ABSTRACT
The present study was performed to investigate the molecular mechanism of 6-gingerol on adipocyte-mediated systemic inflammation in vitro and in high-fat diet-induced obese zebra fish. 6-Gingerol decreased adipogenesis due to the suppression of adipocyte differentiation markers, including peroxisome proliferator-activated receptor gamma, CCAATT enhancer binding protein α, and adipocyte protein 2, and triglyceride synthesis enzymes, including sterol regulatory element-binding protein-1, fatty acid synthase, lysophosphatidic acid acyltransferase, and acyl-coAâ:âdiacylglycerol acyltransferase 1, in 3T3-L1. A coculture insert system using 3T3-L1 with RAW 264.7 (coculture insert system using fully differentiated 3T3-L1 cells with RAW 264.7 macrophages) revealed that 6-gingerol increased anti-inflammatory cytokine interleukin-10. The expression of TNFα, monocyte chemotactic protein-1, interleukin-1ß, and interleukin-6 were decreased in the coculture insert system using fully differentiated 3T3-L1 cells with RAW 264.7 macrophages treated with 6-gingerol. Moreover, the coculture insert system using fully differentiated 3T3-L1 cells with RAW 264.7 macrophages treated with 6-gingerol inhibited the protein expression of TNFα and monocyte chemotactic protein-1 in RAW 264.7. 6-Gingerol decreased c-JUN N-terminal kinase and I kappa B kinase beta and its downstream target AP-1 expression in the coculture insert system using fully differentiated 3T3-L1 cells with RAW 264.7 macrophages. Furthermore, 6-gingerol decreased the expression of inducible nitric oxide synthase stimulated by the coculture insert system using fully differentiated 3T3-L1 cells with RAW 264.7 macrophages in RAW 264.7 and attenuated nitric oxide production in diet-induced obese zebra fish. Our results suggest that 6-gingerol suppresses inflammation through the regulation of the c-JUN N-terminal kinase-I kappa B kinase beta and its downstream targets.
Subject(s)
Adipocytes/drug effects , Catechols/pharmacology , Fatty Alcohols/pharmacology , Inflammation/drug therapy , Inflammation/metabolism , Obesity/drug therapy , Obesity/metabolism , 3T3-L1 Cells , Acyltransferases/metabolism , Adipocytes/cytology , Adipocytes/metabolism , Adipogenesis/drug effects , Animals , Cytokines/metabolism , Diacylglycerol O-Acyltransferase/metabolism , Diet, High-Fat , Down-Regulation/drug effects , Fatty Acid Synthases/metabolism , Fatty Acid-Binding Proteins/metabolism , I-kappa B Kinase/metabolism , In Vitro Techniques , Inflammation/pathology , JNK Mitogen-Activated Protein Kinases/metabolism , Macrophages/drug effects , Macrophages/metabolism , Mice , Nitric Oxide/metabolism , Obesity/pathology , PPAR gamma/drug effects , RAW 264.7 Cells , Sterol Regulatory Element Binding Protein 1/metabolism , Transcription Factor AP-1/metabolism , Triglycerides/metabolism , ZebrafishABSTRACT
Fucoxanthin, a pigment from the chloroplasts of marine brown algae, has a number of effects against obesity, diabetes, inflammation and cancer and provides cerebrovascular protection. In this study, we investigated the inhibitory effects of fucoxanthin on lipid accumulation and reactive oxygen species (ROS) production during adipogenesis. Treatment with fucoxanthin suppresses protein levels of the adipogenic transcription factors CCAAT/enhancer-binding protein alpha C/EBPα and peroxisome proliferator-activated receptor-γ and of their target protein, fatty acid binding protein 4. Lipogenesis-related enzymes, such as diglyceride acyltransferase 1 and lysophosphatidic acid acyltransferase-θ, were downregulated by fucoxanthin. The ROS-producing enzyme nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 4 (NOX4) and the NADPH-generating enzyme glucose-6-phosphate dehydrogenase also decreased following fucoxanthin treatment. The adipokine adiponectin and the ROS-scavenging enzymes superoxide dismutase 2, glutathione reductase and catalase were dose-dependently increased by fucoxanthin. Furthermore, lipolysis-related enzymes and superoxide dismutase 1 were slightly decreased, because of the suppression of lipid-generating factors and the cytosolic enzyme NOX4. To confirm these results, we investigated lipid accumulation and ROS production in zebrafish, where fucoxanthin suppressed lipid and triglyceride accumulation, as well as ROS production. Our data suggest that fucoxanthin inhibits lipid accumulation and ROS production by controlling adipogenic and lipogenic factors and ROS-regulating enzymes. Copyright © 2016 John Wiley & Sons, Ltd.
