ABSTRACT
Menopause induces a loss of bone as a result of estrogen deficiency. Despite pharmaceutical options for the treatment of osteopenia and osteoporosis, many aging women use dietary supplements with estrogenic activity to prevent bone loss and other menopausal-related symptoms. Such supplements are yet to be tested for efficacy against a Food and Drug Administration (FDA) approved medication for menopausal bone loss such as zoledronic acid (ZA). The postmenopausal rat model was used to investigate the efficacy of various synergistic phytochemical blends mixed into the diet for 16 weeks. Retired-breeder, Fischer 344 rats were randomly assigned to sham or ovariectomy surgery and 4 treatment groups: ZA; genistein supplementation; and a low dose and high dose blend of genistein, resveratrol, and quercetin. Ovariectomy resulted in a loss of both trabecular and cortical bone which was prevented with ZA. The phytochemical blends tested were unable to reverse these losses. Despite the lack of effectiveness in preventing bone loss, a significant dose-response trend was observed in the phytochemical-rich diets in bone adipocyte number compared to ovariectomized control rats. Data from this study indicate that estrogenic phytochemicals are not as efficacious as ZA in preventing menopausal-related bone loss but may have beneficial effects on bone marrow adiposity in rats.
Subject(s)
Osteoporosis, Postmenopausal/drug therapy , Phytochemicals/administration & dosage , Adiposity/drug effects , Animals , Bone Density/drug effects , Drug Synergism , Drug Therapy, Combination , Female , Genistein/administration & dosage , Humans , Osteoporosis, Postmenopausal/etiology , Osteoporosis, Postmenopausal/physiopathology , Ovariectomy/adverse effects , Quercetin/administration & dosage , Rats , Rats, Inbred F344 , Resveratrol/administration & dosageABSTRACT
OBJECTIVE: Menopausal reduction in estrogen causes increased adipose accumulation, leading many to turn to dietary supplements to prevent and treat such changes. Enhanced adipose mobilization stimulated by some supplements can increase the risk of non-alcoholic fatty liver disease (NAFLD). Cytoprotective and anti-obesity compounds may prevent the lipotoxicity associated with mobilization. METHODS: A phytochemical blend was tested in aged, ovariectomized rats. Rats were given the AIN-93M basal diet or a diet containing varying doses of phytochemicals with 2.4 IU/g vitamin D [diet 1: 1000 mg/kg genistein (G); diet 2: 500 mg/kg (G), 200 mg/kg resveratrol (R), and 1000 mg/kg quercetin (Q); diet 3: 1000 mg/kg (G), 400 mg/kg (R), and 2000 mg/kg (Q)]. RESULTS: Serum free fatty acids and hepatic triglycerides were elevated with diets 2 and 3. Despite this increase, the phytochemical blends did not increase apoptotic, cell repair, or remodeling gene expression. The highest phytochemical dose prevented increases in serum alanine aminotransferase. CONCLUSIONS: Adverse hepatic effects secondary to ovariectomy were mitigated through the inclusion of a dietary phytochemical blend in aged ovariectomized rats. The use of such compounds may not only help with weight management and disease risk in menopausal women, but may also prevent the lipotoxicity in NAFLD.
Subject(s)
Adipose Tissue/drug effects , Adipose Tissue/metabolism , Diet , Fatty Liver/prevention & control , Lipid Metabolism/drug effects , Phytochemicals/pharmacology , Adiposity/drug effects , Animals , Dietary Supplements , Fatty Liver/metabolism , Female , Genistein/pharmacology , Liver/drug effects , Liver/metabolism , Obesity/complications , Obesity/diet therapy , Obesity/metabolism , Ovariectomy , Rats , Rats, Inbred F344 , Triglycerides/metabolismABSTRACT
High-fat diets (HFD) promote the development of both obesity and fatty liver disease through the up-regulation of hepatic lipogenesis. Insulin resistance, a hallmark of both conditions, causes dysfunctional fuel partitioning and increases in lipogenesis. Recent work has demonstrated that systemic insulin resistance occurs in as little as the first 72 hours of an HFD, suggesting the potential for hepatic disruption with HFD at this time point. The current study sought to determine differences in expression of lipogenic genes between sexes in 3-month-old male and female Long-Evans rats after 72 hours of a 40% HFD or a 17% fat (chow) diet. Owing to the response of estrogen on hepatic signaling, we hypothesized that a sexual dimorphic response would occur in the expression of lipogenic enzymes, inflammatory cytokines, apoptotic, and cell repair and remodeling genes. Both sexes consumed more energy when fed an HFD compared with their low fat-fed controls. However, only the males fed the HFD had a significant increase in body fat. Regardless of sex, HFD caused down-regulation of lipogenic and inflammatory genes. Interestingly, females fed an HFD had up-regulated expression of apoptotic and cell repair-related genes compared with the males. This may suggest that females are more responsive to the acute hepatic injury effects caused by HFDs. In summary, neither male nor female rats displayed disrupted hepatic metabolic pathways after 72 hours of the HFD treatment. In addition, female rats appear to have protection from increases in fat deposition, possibly due to increased caloric expenditure; male rats fed an HFD were less active, as demonstrated by distance traveled in their home cage.
