The Clock-modulatory Activity of Nobiletin Suppresses Adipogenesis Via Wnt Signaling

Abstract

The circadian clock machinery exerts transcriptional control to modulate adipogenesis and its disruption leads to the development of obesity. Here we report that Nobiletin, a circadian clock amplitude-enhancing molecule, displays anti-adipogenic properties via activation of Wnt signaling pathway that is dependent on its clock modulation. Nobiletin augmented clock oscillatory amplitude with period lengthening in the adipogenic mesenchymal precursor cells and preadipocytes, accompanied by an induction of Bmal1 and clock components within the negative feedback arm. Consistent with its clock-modulatory activity, Nobiletin strongly inhibited the lineage commitment and terminal differentiation of adipogenic progenitors. Mechanistically, we show that Nobiletin induced the re-activation of Wnt signaling during adipogenesis via transcriptional up-regulation of key components within this pathway. Furthermore, Nobiletin administration in mice markedly reduced adipocyte hypertrophy, leading to a significant loss of fat mass and reduction of body weight. Lastly, Nobiletin inhibited the differentiation of primary preadipocytes and this effect was dependent on a functional clock regulation. Collectively, our findings uncover a novel activity of Nobiletin in suppressing adipocyte development in a clock-dependent manner, implicating its potential application in countering obesity and associated metabolic consequences.

Anti-adipogenic properties of clock activator chlorhexidine and a new derivative

“…0.25% Trypsin was used for digestion and subculture of these cell lines. For adipogenic differentiation, induction media containing 1.6μM insulin, 1μM dexamethasone, 0.5mM IBMX and 0.5 uM Rosiglitazone was used for 3 days followed by maintenance medium with insulin for 3 days for 3T3-L1, and for 5 days for C3H10T1/2 cells, as previously described (17,27).…”

“…preadipocytes and C3H10T1/2 mesenchymal stem cells obtained from ATCC were used for Per2::dLuc lentiviral transduction and stable clone selection using puromycin, as described previously (17,28). Briefly, cells were transfected with lentiviral packaging plasmids (pSPAX.2 and pMD2.G) and lentivirus vectors Per2::dLuc using PEI Max (Polysciences).…”

“…Notably, activation of the clock repressor Rev-erbα by specific agonists, a CLOCK/Bmal1 direct target gene, demonstrated anti-obesity efficacy in mice with improvement of dyslipidemia (16). On the other hand, RORα modulation by a natural ligand, Nobiletin, inhibited adipogenesis 4 with in vivo effects on countering obesity (17). Thus, targeting the circadian clock function in modulating adipocyte development may provide novel pharmacological interventions for obesity and related metabolic consequences.…”

The Interconnection between Hepatic Insulin Resistance and Metabolic Dysfunction-Associated Steatotic Liver Disease—The Transition from an Adipocentric to Liver-Centric Approach

“…The modulation of IRE1 activity is suggested as a potential therapeutic target, since IRE1 kinase inhibitor application significantly attenuated nucleotide-binding oligomerization domain (NOD) 1 and NOD2-mediated proinflammatory response [45]. Novel therapeutic strategies include targeting clock genes and the resynchronization of disrupted biological clocks, which are involved in metabolic derangement [155,156]. Recent studies on suggested potential therapeutic agents are shown in the table, which can be further investigated for more beneficial effects and detailed mechanisms of action (Table 1).…”

“…Decrease in lipogenesis markers (SREBP1 and PPARγ) [157] Andrographis paniculata (Burm. f.) Nees Modulation of the IRS-1/GLUT-2 pathway due to IL-6 inhibition [158] Icariin Reduction in thioredoxin-interacting protein (TXNIP) and suppression of ER stress [159] Lunasin Regulation of anti-inflammation, anti-oxidation, and glucose utilization and amelioration of glucose uptake [160] Glucosamine Inhibition of the LPS/TLR4/NF-κB pathway [161] D-allulose Anti-inflammatory effects through the suppression of INF-γ and the enhancement of macrophage function [162] Syzygium cumini Partial agonism of PPARγ and an increase in the expression of adiponectin, an insulin sensitizer [163] Morin and 1-deoxynojirimycin Suppression of cytokine signaling 3 and CD36/Serbp1/Fas signaling and promotion of PPARγ [164] Nobiletin Suppression of adipocyte development in a clock-dependent manner [155]…”