IDENTIFICATION AND MECHANISM OF ANTI-OBESITY PHYTOCHEMICALS IN FOODS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 1007898
• Project Start Date: Oct 20, 2015
• Project End Date: Sep 30, 2020
• Program Code: [(N/A)]- (N/A)

Recipient Organization
RUTGERS, THE STATE UNIVERSITY OF NEW JERSEY
3 RUTGERS PLZA
NEW BRUNSWICK,NJ 08901-8559

Performing Department
Food Science

Non Technical Summary
The increasing prevalence of obesity is happening not only in the United States but also worldwide. According to WHO data, the prevalence of obesity has doubled since 1980. Almost 65% of the world's population lives in countries where overweight and obesity kills more people than underweight. Today, obesity affects not only adults but also children; 15-20% of children and adolescents are obese in the United States. Obesity is a serious h ealth problem worldwide. The primary cause of obesity is an energy imbalance. When energy intake is more than energy expenditure, the excess energy could be stored as triacylglycerol in the adipocytes. This excess energy can increase the number and size of adipocytes, eventually expanding the adipose tissue. Adipose tissue can be deposited under the skin and in the intra-abdominal area. More and more research has shown that adipose tissue is an endocrine organ that can regulate the whole-body energy homeostasis through the secretion of various adipose-derived hormones or adipokines and affects the function of other organs, such as the brain, liver, pancreas, and skeletal muscle. Therefore, obesity plays a key role in metabolic syndromes and is highly related to diabetes, cardiovascular disease, and cancer. The National Institutes of Health has developed some strategic plans for obesity research, including the discovery of biological mechanisms regulated to energy balance; understanding of the correlates, determinants, and consequences of obesity; the design and testing of interventions to promote healthy weight; the dissemination and implementation of research; and improvements in measurement tools, technology, and methods. Their ultimate goal is to prevent and treat obesity efficiently. Recently, researchers have been more interested in the role of natural compounds such as epigallocatechin-3-gallate from green tea, raspberry ketones from raspberries, and synephrine from orange in regulating lipid metabolism to decrease adipogenesis in vivo or in vitro.Our proposed research is seeking novel compounds from fruits and vegetables which have anti-adipogenic and anti-obesity activity and study their mode of actions.

Animal Health Component

20%

Research Effort Categories

Basic

70%

Applied

20%

Developmental

10%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
502	0999	1010	100%

Knowledge Area
502 - New and Improved Food Products;

Subject Of Investigation
0999 - Citrus, general/other;

Field Of Science
1010 - Nutrition and metabolism;

Keywords

anti-obesity

natural products

orange peel

functional foods

fruits

vegetables

spices

Goals / Objectives
Obesity is the leading health problem and is highly prevalent across the world. It is not only a simple disease but also contributes to the development of cardiovascular disease, type 2 diabetes, hypertension, fatty liver disease, cancer, and other pathological conditions. Genetic and environmental factors are both risk factors for obesity whereas environmental factors such as dietary behaviors and lifestyle are recognized to play predominant roles in causing this disease. Adipose tissue is an active endocrine and metabolic organ, which in addition to triacylglycerol (TG) storage, secretes various adipocytokines, cytokines, growth factors and hormones, implicating energy homeostasis and pathological processes. It is believed that combination of decreased caloric intake and physical exercise is effective for prevention of obesity, but it is difficult to have people to change lifestyle and control diet. Some anti-obesity drugs have been used to treat overweight or obese patients, however, with less efficacy and safety concern. Some literature shows that many phytochemicals and natural products are used to treat obesity as metabolic stimulates, appetite suppressants and starch blockers, and by regulation of glucose and lipid metabolism as well as targeting on adipocyte differentiation.The major objectives of this research are (1) to identify phytochemicals which have potential anti-obesity properties; (2) to understand the mechanism of inhibitory effect of polymethoxyflavones and other flavonoids on adipogenesis.

