the bacteriostatic agent against C. acnes with MIC and MBC values of 3.12 and 25.00 mg mL−1, respectively. The extract at a concentration of 25 µg mL−1 also presented potent anti-inflammatory activity with a significant decrease of nitric oxide (NO) and tumor necrosis factor (TNF)-α productions in RAW 264.7 macrophage cells stimulated by LPS. In addition, the extract showed moderate to weak anti-oxidative capacities against DPPH, ABTS, FRAP and NO assays and also showed weak anti-tyrosinase activity. M. ferrea flower extract may serve as the alternative natural anti-acne formulations.

Immune Responses Are Differentially Regulated by Root, Stem, Leaf, and Flower Extracts of Female and Male CBD Hemp (Cannabis sativa L.) Plants

Industrial hemp (Cannabis sativa L.) has many applications, including the production of textiles, agricultural extracts, nutritional products, and botanicals enriched with cannabinoids and full-spectrum terpenes naturally present in the plant. In this study, the dynamics of distribution and accumulation of 10 main cannabinoids in hemp were quantified. Hemp bioactive compounds were evaluated for anti-inflammatory activity in lipopolysaccharide-induced RAW 264.7 macrophage cells. While all tissues of hemp showed moderate anti-inflammatory properties, female flowers demonstrated the highest activity. CBD showed the strongest anti-inflammatory activity with suppression of nitric oxide production at 2 μg/mL and the reduced expressions of the pro-inflammatory genes COX-2, IL-6, and TNF-α at as low as 2 ng/mL. The topical hemp inflorescences (1–50 μg/mL) and CBD alone (20–200 ng/mL) also improved mitochondrial respiration. These data contribute to the future development of agricultural and plant management techniques to produce hemp with specific metabolite profiles to selectively support immune health.

Thymol and Piperine-Loaded Poly(D,L-lactic-co-glycolic acid) Nanoparticles Modulate Inflammatory Mediators and Apoptosis in Murine Macrophages

This study aimed at preparing and characterizing thymol, eugenol, and piperine-loaded poly(D,L-lactic-co-glycolic acid) nanoparticles and evaluating the effect on inflammatory mediators secretion and apoptosis in Raw 264.7 macrophage cells. Nanoparticles were produced by the solvent evaporation technique. Dynamic light scattering and scanning electron microscopy were used to study the physicochemical characteristics. Raw 264.7 macrophage cells were used as a model for in vitro assays. The 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium assay was used to determine the cytotoxicity of the formulated nanoparticles. An annexin V apoptosis detection kit was used to assess apoptosis. Nitric oxide production was determined using the Griess reagent, and the inflammatory mediators level was evaluated with Th1/Th2 cytokine and fluorometric cyclooxygenase kits. The loaded nanoparticles showed a particle size around 190 nm with a low polydispersity between 0.069 and 0.104 and a zeta potential between–1.2 and–9.5 mV. Reduced cytotoxicity of nanoparticles compared to free molecules against Raw 264.7 macrophage cells was observed and seemed to occur through a mechanism associated with apoptosis. A decrease in cyclooxygenase enzyme activity with an increasing concentration was observed. Both free molecules and nanoparticles showed their capacity to modulate the inflammatory process mostly by inhibiting the investigated inflammatory cytokines. The data presented in this study indicate that thymol and piperine-loaded poly(D,L-lactic-co-glycolic acid nanoparticles could serve as a novel anti-inflammatory colloidal drug delivery system with reduced toxicity. However, further study should be considered to optimize the formulation’s loading capacity and thereby probably enhance their bioactivity in treating inflammatory diseases.

The Labile Iron Pool Reacts Rapidly and Catalytically with Peroxynitrite

While investigating peroxynitrite-dependent oxidation in murine RAW 264.7 macrophage cells, we observed that removal of the Labile Iron Pool (LIP) by chelation increases the intracellular oxidation of the fluorescent indicator H2DCF, so we concluded that the LIP reacts with peroxynitrite and decreases the yield of peroxynitrite-derived oxidants. This was a paradigm-shifting finding in LIP biochemistry and raised many questions. In this follow-up study, we address fundamental properties of the interaction between the LIP and peroxynitrite by using the same cellular model and fluorescence methodology. We have identified that the reaction between the LIP and peroxynitrite has catalytic characteristics, and we have estimated that the rate constant of the reaction is in the range of 106 to 107 M−1s−1. Together, these observations suggest that the LIP represents a constitutive peroxynitrite reductase system in RAW 264.7 cells.