Subject(s)
Diet, High-Fat/adverse effects , Gene Expression Regulation , Inflammation/genetics , Lipogenesis/genetics , Liver/metabolism , Animals , Apoptosis/genetics , Body Composition , DNA-Binding Proteins/genetics , Energy Intake , Female , Male , Physical Exertion , Rats , Rats, Long-Evans , Regulatory Factor X Transcription Factors , Sex Factors , Signal Transduction/genetics , Transcription Factors/genetics , Weight Gain , X-Box Binding Protein 1ABSTRACT
Adenovirus 36 (Ad36) is the only adenovirus to date that has been linked with obesity in humans. Our previous studies in late-adolescent females suggest that excess weight in the form of fat mass is associated with lower cortical bone strength. The purpose of this study was to assess the relationship between Ad36-specific antibodies, adiposity, and bone strength in our sample of late-adolescent females. A cross-sectional study of 115 females aged 18 to 19 years was performed. Participants were classified according to adiposity by dual-energy X-ray absorptiometry (body fat percentage as normal-fat [ < 32% body fat; n = 93] or high-fat [ ≥ 32% body fat; n = 22]), and according to the presence of Ad36-specific neutralizing antibodies. Peripheral quantitative computed tomography measured bone parameters at the 4% (trabecular bone) and 20% (cortical bone) site, and muscle cross-sectional area (MCSA) at the 66% site, from the distal metaphyses of the radius and the tibia. Bone strength was determined from volumetric bone mineral density and bone geometry to calculate bone strength index (BSI; trabecular site) and polar strength-strain index (SSI; cortical site). After adjustment for MCSA and limb length, radial SSI was lower in Ad36+ versus Ad36- subjects from the high-fat group (p < 0.03), but not the normal-fat group. No significant differences were observed between groups in tibial SSI or BSI. These data support an association of adiposity and cortical bone strength at the radius with the presence of neutralizing antibodies to Ad36 in late-adolescent females.
Subject(s)
Adenoviridae/pathogenicity , Adiposity , Bone and Bones/physiology , Absorptiometry, Photon , Adolescent , Adult , Female , Humans , Tomography, X-Ray Computed , Young AdultABSTRACT
Estrogens have been shown to protect against various diseases and disastrous metabolic consequences of poor diets. Although a large body of research demonstrates estrogen's ability to control food intake, adipogenesis, and oxidative stress, research regarding the effects of estrogens on hepatic lipogenesis, steatosis, and non-alcoholic fatty liver disease is only now accumulating. Estrogen deficiency in both human and rodent models directly results in the upregulation of hepatic lipogenic signaling - in both serum and hepatic triglyceride content - which leads to the development of fatty liver. In all models, estrogen replacement completely reverses these outcomes. Similar to the endogenous estrogen hormone, plant-derived phytoestrogens also appear to have beneficial effects related to prevention of hepatic lipogenic signaling and steatosis in rodent models. Additionally, such compounds can completely overcome the hepatic consequences that result from estrogen deficiency. While published research strongly supports that estrogens, both endogenous and exogenous, can protect against hepatic lipogenic signaling that can contribute to the development of non-alcoholic fatty liver diseases and adverse weight gain, little research exists on elucidating the mechanism behind this protection. Various pathways have been suggested, including manipulation of both leptin and signal transducer and activator of transcription 3 (STAT3) signaling. However, the discovery of x-box protein 1 elicits the identification of another potential pathway through which estrogen may be working. While the supportive work is strong, further research is needed to determine the mechanism behind the protection by estrogens from hepatic lipogenesis and associated diseases.
