Phytochemicals Added to the Feed of Ovariectomized Adult Rats Increase Brown Adipose Activity

Background: Brown adipose tissue (BAT) uncouples respiration, using lipids as an energy source while dissipating heat. Increases in BAT activity are protective against obesity, thus compounds that increase BAT activation may help prevent weight gain. Resveratrol (R) increases BAT activity by upregulating thermogenic genes. As phytochemicals have synergistic properties, our research strategy has included investigation of the efficacy of relatively low concentrations of phytochemical blends on BAT activation. Methods: Previously, we showed that R combined with genistein (G) and quercetin (Q) reduced weight gain in aged ovariectomized (OVX) female rats. In the current study, OVX rats were fed diets containing doses of phytochemicals with vitamin D (diet 1: 1000 mg/kg G; diet 2: 500 mg/kg G, 200 mg/kg R, and 1000 mg/kg Q; diet 3: 1000 mg/kg G, 400 mg/kg R, and 2000 mg/kg Q). Results: After 16 weeks, rats in the high dose group had a significantly smaller scapular BAT depot compared to non-OVX controls (0.74 g v 0.92 g;

Targeted Delivery of Antifungal Liposomes to Rhizopus delemar

Mucormycosis (a.k.a. zygomycosis) is an often-life-threatening disease caused by fungi from the ancient fungal division Mucoromycota. Globally, there are nearly a million people with the disease. Rhizopus spp., and R. delemar (R. oryzae, R. arrhizus) in particular, are responsible for most of the diagnosed cases. Pulmonary, rhino-orbito-cerebral, and invasive mucormycosis are most effectively treated with amphotericin B (AmB) and particularly with liposomal formulations (e.g., AmBisome®). However, even after antifungal therapy, there is still a 50% mortality rate. Hence, there is a critical need to improve therapeutics for mucormycosis. Targeting AmB-loaded liposomes (AmB-LLs) with the pathogen receptor Dectin-1 (DEC1-AmB-LLs) to the beta-glucans expressed on the surface of Aspergillus fumigatus and Candida albicans lowers the effective dose required to kill cells relative to untargeted AmB-LLs. Because Dectin-1 is an immune receptor for R. delemar infections and may bind it directly, we explored the Dectin-1-mediated delivery of liposomal AmB to R. delemar. DEC1-AmB-LLs bound 100- to 1000-fold more efficiently to the exopolysaccharide matrix of R. delemar germlings and mature hyphae relative to AmB-LLs. DEC1-AmB-LLs delivering sub-micromolar concentrations of AmB were an order of magnitude more efficient at inhibiting and/or killing R. delemar than AmB-LLs. Targeted antifungal drug-loaded liposomes have the potential to improve the treatment of mucormycosis

Enhanced effects of guggulsterone plus 1,25(OH)2D3 on 3T3-L1 adipocytes

Guggulsterone (GS) and 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] have been shown to influence adipogenesis in 3T3-L1 cells. We investigated the ability of GS and 1,25(OH)2D3, alone and in combination to inhibit adipogenesis and induce apoptosis in 3T3-L1 adipocytes. Maturing preadipocytes were treated with 1,25(OH)2D3 in combination with GS for 6 days during differentiation. GS and 1,25(OH)2D3 each inhibited lipid accumulation, but the combination potentiated the inhibition of lipid accumulation. Apoptosis was increased by 1,25(OH)2D3 while GS had no effect, but GS + 1,25(OH)2D3 increased apoptosis more than either compound alone. Furthermore, GS + 1,25(OH)2D3 caused a potentiated decrease in the expression of aP2 and farnesoid X receptor expression more than either compound alone. In addition, 1,25(OH)2D3 increased vitamin D receptor expression after 6 days, while GS had no effect. GS + 1,25(OH)2D3, however, caused a potentiated increase in the expression of VDR. These findings show that GS potentiates 1,25(OH)2D3\u27s anti-adipogenic and pro-apoptotic effects in maturing 3T3-L1 preadipocytes. © 2007 Elsevier Inc. All rights reserved

Resveratrol induces apoptosis and inhibits adipogenesis in 3T3-L1 adipocytes

Resveratrol, a phytoallexin, has recently been reported to slow aging by acting as a sirtuin activator. Resveratrol also has a wide range of pharmacological effects on adipocytes. In this study, we investigated the effects of resveratrol on adipogenesis and apoptosis using 3T3-L1 cells. In mature adipocytes, 100 and 200 μM resveratrol decreased cell viability dose-dependently by 23 ± 2.7%, and 75.3 ± 2.8% (p \u3c 0.0001), respectively, after 48 h treatment, and 100 μM resveratrol increased apoptosis by 76 ± 8.7% (p \u3c 0.0001). Resveratrol at 25 and 50 μM decreased lipid accumulation in maturing preadipocytes significantly by 43 ± 1.27% and 94.3 ± 0.3% (p \u3c 0.0001) and decreased cell viability by 25 ± 1.3% and 70.4 ± 1.6% (p \u3c 0.0001), respectively. In order to understand the anti-adipogenic effects of resveratrol, maturing 3T3-L1 preadipocytes were treated with 25 μM resveratrol and the change in the expression of several adipogenic transcription factors and enzymes was investigated using real-time RT-PCR. Resveratrol down-regulated the expression of PPARγ, C/EBPα, SREBP-1c, FAS, HSL, LPL and up-regulated the expression of genes regulating mitochondrial activity (SIRT3, UCP1 and Mfn2). These results indicate that resveratrol may alter fat mass by directly affecting cell viability and adipogenesis in maturing preadipocytes and inducing apoptosis in adipocytes and thus may have applications for the treatment of obesity. Copyright © 2008 John Wiley & Sons, Ltd

