Effect of a methanolic extract of Salvadora oleoides Decne. on LPS-activated J774 macrophages, its in vitro and in vivo toxicity study and dereplication of its chemical constituents. (2022)

• Record Type: Journal Article
• Authors: Khaliq, Hafiz Abdul; Ortiz, Sergio; Alhouayek, Mireille; Neyts, Tanguy; Muccioli, Giulio G.; Quetin-Leclercq, Joëlle
• Is Part Of: Toxicology reports. Volume 9(2022)
• Page Start: 1742
• Page End: 1753
• Publication Date: 2022
• Subjects: OECD -- Oral toxicity -- Inflammation -- GNPS -- Flavonoids; OLM Salvadora oleoides methanolic extract; Toxicology -- Periodicals; Clinical toxicology -- Periodicals; Drug-Related Side Effects and Adverse Reactions; Hazardous Substances; Poisoning; Toxicology; Electronic journals; Periodicals; Periodicals; 571.9505
• DOI: 10.1016/j.toxrep.2022.09.004
• Languages: English
• ISSNs: 2214-7500

Abstract

Abstract: Salvadora oleoides is used in Pakistani traditional medicine to treat inflammatory conditions, piles, boils, and ulcers. To evaluate the anti-inflammatory potential of S. oleoides (a mixture of aerial branches, leaves, and stem bark), we prepared crude extracts in Soxhlet apparatus by successively using different solvents and found the methanolic extract (OLM) to significantly inhibit the LPS-induced expression of pro-inflammatory cytokines and enzymes in J774 macrophages, at 50 µg/mL concentration. We also analysed the chemical constituents of OLM by dereplication, performed by HPLC-MS/MS and molecular networking. The major detected constituents were flavonoids and phenolic acids glycosides, most of them identified for the first time in S. oleoides . We also evaluated the toxicity of OLM against five cell lines, namely Caco-2, HepG2, HeLa, J774, and WI-38 by MTT assay. The IC50 was found to be higher than 100 µg/mL against these five cell lines after 72 h treatment. Furthermore, OLM was tested in mice for acute and sub-acute oral toxicity according to the guidelines of the Organisation for Economic Co-operation and Development (OECD). OLM was found non-toxic, except for some fibrosis observed in the spleens of treated mice in the sub-acute oral toxicity test. Our results confirm the anti-inflammatory potential of OLM and that it could be tested in in vivo inflammatory models, but its effect on the spleen should be considered before designing the experiments. Graphical Abstract: ga1 Highlights: MeOH extract of S. oleoides inhibited LPS-induced expression of pro-inflammatory cytokines and enzymes in J774 macrophages. Major chemical constituents of OLM were putatively identified by dereplication, using HPLC-MS/MS and molecular networking. IC50 value of OLM was > 100 µg/mL when tested by MTT assay against Caco-2, HepG2, HeLa, J774, and WI-38 cells. OLM was found non-toxic in mice when tested for acute oral toxicity, according to the guidelines of OECD. In sub-acute oral toxicity study, fibrosis was observed in the spleens of the treated mice.