ABSTRACT
Vitamin D and certain natural compounds have been shown to regulate both lipid metabolism and bone formation. Treatments that prevent or reverse age-related increase in bone marrow adiposity could both increase new bone formation and inhibit bone destruction. We tested the hypothesis that dietary supplementation with combinations of vitamin D and phytochemicals inhibits bone loss and decreases adiposity to a greater extent than control or vitamin D-alone diets. Aged ovariectomized female rats (12 months old, n=50, initial body weight=240 g) were given control (AIN-93M diet), vitamin D (2,400 IU/kg), or vitamin D plus resveratrol (16, 80, or 400 mg/kg of diet [low, medium, and high dose, respectively]), quercetin (80, 400, or 2,000 mg/kg of diet), and genistein (64, 256, or 1,040 mg/kg of diet) for 8 weeks. The high-dose treatment (vitamin D+400 mg/kg resveratrol+2,000 mg/kg quercetin+1,040 mg/kg genistein) reduced body weight gain (P<.05) and the fat pad weights (P<.05). This treatment also increased the serum concentration of insulin-like growth factor-1 (P<.05) and the bone mineral content of the femur. Micro-computed tomography and histomorphometric analyses indicated that the high-dose treatment prevented loss of trabecular bone (P<.05) and reduced marrow adipocytes (P<.001) and osteoclasts (P<.05) compared with the control and vitamin D alone (P<.05). We conclude that aged ovariectomized female rats supplemented with vitamin D combined with genistein, quercetin, and resveratrol had improved bone mineral density and reduced body weight gain and a significant decrease in bone marrow adipocytes. The synergistic effects of a combination of phytochemicals with vitamin D may be effective in reducing bone loss and weight gain after menopause.
Subject(s)
Bone Diseases, Metabolic/prevention & control , Dietary Supplements , Drug Combinations , Phytotherapy/methods , Vitamin D/administration & dosage , Weight Gain/drug effects , Adipose Tissue/drug effects , Adiposity/drug effects , Animals , Bone Density/drug effects , Diet , Female , Femur/drug effects , Genistein/administration & dosage , Ovariectomy , Quercetin/administration & dosage , Rats , Rats, Inbred F344 , Resveratrol , Stilbenes/administration & dosageABSTRACT
Resveratrol, a phytoalexin, has gained much attention recently due to its effects on sirtuins. While the anti-cancer properties of resveratrol have been extensively investigated, the anti-adipogenic and osteogenic effects of resveratrol are also gaining considerable interest. The finding that resveratrol supplementation mimics caloric restriction prompted researchers to study the effects of resveratrol on lipid metabolism. Mesenchymal stem cells are the precursors for both adipocytes and osteoblasts. In the aging population, differentiation to adipocytes dominates over the differentiation to osteoblasts in bone marrow, contributing to the increased tendency for fractures to occur in the elderly. Thus, an inverse relationship exists between adipocytes and osteoblasts in the bone marrow. Resveratrol acts on several molecular targets in adipocytes and osteoblasts leading to a decrease in adipocyte number and size and an increase in osteogenesis. Furthermore, resveratrol in combination with genistein and quercetin synergistically decreased adipogenesis in murine and human adipocytes. A recent in vivo study showed that phytochemicals including resveratrol in combination with vitamin D prevented weight gain and bone loss in a postmenopausal rat model. Therefore, combinations of resveratrol with other phytochemicals may lead to potential novel potent therapies for both obesity and osteoporosis.
