Background: Yeast Pah1p phosphatidate phosphatase dephosphorylates phosphatidate to generate diacylglycerol for lipid synthesis. Results: Pah1p was phosphorylated by the Pho85p-Pho80p protein kinase-cyclin complex on seven sites contained within a (Ser/Thr)-Pro motif. Conclusion:The phosphorylation inhibited Pah1p activity, its interaction with the membrane, and triacylglycerol synthesis. Significance: Pho85p-Pho80p plays a role in lipid metabolism through its phosphorylation and regulation of Pah1p.

Nitrogen mustard (NM) is an alkylating agent known to cause extensive pulmonary injury progressing to fibrosis. This is accompanied by a persistent macrophage inflammatory response. In these studies, we characterized the phenotype of macrophages accumulating in the lung over time following NM exposure. Treatment of rats with NM (0.125 mg/kg, intratracheally) resulted in an increase in CD11b 1 macrophages in histologic sections. These cells consisted of inducible nitric oxide synthase 1 (iNOS) proinflammatory M1 macrophages, and CD68 1 M1 macrophages and mature CD43 2 M2 macrophages, which increased sequentially. Time-related increases in M1 (iNOS, IL-12a, COX-2, TNF-a, matrix metalloproteinase-9, matrix metalloproteinase-10) and M2 (IL-10, pentraxin-2, connective tissue growth factor, ApoE) genes, as well as chemokines/ chemokine receptors associated with trafficking of M1 (CCR2, CCR5, CCL2, CCL5) and M2 (CX 3 CR1, fractalkine) macrophages to sites of injury, were also noted in macrophages isolated from the lung after NM. The appearance of M1 and M2 macrophages in the lung correlated with NM-induced acute injury and the development of fibrosis, suggesting a potential role of these macrophage subpopulations in the pathogenic response to NM.

Kaempferol is a flavonoid present in Kaempferia galanga and Opuntia ficus indica var. saboten. Recent studies have suggested that it has anti-oxidant, anti-inflammatory, anti-cancer, and anti-obesity effects. In this study, we focused on the anti-adipogenic effects of kaempferol during adipocyte differentiation. The results showed that kaempferol inhibits lipid accumulation in adipocytes and zebrafish. Oil Red O and Nile Red staining showed that the number of intracellular lipid droplets decreased in adipocytes and zebrafish treated with kaempferol. LPAATθ (lysophosphatidic acid acyltransferase), lipin1, and DGAT1 (triglyceride synthetic enzymes) and FASN and SREBP-1C (fatty acid synthetic proteins) showed decreased expression levels in the presence of kaempferol. In addition, treatment of kaempferol showed an inhibitory activity on cell cycle progression. Kaempferol delayed cell cycle progression from the S to G2/M phase through the regulation of cyclins in a dose-dependent manner. Kaempferol blocked the phosphorylation of AKT (protein kinase B) and mammalian target of rapamycin (mTOR) signaling pathway during the early stages of adipogenesis. In addition, kaempferol down-regulated pro-early adipogenic factors such as CCAAT-enhancer binding proteins β (C/EBPβ), and Krüppel-like factors (KLFs) 4 and 5, while anti-early adipogenic factors, such as KLF2 and pref-1(preadipocyte factor-1), were upregulated. These kaempferol-mediated regulations of early adipogenic factors resulted in the attenuation of late adipogenic factors such as C/EBPα and peroxisome proliferator-activated receptor γ (PPARγ). These results were supported in zebrafish based on the decrease in lipid accumulation and expression of adipogenic factors. Our results indicated that kaempferol might have an anti-obesity effect by regulating lipid metabolism.

Background:In yeast, phospholipid synthesis is regulated by zinc deficiency. Results: PAH1-encoded phosphatidate phosphatase activity was induced in zinc-deficient cells by a transcriptional mechanism. Conclusion: The zinc-mediated regulation of phosphatidate phosphatase affects phospholipid synthesis by controlling phosphatidate and diacylglycerol. Significance: The transcriptional regulation of phosphatidate phosphatase plays an important role in controlling phospholipid synthesis by zinc.