Subject(s)
3T3-L1 Cells/metabolism , Adipocytes/drug effects , Adipogenesis/drug effects , Lipid Metabolism/drug effects , Xanthophylls/chemistry , Animals , Cell Differentiation , Mice , Reactive Oxygen Species , Xanthophylls/pharmacology , ZebrafishABSTRACT
Ellagic acid (EA) is a natural polyphenol found in various fruits and vegetables. In this study, we examined the inhibitory effect of EA on fat accumulation in 3T3-L1 cells during adipogenesis. Our data showed that EA reduced fat accumulation by down-regulating adipogenic markers such as peroxisome proliferator activated receptor γ (PPARγ) and the CCAAT/enhancer binding protein α (C/EBPα) at the mRNA and protein levels in a dose-dependent manner. We found that the decrease in adipogenic markers resulted from reduced expression of some early adipogenic transcription factors such as KLF4, KLF5, Krox20, and C/EBPß within 24 h. Also, these inhibitions were correlated with down-regulation of TG synthetic enzymes, causing inhibition of triglyceride (TG) levels in 3T3-L1 cells investigated by ORO staining and in zebrafish investigated by TG assay. Additionally, the cell cycle analysis showed that EA inhibited cell cycle progression by arresting cells at the G0/G1 phase.
Subject(s)
Adipocytes/drug effects , Adipogenesis/drug effects , Cell Cycle Checkpoints , Ellagic Acid/pharmacology , Lipid Metabolism/drug effects , 3T3-L1 Cells , Animals , CCAAT-Enhancer-Binding Protein-alpha/metabolism , CCAAT-Enhancer-Binding Protein-beta/genetics , Down-Regulation , Kruppel-Like Factor 4 , Mice , PPAR gamma/metabolism , Polyphenols/pharmacology , Transcription Factors/metabolism , ZebrafishABSTRACT
Gelidium elegans is an edible red alga native to the intertidal area of northeastern Asia. We investigated the effect of G. elegans extract and its main flavonoids, rutin and hesperidin, on lipid accumulation and the production of reactive oxygen species (ROS) and reactive nitrogen species (RNS) in 3T3-L1 and RAW264.7 cells. Our data show that G. elegans extract decreased lipid accumulation and ROS/RNS production in a dose-dependent manner. The extract also inhibited the mRNA expression of adipogenic transcription factors, such as peroxisome proliferator-activated receptor gamma and CCAAT/enhancer-binding protein alpha, while enhancing the protein expression of the antioxidant enzymes superoxide dismutases 1 and 2, glutathione peroxidase, and glutathione reductase compared with controls. In addition, lipopolysaccharide-induced nitric oxide production was significantly reduced in G. elegans extract-treated RAW264.7 cells. In analysis of the effects of G. elegans flavonoids on lipid accumulation and ROS/RNS production, only hesperidin showed an inhibitory effect on lipid accumulation and ROS production; rutin did not affect adipogenesis and ROS status. The antiadipogenic effect of hesperidin was evidenced by the downregulation of peroxisome proliferator-activated receptor gamma, CCAAT/enhancer-binding protein alpha, and fatty acid binding protein 4 gene expression. Collectively, our data suggest that G. elegans is a potential food source containing antiobesity and antioxidant constituents.