Subject(s)
Obesity/drug therapy , Osteoporosis/drug therapy , Stilbenes/therapeutic use , Adipocytes/drug effects , Adipocytes/physiology , Animals , Caloric Restriction , Drug Synergism , Humans , Lipolysis/drug effects , Osteoblasts/physiology , Resveratrol , Stilbenes/pharmacologyABSTRACT
Both central and peripheral leptin administrations reduce body weight, food intake, and adiposity in ob/ob mice. In this study we compared effects of intracerebroventricular (ICV) and subcutaneous (SC) administration of leptin on bone metabolism in the appendicular and axial skeleton and adipose tissue gene expression and determined the effects of ICV leptin on bone marrow gene expression in ob/ob mice. In experiment 1, leptin (1.5 or 0.38 µg/d) or control was continuously injected ICV for 12 days. Gene expression analysis of femoral bone marrow stromal cells showed that expression of genes associated with osteogenesis was increased after ICV injection, whereas those associated with osteoclastogenesis, adipogenesis, and adipocyte lipid storage were decreased. In experiment 2, leptin was injected continuously ICV (0.0 or 1.5 µg/d) or SC (0.0 or 10 µg/d) for 12 days. In both experiments, regardless of mode of administration, leptin decreased body weight, food intake, and body fat and increased muscle mass, bone mineral density, bone mineral content, bone area, marrow adipocyte number, and mineral apposition rate. Serum insulin was decreased, whereas serum osteocalcin, insulin-like growth factor 1, osteoprotegerin, pyridinoline, and receptor activator of nuclear factor κB ligand concentrations were increased. In experiment 2, expression of genes in adipose tissue associated with apoptosis, lipid mobilization, insulin sensitivity, and thermogenesis was increased, whereas expression of genes associated with cell differentiation and maturation was decreased regardless of mode of administration. Thus ICV injection of leptin promotes expression of pro-osteogenic factors in bone marrow, leading to enhanced bone formation in ob/ob mice.
Subject(s)
Bone Density/drug effects , Insulin-Like Growth Factor I/metabolism , Leptin/deficiency , Leptin/pharmacology , Muscles/anatomy & histology , Osteogenesis/drug effects , Osteogenesis/genetics , Adipocytes/cytology , Adipocytes/drug effects , Adipocytes/metabolism , Animals , Body Weight/drug effects , Bone Marrow/drug effects , Bone Marrow/metabolism , Cell Count , Feeding Behavior/drug effects , Female , Femur/drug effects , Femur/metabolism , Gene Expression Regulation/drug effects , Injections, Intraventricular , Leptin/administration & dosage , Mice , Mice, Inbred C57BL , Mice, Obese , Muscles/drug effects , Organ Size/drug effects , Tibia/anatomy & histology , Tibia/drug effects , Tibia/metabolismABSTRACT
Evidence from both epidemiological studies and basic research suggests that obesity and osteoporosis are interrelated. Though there is an increase in the prevalence of these disorders, a limited number of treatments are available, one of the reasons being the complexity of the pathways involved and difficulty in identifying a single molecular target. Due to adverse effects of pharmaceuticals, intake of herbal drugs by patients without a physician's recommendation is increasing globally. Lack of success with targeted monotherapy has encouraged scientists to determine whether combinations of phytochemicals that interfere with numerous cell-signaling pathways can be a more effective approach to treat complex diseases. For example, evidence is emerging that specific combinations of phytochemicals are far more effective than single compounds in decreasing adipogenesis and promoting bone formation. Since multiple pathways are dysfunctional in obesity and osteoporosis, an ideal approach for preventing and treating these diseases may be to use a combination of phytochemicals to address several targets simultaneously.
Subject(s)
Obesity/prevention & control , Osteoporosis/prevention & control , Plant Extracts/pharmacology , Adipocytes/drug effects , Adipocytes/metabolism , Adipogenesis/physiology , Humans , Lipogenesis/physiology , Models, Biological , Obesity/drug therapy , Osteoblasts/metabolism , Osteogenesis/physiology , Osteoporosis/drug therapy , Phytotherapy/methods , Signal TransductionABSTRACT
Higher levels of body fat are associated with increased risk for development of numerous adverse health conditions. Phytochemicals are potential agents to inhibit differentiation of preadipocytes, stimulate lipolysis, and induce apoptosis of existing adipocytes, thereby reducing adipose tissue mass. Resveratrol decreased adipogenesis and viability in maturing preadipocytes; these effects were mediated not only through down-regulating adipocyte specific transcription factors and enzymes but also by genes that modulate mitochondrial function. Additionally, resveratrol increased lipolysis and reduced lipogenesis in mature adipocytes. In addition, combining resveratrol with other natural products produced synergistic activities from actions on multiple molecular targets in the adipocyte life cycle. Treatment of mice with resveratrol alone was shown to improve resistance to weight gain caused by a high-fat diet. Moreover, dietary supplementation of aged ovariectomized rats with a combination of resveratrol and vitamin D, quercetin, and genistein not only decreased weight gain but also inhibited bone loss. Combining several phytochemicals, including resveratrol, or using them as templates for synthesizing new drugs, provides a large potential for using phytochemicals to target adipocyte adipogenesis, apoptosis, and lipolysis.