Inflammatory macrophages have been implicated in hepatotoxicity induced by the analgesic, acetaminophen (APAP). In these studies we characterized the phenotype of macrophages accumulating in the liver following APAP intoxication and evaluated the role of galectin-3 (Gal-3) in macrophage activation. Administration of APAP (300 mg/kg, i.p.) to wild type mice resulted in the appearance of two distinct subpopulations of CD11b+ cells in the liver, which expressed high or low levels of the monocyte/macrophage activation marker Ly6C. Whereas CD11b+/Ly6Chi macrophages exhibited a classically activated proinflammatory phenotype characterized by increased expression of TNF-α, inducible nitric oxide synthase (iNOS), and CCR2, CD11b+/Ly6Clo macrophages were alternatively activated, expressing high levels of the anti-inflammatory cytokine, IL-10. APAP intoxication was also associated with an accumulation of Gal-3+ macrophages in the liver; the majority of these cells were Ly6Chi. APAP-induced increases in CD11b+/Ly6Chi macrophages were significantly reduced in Gal-3−/− mice. This was evident 72 h post-APAP and was correlated with reduced expression of the classical macrophage activation markers, iNOS, IL-12, and TNF-α, as well as the proinflammatory chemokines, CCL2 and CCL3, and chemokine receptors CCR1, and CCR2. Conversely, numbers of CD11b+/Ly6Clo macrophages increased in livers of APAP-treated Gal-3−/− mice. This was associated with increased expression of the alternative macrophage activation markers Ym1 and Fizz1, increased liver repair and reduced hepatotoxicity. These data demonstrate that both classically and alternatively activated macrophages accumulate in the liver following APAP intoxication; moreover, Gal-3 plays a role in promoting a persistent proinflammatory macrophage phenotype.

Oleuropein is one of the primary phenolic compounds present in olive leaf. In this study, the anti-inflammatory effect of oleuropein was investigated using lipopolysaccharide (LPS)-stimulated RAW 264.7 and a zebrafish model. The inhibitory effect of oleuropein on LPS-induced NO production in macrophages was supported by the suppression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). In addition, our enzyme immunoassay showed that oleuropein suppressed the release of pro-inflammatory cytokines such as interleukin-1β (IL-1β) and interleukin-6 (IL-6). Oleuropein inhibited the translocation of p65 by suppressing phosphorylation of inhibitory kappa B-α (IκB-α). Oleuropein also decreased activation of ERK1/2 and JNK, which are associated with LPS-induced inflammation, and its downstream gene of AP-1. Furthermore, oleuropein inhibited LPS-stimulated NO generation in a zebrafish model. Taken together, our results demonstrated that oleuropein could reduce inflammatory responses by inhibiting TLR and MAPK signaling, and may be used as an anti-inflammatory agent.

Our data demonstrate that dieckol inhibits lipid accumulation via activation of AMPKα signaling and cell-cycle arrest.

Abstract:Wild herbs, which are edible plants that grow in mountainous areas, have diverse biological effects such as anti-obesity and anti-cancer activities. The aim of this study was to evaluate the total phenolic and flavonoid contents as well as the antioxidant activity of methanol extracts of Aster scaber, Ligularia fischeri, Kalopanax pictus, Codonopsis lanceolata, and Cirsium setidens and to assess their effects on lipid accumulation and reactive oxygen species (ROS) production during adipogenesis of 3T3-L1 cells. The results revealed that among the five studied wild herb extracts, Ligularia fischeri showed the highest total phenolic contents (215.8 ± 14.2 mg GAE/g) and Aster scaber showed the highest total flavonoid content (103.9 ± 3.4 mg RE/g). Furthermore, Aster scaber and Ligularia fischeri extracts showed higher antioxidant activity than the other wild herbs. Regarding anti-adipogenic activity, the Cirsium setidens extract significantly inhibited lipid accumulation (~80%) and ROS production (~50%) during adipogenesis of 3T3-L1 cells compared with control cells. These results suggest that wild herbs could be used for the development of functional foods as well as health promoting and pharmaceutical agents.