Synergistic Phytochemicals Fail to Protect Against Ovariectomy Induced Bone Loss in Rats

Suresh Ambati,Colette N. Miller,Erica F. Bass,Natalie M. Hohos,Diane L. Hartzell,Emily W. Kelso,Emily R. Trunnell,양정예,Mary Anne Della-Fera,Clifton A. Baile,Srujana Rayalam 한국식품영양과학회 2018 Journal of medicinal food Vol.21 No.10

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.

Combined Effects of Genistein, Quercetin, and Resveratrol in Human and 3T3-L1 Adipocytes

Hea Jin Park,Jeong-Yeh Yang,Suresh Ambati,Mary Anne Della-Fera,Dorothy B. Hausman,Srujana Rayalam,Clifton A. Baile 한국식품영양과학회 2008 Journal of medicinal food Vol.11 No.4

The natural compounds genistein (G), quercetin (Q), and resveratrol (R) have been reported to each exhibit anti-adipogenic activities in adipocytes and antiproliferative and pro-apoptotic activities in several cell types. We studied the combined effects of G, Q, and R on adipogenesis and apoptosis in primary human adipocytes (HAs) and 3T3-L1 murine adipocyte (MAs). Combined treatment with 6.25 μM G, 12.5 μM Q, and 12.5 μM R during the 14-day differentiation period caused an enhanced inhibition of lipid accumulation in maturing HAs that was greater than the responses to individual compounds and to the calculated additive response. Glycerol 3-phosphate dehydrogenase activity, a marker of late adipocyte differentiation, was decreased markedly in HAs treated with the combination of G+Q+R. In addition, combined treatment with 50 μM G, 100 μM Q, and 100 μM R for 3 days decreased cell viability and induced apoptosis in early- and mid- phase maturing and lipid-filled mature HAs. In contrast, no compound alone induced apoptosis. Oil Red O stain and Hoechst 33342 stain were performed to confirm the effects on lipid accumulation and apoptosis, respectively. We also determined whether MAs responded to the combination treatment similarly to HAs. As in HAs, G+Q+R treatment decreased lipid accumulation in maturing MAs and increased apoptosis in pre- and lipid-filled mature MAs more than the responses to G, Q, and R when used separately. These results show that lower concentrations of combined treatments with several natural compounds may be useful for treatments for obesity through the suppression of adipogenesis and enhanced adipocyte apoptosis.

Octanoate and Decanoate Induce Apoptosis in 3T3-L1 Adipocytes

Jeong-Yeh Yang,Srujana Rayalam,Mary Anne Della-Fera,Suresh Ambati,Clifton A. Baile 한국식품영양과학회 2009 Journal of medicinal food Vol.12 No.5

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.

Anti-Obesity Effects of Xanthohumol Plus Guggulsterone in 3T3-L1 Adipocytes

Rayalam, Srujana,Yang, Jeong-Yeh,Della-Fera, Mary Anne,Park, Hea-Jin,Ambati, Suresh,Baile, Clifton A. The Korean Society of Food Science and Nutrition 2009 Journal of medicinal food Vol.12 No.4

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 $1.5\;{\mu}M$ and GS at $3.12\;{\mu}M$ decreased lipid accumulation by $26\;{\pm}\;4.5%$ (P < .001) each, whereas XN1.5+GS3.12 decreased lipid accumulation by $78.2\;{\pm}\;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.

Preventing Bone Loss and Weight Gain with Combinations of Vitamin D and Phytochemicals

Ching-Yi Lai,Jeong-Yeh Yang,Srujana Rayalam,Mary Anne Della-Fera,Suresh Ambati,Richard D. Lewis,Mark W. Hamrick,Diane L. Hartzell,Clifton A. Baile 한국식품영양과학회 2011 Journal of medicinal food Vol.14 No.11

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.

Anti-Obesity Effects of Xanthohumol Plus Guggulsterone in 3T3-L1 Adipocytes

Srujana Rayalam,Jeong-Yeh Yang,Mary Anne Della-Fera,Hea Jin Park,Suresh Ambati,Clifton A. Baile 한국식품영양과학회 2009 Journal of medicinal food Vol.12 No.4

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 1.5μM and GS at 3.12μM 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.