Subject(s)
Adipogenesis/drug effects , Adipose Tissue/drug effects , Stilbenes/pharmacology , Adipogenesis/physiology , Adipose Tissue/metabolism , Animals , Humans , Lipolysis/drug effects , Lipolysis/physiology , Obesity/drug therapy , Obesity/metabolism , Resveratrol , Signal Transduction/drug effects , Signal Transduction/physiology , Stilbenes/therapeutic useABSTRACT
Comparative proteomic analyses were performed in adipose tissue of leptin-deficient ob/ob mice treated with leptin or control buffer in order to identify the protein expression changes as the potential targets of leptin. Mice were treated with either phosphate-buffered saline (control) or 10 µg/day leptin for 14 days via subcutaneous osmotic minipumps. Total protein from white adipose tissue was extracted and labeled with different fluorescent cyanine dyes for analysis by two-dimensional difference gel electrophoresis (DIGE). Spots that were differentially expressed and appeared to have sufficient material for mass spectrometry analysis were picked and digested with trypsin and subjected to MALDI-TOF MS for protein identification. Twelve functional protein groups were found differentially expressed in adipose tissue of leptin-treated vs. control ob/ob mice, including molecular chaperones and redox proteins such as calreticulin (CALR), protein disulfide isomerase-associated 3 (PDIA3), prohibitin (PHB), and peroxiredoxin-6 (PRDX6); cytoskeleton proteins such as ß actin, desmin, and α-tubulin; and some other proteins. The mRNA levels of CALR, PDIA3, and PHB were measured by real-time reverse transcription-PCR and found to be upregulated (P < 0.05), consistent with the fold change in protein expression level. Our findings suggest that leptin's effects on lipid metabolism and apoptosis may be mediated in part by alterations in expression of molecular chaperones and redox proteins for regulating endoplasmic reticulum stress and cytoskeleton proteins for regulating mitochondrial morphology.
Subject(s)
Adipose Tissue/metabolism , Apoptosis Regulatory Proteins/metabolism , Leptin/deficiency , Leptin/pharmacology , Lipid Metabolism/drug effects , Animals , Apoptosis Regulatory Proteins/drug effects , Apoptosis Regulatory Proteins/genetics , Gene Expression Profiling , Humans , Leptin/genetics , Leptin/metabolism , Mice , Mice, Obese , Obesity/genetics , Obesity/metabolism , Prohibitins , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Two-Dimensional Difference Gel ElectrophoresisABSTRACT
No disponible
Clenbuterol, a beta2-adrenergic receptor (Beta2-AR) selective agonist, has been shown to decrease body fat in animals and can induce apoptosis in adipose tissue in mice. We hypothesized that direct actions of a Beta -adrenergic receptor agonist on adipocytes could trigger the observed apoptotic effect. The hypothesis was inspected by investigating the direct effect of clenbuterol on apoptosis,adipogenesis, and lipolysis in vitro using the 3T3-L1 cell line and rat primary adipocytes. Cells were treated with ( ) (AU)
Subject(s)
Animals , Mice , Clenbuterol/pharmacokinetics , Apoptosis , Adipogenesis , Lipolysis , Adipocytes , Adrenergic beta-Agonists/pharmacokinetics , Adiposity , 3T3 CellsABSTRACT
Obesity is an increasing health problem all over the world. Phytochemicals are potential agents to inhibit differentiation of preadipocytes, stimulate lipolysis, and induce apoptosis of existing adipocytes, thereby reducing the amount of adipose tissue. Flavonoids and stilbenoids represent the most researched groups of phytochemicals with regards to their effect on adipogenesis, but there are also a number of in vitro and in vivo studies with phenolic acids, alkaloids, and vitamins, as well as other plant compounds. Although phytochemicals like epigallocatechin-3-gallate, genistein, and resveratrol reduce lipid accumulation and induce adipocyte apoptosis in vitro and reduce body weight and adipose tissues mass in animal models of diet-induced obesity, well-conducted clinical trials are lacking. Pharmacological doses are often used in vitro and when applied in physiological doses in animals or humans, the phytochemicals are often ineffective in affecting adipogenesis. However, by combining several phytochemicals or using them as templates for synthesizing new drugs, there is a large potential in targeting adipogenesis using phytochemicals.