Subject(s)
Adipocytes/drug effects , Antioxidants/pharmacology , Hesperidin/pharmacology , Reactive Nitrogen Species/metabolism , Reactive Oxygen Species/metabolism , Seaweed/chemistry , 3T3-L1 Cells , Adipogenesis/drug effects , Animals , CCAAT-Enhancer-Binding Protein-alpha/metabolism , Cell Line , Glutathione Peroxidase/metabolism , Glutathione Reductase/metabolism , Lipid Metabolism , Macrophages/drug effects , Mice , Nitric Oxide/metabolism , PPAR gamma/metabolism , Plant Extracts/chemistry , Plant Extracts/pharmacology , Rhodophyta/chemistry , Rutin/pharmacology , Superoxide Dismutase/metabolismABSTRACT
New therapeutic strategies are needed to combat the tuberculosis pandemic and the spread of multidrug-resistant (MDR) and extensively drug-resistant (XDR) forms of the disease, which remain a serious public health challenge worldwide. The most urgent clinical need is to discover potent agents capable of reducing the duration of MDR and XDR tuberculosis therapy with a success rate comparable to that of current therapies for drug-susceptible tuberculosis. The last decade has seen the discovery of new agent classes for the management of tuberculosis, several of which are currently in clinical trials. However, given the high attrition rate of drug candidates during clinical development and the emergence of drug resistance, the discovery of additional clinical candidates is clearly needed. Here, we report on a promising class of imidazopyridine amide (IPA) compounds that block Mycobacterium tuberculosis growth by targeting the respiratory cytochrome bc1 complex. The optimized IPA compound Q203 inhibited the growth of MDR and XDR M. tuberculosis clinical isolates in culture broth medium in the low nanomolar range and was efficacious in a mouse model of tuberculosis at a dose less than 1 mg per kg body weight, which highlights the potency of this compound. In addition, Q203 displays pharmacokinetic and safety profiles compatible with once-daily dosing. Together, our data indicate that Q203 is a promising new clinical candidate for the treatment of tuberculosis.
Subject(s)
Adenosine Triphosphate/biosynthesis , Electron Transport Complex III/antagonists & inhibitors , Extensively Drug-Resistant Tuberculosis/drug therapy , Imidazoles/pharmacology , Mycobacterium tuberculosis/drug effects , Piperidines/pharmacology , Pyridines/pharmacology , Animals , Cell Proliferation/drug effects , Disease Models, Animal , Drug Evaluation, Preclinical , Electron Transport Complex III/genetics , Imidazoles/pharmacokinetics , Mice , Mice, Inbred BALB C , Piperidines/pharmacokinetics , Pyridines/pharmacokinetics , Rats , Rats, Sprague-DawleyABSTRACT
Grateloupia lanceolata (Okamura) Kawaguchi is a red alga native to coastal areas of East Asia. The effect of a G. lanceolata extract on lipid accumulation and reactive oxygen species (ROS) production in 3T3-L1 cells was assessed by examining adipogenic transcription factors and ROS-regulating genes at the molecular level. An ethanol extract of G. lanceolata inhibited lipid accumulation and ROS production during adipogenesis. Treatment with the G. lanceolata extract lead to a reduction in the mRNA levels of the transcription factors, peroxisome proliferator-activated receptor-γ and CCAAT/ enhancer binding protein-α, and at the protein level for the target protein, adipocyte protein 2. ROS-producing nicotinamide adenine dinucleotide phosphate hydrogen (NADPH) oxidase 4 and NADPH-producing glucose-6-phosphate dehydrogenase mRNAs decreased following G. lanceolata extract treatment. In contrast, the mRNA level of ROS scavenging enzymes, including superoxide dismutase (SOD), glutathione peroxidase, and catalase increased in the extract-treated group. The increase in SOD1 (Cu/Zn-SOD) and 2 (Mn-SOD) proteins was correlated with their mRNA levels. Additionally, the G. lanceolata extract significantly enhanced mRNA levels of adiponectin, one of the adipokines secreted from adipocytes. Our results show that G. lanceolata extract inhibited lipid accumulation and ROS production by controlling adipogenic signals and ROS regulating genes.
