Biflavonoid Methylchamaejasmin and Khaya grandifoliola Extract Inhibit NLRP3 Inflammasome in THP-1 Cell Model of Neuroinflammation.

Neuroinflammation is a common hallmark of Alzheimer's disease (AD), with NLRP3 inflammasome proven to be activated in microglia of AD patients' brains. In this study, a newly isolated biflavonoid (7,7'-di-O-methylchamaejasmin/M8) and a crude extract of the plant Khaya grandifoliola (KG) were investigated for their inhibitory effect on inflammasome activation. In preliminary experiments, M8 and KG showed no cytotoxicity on human macrophage-like differentiated THP-1 cells and exhibited anti-inflammatory inhibition of nitric oxide produced following lipopolysaccharide stimulation. Furthermore, M8 and KG blocked IL-1ß and IL-18 production by reducing NLRP3 inflammasome components including NFκB, NLRP3, Caspase-1, pro-IL-1ß, and pro-IL-18 at the mRNA and protein levels. Regarding the formation of ASC (apoptosis-associated speck-like protein containing a CARD) specks during inflammasome activation, the size and fluorescent intensity of the existing specks were unchanged across all treatment conditions. However, M8 and KG treatments were shown to prevent further speck formation. In addition, experiments on amyloid ß phagocytosis showed that M8 and KG pretreatments can restore the phagocytic activity of THP-1 cells, which was impaired following inflammasome activation. Altogether, our findings describe for the first time a promising role of biflavonoids and KG extract in preventing inflammasome activation and protecting against neuroinflammation, a key factor in AD development.

Phytochemical Study of Aqueous Extract of Ochna schweinfurthiana F. Hoffm Powder Bark and Evaluation of Their Anti-Inflammatory, Cytotoxic, and Genotoxic Properties.

Ochna schweinfurthiana has been used in traditional medicine to treat pain, inflammation, and arthritis. It is a rich source of complex dimers of flavonoids with potential use as templates for the development of therapeutic drugs. Hence, the aim of this study was to study the phytochemical content and evaluate the in vitro cytotoxic, genotoxic, and anti-inflammatory activities of the aqueous extract of Ochna schweinfurthiana bark (OSE). Phytochemical study was carried out according to LC-MS procedures, while isolation was carried out using thin layer and column chromatographies. Cytotoxicity was investigated by the mitochondrial viability [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT) method while genotoxicity potential of the extract was ascertained using the Salmonella typhimurium test strains TA98 and TA100. The anti-inflammatory effect of OSE was evaluated by the in vitro inhibition of 15-lipooxygenase enzyme and bovine serum albumin denaturation (BSA) assays. The investigation of compounds extracted from OSE led to the identification and isolation of six known compounds, namely, hemerocallone (9), 6,7-dimethoxy-3'-4'-dimethoxyisoflavone (10), lithospermoside (13), amentoflavone (14), agathisflavone (15), and ß-D-fructofuranosyl-α-D-glucopyranoside (17). In the anti-inflammatory assay, aqueous extracts of the bark showed selective inhibition of 15-lipooxygenase with IC50 value of 32.2 ± 0.36 µg/mL and the result of the bovine serum albumin denaturation assay with IC50 value of 130± 5.78 µg/mL showed moderate activity. The toxicity assay indicated that OSE are noncytotoxic on Vero cell line with LC50 value of 50 mg/mL and nongenotoxic toward Salmonella typhimurium tester strain TA98 and TA100. Result from this study supports the traditional use of the selected medicinal plants in Cameroon for the treatment of inflammatory conditions. Noncytotoxicity and nongenotoxicity of OSE suggest that this plant is safe for use.

Cytotoxic flavonoids and other constituents from the stem bark of Ochna schweinfurthiana.

Seven flavonoids, hemerocallone (1), 6,7-dimethoxy-3',4'-dimethoxyisoflavone (2), amentoflavone (4), agathisflavone (6), cupressuflavone (8), robustaflavone (9) and epicatechin (10), together with three other compounds, lithospermoside (3), ß-D-fructofuranosyl-α-D-glucopyranoside (5) and 3ß-O-D-glucopyranosyl-ß-stigmasterol (7), were isolated from the ethyl acetate extract of the stem bark of Ochna schweinfurthiana F. Hoffm. All the compounds were characterised by spectroscopic and mass spectrometric methods, and by comparison with literature data. Cytotoxicity of the extracts and compounds against cervical adenocarcinoma (HeLa) cells was evaluated by MTT assay. Compounds 4 and 6 exhibited good cytotoxic activity, with IC50 values of 20.7 and 10.0 µM, respectively.