Subject(s)
Adipocytes/metabolism , Adipogenesis/drug effects , Apoptosis/drug effects , Lipid Metabolism/drug effects , Lipolysis/drug effects , Obesity/drug therapy , Phytotherapy , Plant Extracts/pharmacology , Plant Extracts/therapeutic use , Animals , Cell Proliferation/drug effects , Humans , Mice , RatsABSTRACT
Clenbuterol, a beta(2)-adrenergic receptor (beta(2)-AR) selective agonist, has been shown to decrease body fat in animals and can induce apoptosis in adipose tissue in mice. We hypothesized that direct actions of a beta-adrenergic receptor agonist on adipocytes could trigger the observed apoptotic effect. The hypothesis was inspected by investigating the direct effect of clenbuterol on apoptosis, adipogenesis, and lipolysis in vitro using the 3T3-L1 cell line and rat primary adipocytes. Cells were treated with 10(-9) to 10(-5) M clenbuterol depending on the experiments. There was no apoptotic effect of clenbuterol both in 3T3-L1 cells and rat primary adipocytes. Adipogenesis monitored by Oil Red O staining and AdipoRed assay was modestly decreased by clenbuterol treatment (p < 0.05). In fully differentiated primary adipocytes, clenbuterol increased basal lipolysis compared with the control (p < 0.01). In summary, direct stimulation of beta(2)-AR by clenbuterol does not cause apoptosis in adipocytes, despite a direct lipolytic stimulation and attenuation of adipogenesis.
Subject(s)
Adipocytes/drug effects , Adipogenesis/drug effects , Adrenergic beta-Agonists/pharmacology , Apoptosis/drug effects , Clenbuterol/pharmacology , Lipolysis/drug effects , Adipocytes/metabolism , Animals , Cell Survival/drug effects , Mice , RatsABSTRACT
Excessive gestational weight gain and maternal obesity have both been associated with increased incidence of obesity and metabolic disorder in offspring in both humans and animal models. The objectives of this study were to determine (1) whether mild gestational food restriction during the third trimester (GFR) would alter food intake and growth parameters of offspring, (2) whether effects of GFR depended on diet (high fat [HF] vs chow), (3) whether effects of excessive gestational weight gain (WG) would become magnified across generations, and (4) whether diet and GFR would alter hypothalamic gene expression in adult offspring. Three generations of female C57BL/6 mice were fed chow or HF diet, mated at 11 weeks of age and assigned to ad libitum feeding or 25% GFR. Offspring were fed the same diet as their mothers. Results showed (1) maternal gestational WG was positively correlated with offspring WG. (2) HF offspring weighed less (p<0.01) at weaning (WWT) but gained more during the 8 weeks after weaning than chow-fed offspring (p<0.05), resulting in higher final body weights (BW) (p<0.01). (3) HF males from GFR mothers had higher WWT (p<0.05), but subsequent WG and final BW were less (p<0.05) compared to males from ad lib mothers. (4) In the HF group, GFR also resulted in decreased FI (p<0.05) and FE (p<0.07) in offspring, compared to offspring from ad lib mothers. (5) In generation 3, hypothalamic expression of tyrosine hydroxylase was lower in HF males from GFR mothers compared to HF males from ad lib mothers (p<0.05). In conclusion, gender and maternal GFR had independent effects on growth and FI, and hypothalamic gene expression was dependent on both gender and maternal GFR in HF offspring. Even mild food restriction of obese mothers during pregnancy may have beneficial effects in reducing the risk or degree of obesity in offspring.
Subject(s)
Body Weight/physiology , Dietary Fats/administration & dosage , Eating/physiology , Gene Expression Regulation, Developmental/physiology , Hypothalamus/physiology , Maternal Nutritional Physiological Phenomena/physiology , Prenatal Exposure Delayed Effects/metabolism , Age Factors , Animals , Birth Weight/physiology , Caloric Restriction/methods , Female , Male , Mice , Mice, Inbred C57BL , Obesity/etiology , Obesity/metabolism , Pregnancy , Sex Factors , Weight Gain/physiologyABSTRACT
This study compared the central effects of ghrelin and leptin on body and bone marrow adiposity and gene expression in adipose tissue and bone marrow. Male Sprague-Dawley rats were injected intracerebroventricular (ICV) twice daily with control, 66 ng ghrelin (G66), 330 ng ghrelin (G330), or 5 µg leptin (L5) for 5 days. Food intake (FI) and body weight (BW) were measured daily. Gene expression in adipose tissue and bone marrow was assessed using RT-PCR. Leptin reduced FI (P < 0.05) and BW (P < 0.05), whereas ghrelin increased BW (P < 0.05) without affecting FI. Leptin decreased fat pad weights, whereas ghrelin (G330) increased fat pad weights (P < 0.05). In epididymal adipose tissue, leptin increased expression of lipolysis marker ADRB2 and thermogenesis marker MFN2 and decreased expression of adipogenic markers, FASN, SLC2A4, and SCD1, whereas ghrelin increased expression of FASN and SCD1. Leptin decreased bone marrow adipocyte size and number; however, ghrelin had no effect on these parameters. In whole bone marrow, leptin decreased expression of FASN and SCD1 and increased expression of DLK1, whereas ghrelin (G330) decreased expression of COL1A1. Thus, leptin induces similar changes in bone marrow and adipose tissue gene expression, reflecting the decreased adiposity in both compartments.