In vitro Cytotoxicity and Antioxidant Study of Rhodomyrtus tomentosa (Aiton) Hassk. Ethanolic Leaf Extract on LPS-induced RAW 264.7 Macrophage Cells

Aims: Ethanol extract of Rhodomyrtus tomentosa leaves found specifically on Malaysian soil was used to further investigate the antioxidant properties and cytotoxicity against RAW 264.7 macrophage cells in the search for a safer and effective natural antioxidant agent. Study Design: Antioxidant potential of R. tomentosa were analyzed through series of spectrometric assays and cell-based bioassays model. Place and Duration of Study: This study was carried out at Faculty of Pharmacy, Universiti Teknologi MARA (UiTM), Puncak Alam Campus, 43200, Selangor, Malaysia from the year of 2019 to 2021. Methodology: R. tomentosa leaves were subjected to extraction with 95% ethanol. The extracts were then denoted as ethanolic leaves extract of R. tomentosa or EtRT extract. EtRT extract were then screen for its antioxidant activity (AOA) and total antioxidant capacity (TAC) through DPPH radical scavenging assay and ferric reducing antioxidant power (FRAP) assay. After that, EtRT extract were brought to observe its toxicity against RAW 264.7 macrophage cells in MTT assay. Once their toxicity was obtained, EtRT extracts were finally tested for their ability to inhibit intracellular reactive oxygen species (ROS) and nitric oxide (NO) inhibition in RAW 264.7 macrophage cells to further analyze their antioxidant properties. Results: In this study, EtRT extracts dose dependently showed the ability to scavenge DPPH radicals and reduce ferric ions during DPPH radicals scavenging activity assay and ferric reducing antioxidant power assay (FRAP), respectively. In DPPH radical scavenging activity assay, EtRT extracts showed EC50 value at 12.37 ± 1.73 µg/mL with ARP value of 0.08 almost as near as ascorbic acid’s ARP value which is 0.09. Further into the study, EtRT extract were not cytotoxic to RAW 264.7 macrophage cells at concentrations 3.91 µg/mL and lower which showed more than 86.4% cell viability with IC50 value at 204.70 ± 5.30 µg/mL. EtRT extract possessed the ability to inhibit ROS production on LPS-induced RAW 264.7 macrophage cells at 7.813 µg/mL and lower, with the highest concentration can reduce up to 30.20% ± 1.01 out of the total ROS produced by the induced cells. Furthermore, EtRT extract also have evidenced that it is able to significantly inhibit NO production by the LPS-induced RAW 264.7 macrophage cells at 7 µg/mL and lower being the highest at 56.73% ± 0.11 inhibition of the highest concentration tested. Conclusions: This study suggests that EtRT extracts have the potential to reduce LPS-induced oxidative stress due to the antioxidant activities of phenolic compounds in the extracts, and that at low doses, EtRT extracts had low to no cytotoxicity on RAW 264.7 macrophage cells. As a result, EtRT extract could be a promising natural medicinal agent for the treatment of oxidative stress.

Antifungal Activity of Isolated Compounds from the Leaves of Combretum erythrophyllum (Burch.) Sond. and Withania somnifera (L.) Dunal against Fusarium Pathogens

Crop diseases caused by Fusarium pathogens, among other microorganisms, threaten crop production in both commercial and smallholder farming. There are increasing concerns about the use of conventional synthetic fungicides due to fungal resistance and the associated negative effects of these chemicals on human health, livestock and the environment. This leads to the search for alternative fungicides from nature, especially from plants. The objectives of this study were to characterize isolated compounds from Combretum erythrophyllum (Burch.) Sond. and Withania somnifera (L.) Dunal leaf extracts, evaluate their antifungal activity against Fusarium pathogens, their phytotoxicity on maize seed germination and their cytotoxicity effect on Raw 264.7 macrophage cells. The investigation led to the isolation of antifungal compounds characterized as 5-hydroxy-7,4′-dimethoxyflavone, maslinic acid (21-hydroxy-3-oxo-olean-12-en-28-oic acid) and withaferin A (4β,27-dihydroxy-1-oxo-5β,6β-epoxywitha-2-24-dienolide). The structural elucidation of the isolated compounds was established using nuclear magnetic resonance (NMR) spectroscopy, mass spectroscopy (MS) and, in comparison, with the available published data. These compounds showed good antifungal activity with minimum inhibitory concentrations (MIC) less than 1.0 mg/mL against one or more of the tested Fusarium pathogens (F. oxysporum, F. verticilloides, F. subglutinans, F. proliferatum, F. solani, F. graminearum, F. chlamydosporum and F. semitectum). The findings from this study indicate that medicinal plants are a good source of natural antifungals. Furthermore, the isolated antifungal compounds did not show any phytotoxic effects on maize seed germination. The toxicity of the compounds A (5-hydroxy-7,4′-dimethoxyflavone) and AI (4β,27-dihydroxy-1-oxo-5β,6β-epoxywitha-2-24-dienolide) was dose-dependent, while compound B (21-hydroxy-3-oxo-olean-12-en-28-oic acid) showed no toxicity effect against Raw 264.7 macrophage cells.