Subject(s)
Adipogenesis/drug effects , Lipid Metabolism/drug effects , Plant Extracts/pharmacology , Reactive Oxygen Species/metabolism , Rhodophyta/chemistry , Seaweed/chemistry , 3T3-L1 Cells , Animals , Gene Expression Regulation/drug effects , MiceABSTRACT
Pycnogenol® is a group of flavonoids with antioxidant effects. Adipogenesis is the process of adipocyte differentiation. It causes the increase of lipids as well as ROS (reactive oxygen species). Lipid accumulation and ROS production were determined in 3 T3-L1 adipocyte, and the effect of Pycnogenol® was evaluated. Lipid accumulation was elevated in adipocyte treated with hydrogen peroxide, one of the ROS. Pycnogenol® showed an inhibitory effect on the lipid accumulation and ROS production during the adipogenesis. We also investigated the molecular events associated with ROS production and lipid accumulation. Our results showed that Pycnogenol® inhibited the mRNA expression of pro-oxidant enzymes, such as NOX4 (NADPH (nicotinamide adenine dinucleotide phosphate hydrogen) oxidase 4), and the NADPH-producing G6PDH (glucose-6-phosphate dehydrogenase) enzyme. In addition, Pycnogenol® suppressed the mRNA abundance of adipogenic transcription factors, PPAR-γ (peroxisome proliferator-activated receptor γ) and C/EBP-α (CCAAT/enhancer binding protein α), and their target gene, aP2 (adipocyte protein 2) responsible for fatty acid transportation. On the other hand, Pycnogenol® increased the abundance of antioxidant proteins such as Cu/Zn-SOD (copper-zinc superoxide dismutase), Mn-SOD (manganese superoxide dismutase), GPx (glutathione peroxidase) and GR (glutathione reductase). Our results suggest that Pycnogenol® inhibits lipid accumulation and ROS production by regulating adipogenic gene expression and pro-/antioxidant enzyme responses in adipocytes.
Subject(s)
Adipocytes/drug effects , Antioxidants/metabolism , Flavonoids/pharmacology , Lipid Metabolism , Reactive Oxygen Species/metabolism , 3T3-L1 Cells , Adipocytes/cytology , Adipocytes/metabolism , Adipogenesis , Animals , Gene Expression Regulation, Enzymologic , Glucosephosphate Dehydrogenase/genetics , Glucosephosphate Dehydrogenase/metabolism , Glutathione Peroxidase/metabolism , Glutathione Reductase/metabolism , Hydrogen Peroxide/pharmacology , Mice , NADPH Oxidase 4 , NADPH Oxidases/genetics , NADPH Oxidases/metabolism , PPAR gamma/genetics , PPAR gamma/metabolism , Plant Extracts , RNA, Messenger/genetics , RNA, Messenger/metabolism , Superoxide Dismutase/metabolism , Transcription Factors/genetics , Transcription Factors/metabolismABSTRACT
Tocochromanols are potent lipid-soluble antioxidants and essential nutrients for human health. Genetic engineering techniques were used to develop soybeans with enhanced vitamin E levels, including tocotrienols, which are not found in soybean. The gene encoding rice homogentisate geranylgeranyl transferase (HGGT) was overexpressed in soybeans using seed-specific and constitutive promoters. The association between abundance of vitamin E isomers and antioxidant activity was investigated during seed germination. With the exception of ß-tocotrienol, all vitamin E isomers were detected in germinating seeds expressing OsHGGT. The antioxidant properties of germinating seed extracts were determined using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radicals and lipid peroxidation (TBARS). Compared with intact wild-type seeds, transgenic seeds showed increases in radical scavenging of 5.4-17 and 23.2-35.3% in the DPPH and ABTS assays, respectively. Furthermore, the lipid peroxidation levels were 2.0-4.5-fold lower in germinating seeds from transgenic lines than in wild-type seeds. Therefore, it appears that the antioxidant potential of transgenic oil-producing plants such as soybean, sunflower, and corn may be enhanced by overexpressing OsHGGT during seed germination.