Subject(s)
Adipose Tissue, White/metabolism , Adiposity , Bone Marrow/metabolism , Gene Expression Regulation , Ghrelin/physiology , Leptin/physiology , Adipogenesis , Adipose Tissue, White/cytology , Animals , Biomarkers , Body Weight , Bone Marrow Cells/cytology , Bone Marrow Cells/metabolism , Cells, Cultured , Diet , Gene Expression Profiling , Ghrelin/administration & dosage , Injections, Intraventricular , Leptin/administration & dosage , Lipolysis , Male , RNA, Messenger/metabolism , Random Allocation , Rats , Rats, Sprague-Dawley , ThermogenesisABSTRACT
The effect of octanoate and decanoate, respectively, eight- and 10-carbon medium-chain fatty acids (MCFAs), on apoptotic signaling in 3T3-L1 adipocytes was investigated. 3T3-L1 adipocytes were treated with various concentrations of octanoate or decanoate. Cell viability, apoptosis, and expression of apoptosis-related proteins were investigated. Results indicated that both octanoate and decanoate decreased viability, increased apoptosis, and increased reactive oxygen species production. Immunoblotting analysis showed an increase in the levels of cytoplasmic cytochrome c and cleaved poly(ADP-ribose) polymerase by octanoate and decanoate. Concomitantly, we observed that pro-caspase-3 was decreased, resulting in the induced accumulation of the cleaved form of caspase-3 by both octanoate and decanoate. In addition, both octanoate and decanoate increased the expression of pro-apoptotic Bax with an accompanied decrease of anti-apoptotic Bcl-2. These results show that octanoate and decanoate mediate adipocyte apoptosis via a caspase-dependent mitochondrial pathway in 3T3-L1 adipocytes. MCFAs thus decrease adipocyte number by initiating the apoptotic process in 3T3-L1 adipocytes.
Subject(s)
Adipocytes/drug effects , Apoptosis/drug effects , Caprylates/pharmacology , Caspase 3/metabolism , Cell Survival/drug effects , Decanoates/pharmacology , Reactive Oxygen Species/metabolism , 3T3-L1 Cells , Adipocytes/physiology , Animals , Cytochromes c/metabolism , Mice , Mitochondria/drug effects , Poly(ADP-ribose) Polymerases/metabolism , Proto-Oncogene Proteins c-bcl-2/metabolism , Signal Transduction/drug effects , bcl-2-Associated X Protein/metabolismABSTRACT
Xanthohumol (XN) and guggulsterone (GS) have each been shown to inhibit adipogenesis and induce apoptosis in adipocytes. In the present study effects of the combination of XN + GS on 3T3-L1 adipocyte apoptosis and adipogenesis were investigated. Mature adipocytes were treated with XN and GS individually and in combination. XN and GS individually decreased cell viability, but XN + GS caused an enhanced decrease in viability and potentiated induction of apoptosis. Likewise, XN + GS caused a potentiated increase in caspase-3/7 activation, whereas neither of the compounds showed any effect individually. In addition, western blot analysis revealed that XN + GS increased Bax expression and decreased Bcl-2 expression, whereas individual compounds did not show any significant effect. XN and GS both decreased lipid accumulation. Individually, XN at 1.5 microM and GS at 3.12 microM decreased lipid accumulation by 26 +/- 4.5% (P < .001) each, whereas XN1.5 + GS3.12 decreased lipid accumulation by 78.2 +/- 1.8% (P < .001). Moreover, expression of the adipocyte-specific proteins was down-regulated with XN1.5 + GS3.12, but no effect was observed with the individual compounds. Finally, XN + GS caused an enhanced stimulation of lipolysis. Thus, combination of XN and GS is more potent in exerting anti-obesity effects than additive effects of the individual compounds.
