Andrographolide (AG) has been shown to have several medicinal and pharmaceutical effects, such as antimicrobial, anti-inflammatory, antioxidant, anti-diabetic, and anti-malarial activities. Moreover, studies to assess the pharmacological effect of AG on the metabolic changes of uninfected red blood cells (uRBCs) have not yet been investigated. This study aims to evaluate the pharmacological effects of AG compared to chloroquine (CQ) on the metabolic variations of uRBCs in vitro using a proton nuclear magnetic resonance (1H-NMR)-based metabolomics approach coupled with multivariate data analysis (MVDA). Forty-one metabolites were successfully identified by 1H-NMR. The results of the unsupervised data analysis principal component analysis (PCA) showed ideal differentiation between AG and CQ. PC1 and PC2 accounted for 71.4% and 17.7% of the explained variation, respectively, with a total variance of 89.10%. Based on S-plot and VIP values, a total of 28 and 32 metabolites were identified as biomarkers in uRBCs-AG and uRBCs-CQ, respectively. In uRBCs treated with AG, ten metabolic pathways were determined to be disturbed, including riboflavin metabolism, d-glutamate and d-glutamine metabolism, phenylalanine metabolism, glutathione metabolism, proline and arginine metabolism, arginine biosynthesis, citrate cycle, glycolysis/gluconeogenesis, and pyruvate metabolism as well as alanine, aspartate, and glutamate metabolism. In contrast, in CQ-treated uRBCs, nine affected metabolic pathways were determined, which involved the same metabolic pathways for uRBCs-AG, except for glutathione metabolism. These findings suggest an evident relationship between AG and CQ associated with metabolic changes in intact RBCs after being exposed to the treatment. The metabolomics results could allow useful comprehensive insights into the underlying mechanism of the action of AG and CQ on red blood cells. Consequently, the 1H-NMR-based metabolomics approach was successfully utilized to identify the pharmacological effects of AG and CQ on the metabolic variations of uRBCs.

Arbutus pavarii Pamp is a medicinal plant commonly used by local tribes in East Libya

for the treatment of many diseases, such as gastritis, renal infections, cancer and kidney diseases.

In this study, the antibacterial activity of the leaf and stem bark extracts of the plant against

methicillin-resistant Staphylococcus aureus (MRSA), as well as the metabolite profiles of the bioactive

fractions, was investigated. The antibacterial activity was determined by disc diffusion method,

minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), while the

microbial reduction by the bioactive fraction was evaluated using time–kill test. The bioactive

fraction was further subjected to ultrahigh-performance liquid chromatography–mass spectrometry

(UHPLC-ESI-MS/MS) analysis to putatively identify the chemical constituents contained therein.

All the extracts and fractions showed different levels of antibacterial activity on the tested MRSA

strains. The highest total antibacterial activity, i.e., 4007.6 mL/g, was exhibited by the crude leaf

methanolic extract. However, the ethyl acetate fraction of the leaf showed moderate to significant

antibacterial activity against MRSA at low MIC (0.08–1.25 mg/mL). Metabolite profiling of this fraction

using UHPLC-ESI-MS/MS resulted in the putative identification of 28 compounds, which included

phenolic acids, flavan-3-ols and flavonols. The results of this study showed that the ethyl acetate

fraction of Arbutus pavarii leaf possessed potential antibacterial activity against MRSA and hence can

be further explored for pharmaceutical applications as a natural antibacterial agent.

0

0