Cimicifuga racemosa extracts (CREs) have gained well-established use for the treatment of menopausal symptoms such as hot flushes and excessive sweating, and weight gain. While the clinical effects of CREs have been well documented, the mechanisms underlying these effects are largely unknown. More recently, the metabolic effects of the CRE Ze 450 were demonstrated in cultured cells in vitro and in mouse models of obesity in vivo. At the molecular level, metabolic regulation, enhanced insulin sensitivity, and increased glucose uptake were linked to the activation of AMP-activated protein kinase (AMPK). Therefore, we tested the effects of Ze 450 on AMPK phosphorylation and thus activation in cells from different tissues, i.e., murine C2C12 myoblast cells, human HEPG2 liver cells, mouse HT22 neuronal cells, and in murine 3T3L1 adipocytes. Using a FRET-based HTRF-assay, we found that Ze 450 induced AMPK phosphorylation and the activation of this key enzyme of metabolic regulation in cells from various different tissues including C2C12 (muscle), HEPG2 (liver), HT22 (hippocampal), and 3T3-L1 (adipocyte) cells. In C2C12 muscle cells, enhanced AMPK activation was accompanied by reduced mitochondrial respiration and enhanced glucose uptake. Further, Ze 450 enhanced the resilience of the cells against oxidative death induced by ferroptosis inducers erastin or RSL3. Our findings suggest a general effect of Cimicifuga racemosa on AMPK activation in different tissues and across species. This may have a significant impact on expanded therapeutic applications of Ze 450, since AMPK activation and the related metabolic effects have been previously associated with anti-aging effects and the prevention of the metabolic syndrome.

Chronic inflammation is a key contributor to the pathogenesis of numerous diseases, including cardiovascular, metabolic, and neurodegenerative disorders. Nutraceutical strategies targeting inflammatory pathways are of increasing interest, particularly those based on omega-3 fatty acids. The objective of this study was to evaluate the anti-inflammatory effects of two omega-3-based nutraceutical formulations, Omega 3 Plus and Omega 3 Orange, in primary human monocytes. Primary human monocytes were isolated from peripheral blood of a healthy donor and cultured under standardized conditions. Cells were pre-treated with different concentrations of the test formulations and subsequently stimulated with lipopolysaccharide (LPS, 10 ng/mL) for 24 h. Cell viability was assessed using the AlamarBlue assay. The release of pro-inflammatory mediators, including TNF-α, IL-1β, IL-6, MCP-1, IL-8, and prostaglandin E2 (PGE2), as well as the anti-inflammatory cytokine IL-10, was quantified using ELISA. Both formulations were well tolerated at concentrations up to 2.5%, with no significant cytotoxic effects. LPS stimulation induced a robust increase in inflammatory mediator release. Pre-treatment with Omega 3 Plus and Omega 3 Orange resulted in a significant, dose-dependent inhibition of pro-inflammatory cytokines, including TNF-α, IL-1β, and IL-6 (up to ~70% reduction). MCP-1 was moderately reduced, whereas IL-8 was only minimally affected. Notably, Omega 3 Orange exhibited a pronounced inhibition of PGE2 production (up to ~95%), while Omega 3 Plus reduced PGE2 levels by approximately 80%. Neither formulation induced IL-10 production in unstimulated cells. These findings demonstrate that both omega-3-based nutraceutical formulations exert potent anti-inflammatory effects in primary human monocytes, primarily through the inhibition of pro-inflammatory cytokines and PGE2. The strong suppression of PGE2 is consistent with a possible modulation of pathways involved in prostaglandin synthesis. These results support the potential application of such formulations in inflammation-associated conditions and warrant further mechanistic and clinical investigation.