Subject(s)
Adipocytes/drug effects , Adipogenesis/drug effects , Anti-Obesity Agents/pharmacology , Apoptosis/drug effects , Flavonoids/pharmacology , Lipid Metabolism/drug effects , Pregnenediones/pharmacology , Propiophenones/pharmacology , 3T3-L1 Cells , Adipocytes/metabolism , Animals , Anti-Obesity Agents/therapeutic use , Caspases/metabolism , Cell Survival/drug effects , Commiphora/chemistry , Down-Regulation , Drug Therapy, Combination , Flavonoids/therapeutic use , Gene Expression , Lipolysis/drug effects , Mice , Obesity/metabolism , Obesity/prevention & control , Phytotherapy , Plant Extracts/pharmacology , Plant Extracts/therapeutic use , Propiophenones/therapeutic use , Proto-Oncogene Proteins c-bcl-2/genetics , Proto-Oncogene Proteins c-bcl-2/metabolism , bcl-2-Associated X Protein/genetics , bcl-2-Associated X Protein/metabolismABSTRACT
We report here that octanoate and decanoate, 8-carbon and 10-carbon medium-chain fatty acids (MCFA), decreased adipogenesis in 3T3-L1 preadipocytes when treated with standard hormonal cocktail, but increased adipogenesis in a dose-dependent manner (with decanoate being more effective) when treated with basal media. Addition of dexamethasone to basal medium with either octanoate or decanoate further increased adipogenesis. In order to understand the adipogenic effects of MCFA in the absence of standard hormonal cocktail, postconfluent 3T3-L1 preadipocytes were treated with octanoate or decanoate, and the change in the expression of several adipogenic transcription factors and enzymes was investigated using real-time RT-PCR. Octanoate and decanoate up-regulated the mRNA expression of peroxisome-proliferator-activated receptor (PPAR) gamma, CCAAT/enhancer-binding protein (C/EBP) alpha, fatty-acid-binding protein, sterol-regulatory element binding protein 1c, lipoprotein lipase and hormone-sensitive lipase, and the protein expression of PPARgamma and C/EBPalpha, with decanoate being more effective. Moreover, the PPARgamma antagonist GW9662 inhibited MCFA-induced lipid accumulation by about 50%. Decanoate and octanoate, to a lesser degree, increased lipid accumulation, which was associated with an increase in glycerol-3-phosphate dehydrogenase activity. These results show that octanoate and decanoate may stimulate differentiation of preadipocytes, at least in part, by their influence on the expression of PPARgamma and other adipocyte-specific factors.
Subject(s)
Adipogenesis/drug effects , Caprylates/pharmacology , Decanoates/pharmacology , 3T3-L1 Cells , Adipocytes/cytology , Adipocytes/drug effects , Adipocytes/metabolism , Anilides/pharmacology , Animals , Anisoles/pharmacology , CCAAT-Enhancer-Binding Protein-alpha/metabolism , Dexamethasone/pharmacology , Gene Expression/drug effects , Glycerolphosphate Dehydrogenase/genetics , Glycerolphosphate Dehydrogenase/metabolism , Hormones/pharmacology , Lipid Metabolism , Mice , PPAR gamma/antagonists & inhibitors , Transcription Factors/metabolismABSTRACT
This paper describes effects of several sulfur-containing compounds from garlic on the cell viability, apoptosis and adipogenesis in 3T3-L1 adipocytes. In both preadipocytes and mature adipocytes, 100 and 200 microM ajoene significantly decreased cell viability and increased apoptosis. The effect on apoptosis was further confirmed with Hoechst staining. In contrast, diallyl sulfide, diallyl disulfide, diallyl trisulfide, deoxyalliin, and allyl methyl sulfide had no significant effect on cell viability or apoptosis in either preadipocytes or mature adipocytes. In maturing preadipocytes ajoene significantly decreased lipid accumulation in a dose-dependent manner and these results were further confirmed by a decrease in lipid droplet number and lipid content through Oil Red O staining. There was no significant change in lipid accumulation in maturing preadipocytes treated with other garlic derivatives. Thus, despite the same source of origin, garlic, ajoene was the only one with potent effects on cell viability, apoptosis and adipogenesis in 3T3-L1 adipocytes.