Octanoate and decanoate induce apoptosis in 3T3-L1 adipocytes

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. © Mary Ann Liebert, Inc

Enhanced pro-apoptotic and anti-adipogenic effects of genistein plus guggulsterone in 3T3-L1 adipocytes

Genistein (G), an isoflavone, and guggulsterone (GS), the active substance in guggulipid, have been reported to possess therapeutic effects for obesity. In the present study, we investigated the effects of combinations of G plus GS on apoptosis and adipogenesis in 3T3-L1 cells. In mature adipocytes, G and GS individually caused apoptosis, but combination of G plus GS significantly increased apoptosis, more than either compound alone. Furthermore, G plus GS caused a greater increase in procaspase-3 cleavage, Bax expression, cytochrome c release, and proteolytic cleavage of PARP than either compound alone. In maturing preadipocytes G and GS each suppressed lipid accumulation, but the combination potentiated the inhibition of lipid accumulation. These results suggest that combination of genistein and guggulsterone may exert anti-obesity effects by inhibiting adipogenesis and inducing apoptosis in adipocytes. © 2007 - IUBMB/IOS Press and the authors. All rights reserved

Anti-obesity effects of xanthohumol plus guggulsterone in 3T3-L1 adipocytes

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 μM and GS at 3.12 μM decreased lipid accumulation by 26 ± 4.5% (P \u3c .001) each, whereas XN1.5 + GS3.12 decreased lipid accumulation by 78.2 ± 1.8% (P \u3c .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. © 2009 Mary Ann Liebert, Inc. and Korean Society of Food Science and Nutrition

Withaferin A induces apoptosis and inhibits adipogenesis in 3T3-L1 adipocytes

Withaferin A (WA), a highly oxygenated steroidal lactone that is found in the medicinal plant Withania somnifera (also called ashwagandha) has been reported to have anti-tumor, anti-angiogenesis, and pro-apoptotic activity. We investigated the effects of WA on viability, apoptosis and adipogenesis in 3T3-L1 adipocytes. Pre- and post-confluent preadipocytes and mature adipocytes were treated with WA (1-25 μM) up to 24~hrs. Viability and apoptosis were measured by CellTiter-Blue® Cell Viability Assay and single strand DNA ELISA Assay, respectively. WA decreased viability and induced apoptosis in all stages of cells. Induction of apoptosis by WA in mature adipocytes was mediated by increased ERK1/2 phosphorylation and altered Bax and Bcl2 protein expression. The effect of WA on adipogenesis was examined by AdipoRed™ Assay after treating with WA (0.1-1 μM) during the differentiation period. WA decreased lipid accumulation in a dose-dependent manner and decreased the expression of peroxisome proliferator-activated receptor γ, CCAAT/enhancer binding protein α and adipocyte fatty acid binding protein. The effects on apoptosis and lipid accumulation were also confirmed with Hoechst staining and Oil Red O staining, respectively. These results show that WA acts on adipocytes to reduce cell viability and adipogenesis and also induce apoptosis. © 2008 IUBMB/IOS Press and the authors. All rights reserved

Enhanced effects of 1,25(OH) 2D 3 plus genistein on adipogenesis and apoptosis in 3T3-L1 adipocytes

Objective:To investigate the ability of 1,25(OH) 2D 3 (D) and genistein (G), alone and in combination, to inhibit adipogenesis and induce apoptosis in 3T3-L1 adipocytes.Methods and Procedures:3T3-L1 preadipocytes and mature adipocytes were incubated with various concentrations of D and G, alone and in combination, for 48 h. Viability was determined using the Cell Titer 96 Aqueous One Solution Cell Proliferation Assay. Post-confluent preadipocytes were incubated with D and G for up to 6 days during adipogenesis and lipid content was quantified by Nile Red dye; apoptosis was quantified by measurement of single-stranded DNA. Expression of adipocyte-specific proteins and VDR was analyzed by western blotting.Results:Combining D and G did not cause an enhanced effect on cell viability in either preadipocytes or mature adipocytes. In maturing preadipocytes, D at 0.5 nmol/l (D0.5) increased apoptosis by 47 ± 10.25 (P \u3c 0.05) and inhibited lipid accumulation by 28 ± 10 (P \u3c 0.001), while G at 25 mol/l (G25) had no significant effect. However, DG caused an enhanced apoptosis by 136 ± 12.6 (P \u3c 0.001) and enhanced inhibition of lipid accumulation by 82.46 ± 2.95 (P \u3c 0.001). Similarly, D0.5 alone decreased adipose-specific gene 422 (aP2) expression to 34.2 ± 2.3 and increased VDR expression levels by 41.8 ± 11 (P \u3c 0.001), but G25 showed no effect. However, D0.5G25 decreased aP2 expression to 52 ± 4.2 (P \u3c 0.05) and increased VDR expression levels by 131 ± 14.5 (P \u3c 0.0001).Discussion:These findings suggest that combining 1,25(OH) 2D 3 with genistein results in an enhanced inhibition of lipid accumulation and induction of apoptosis in maturing 3T3-L1 preadipocytes. © 2008 The Obesity Society