Project Methods
Bioassay directed fractionation and identification of natural products. We have extensive expertise in the isolation and structural identification of natural products from plants. Most natural products contain a complex mixture of compounds, and isolation begins with crude extracts. Complex mixtures of extracts from natural products will be fractionated using standard column chromatography. The bioactivity of the fractionated compounds will be established by appropriate bioassays. This process has come to be referred in the literature as "bioassay directed fractionation". Specific chemical fractionation will not be started until a fraction shows high activity in the bioassays. Once the bioactive fraction or fractions are identified, various fractionation techniques such as column chromatography, thin layer chromatography and high performance liquid chromatography will be used to isolate the pure compound, bioassays will continue to be used to guide the fractionation. Once the pure active compounds were obtained, they will be identified by HPLC/Mass spectrometry, tandem mass spectrometry, proton and C13 NMR spectrometry as well as various 2D NMR techniques, in some cases, X-ray crystallography will be needed to establish the absolute configuration of the molecules.Application of a cell based adipocyte differentiation and function assays for evaluation of the effect of test articles on adipocyte genes expression and functions We will use our already developed adipocyte differentiation and functional assays for bioactives anti-obesity related potential evaluation.2.1) Mouse 3T3-L1 preadipocytes purchased from the American Type Culture Collection (Rockville, MD, USA) were grown in DMEM supplemented with 2 mM glutamine (GIBCO BRL), 1% penicillin/streptomycin (10000 units of penicillin/mL and 10 mg streptomycin/mL), and 10% FCS at 37 °C under a humidified 5% CO2 atmosphere. Differentiation of 3T3-L1 preadipocytes performed as described before with a slightly modification. Briefly, cells were seeded into a 24-well (2 × 104/mL) or 10 cm dish with DMEM containing 10% FBS. Three days after full confluence (defined as day 0), cells were incubated in differentiation medium (DMI) containing 5 ?g/mL insulin, 0.5 mM IBMX, 1 ?M DEX and 2 ?M pioglitazone in DMEM containing 10% FBS for 2 days. The medium was then replaced by DMEM containing 10% FBS and 5 ?g/mL insulin with or without test compound, which was replaced every 2 days. The final concentrations of dimethyl sulfoxide (DMSO) in the culture medium were <0.05%. The cells were harvested after 8 days (at day 10) for Oil Red O staining and protein extraction.2.2) Cytotoxicity assay. The effect of test compound on cell viability of 3T3-L1 adipocyte was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). 3T3-L1 preadipocytes were seeded into a 96-well plate (2 × 104/mL) overnight and treated with various concentrations of test compound for 48 h. At the end of treatment, MTT (200 μg/mL) was added, and cells were incubated for another 4 h at 37 °C. After removal of the liquid, the MTT formazan were dissolved by DMSO. Cell viability was determined by scanning with an ELISA reader with a 570 nm filter.2.3) Oil Red O Staining. At the end of differentiation, cells were washed twice with phosphate-buffered saline (PBS), fixed with 10% formalin for 60 min, and stained with 0.5% Oil Red O in 2-propanol for 1 h at room temperature. Excess Oil Red O dye was washed twice with distilled water and then dried. The stained lipid droplets within were visualized by light microscope and photographed with a digital camera at 100× magnification. To quantify lipid accumulation, the stained lipid droplets were dissolved in 2-propanol, and the absorbance was measured at 520 nm.2.4) Western blot analysis. The total proteins of adipocytes and livers were extracted by addition of gold lysis buffer (50 mM Tris-HCl, pH 7.4; 1 mM NaF; 150 mM NaCl; 1 mM EGTA; 1 mM phenylmethanesulfonyl fluoride; 1% NP-40; and 10 ?g/mL leupeptin) to the cell pellets on ice for 30 min, followed by centrifugation at 10,000 ×g for 30 min at 4 oC. The total proteins were measured by Bio-Rad Protein Assay (Bio-Rad Laboratories, Munich, Germany). The samples (50 ?g of protein) were mixed with 5 × sample buffer containing 0.3 M Tris-HCl (pH 6.8), 25% 2-mercaptoethanol, 12% sodium dodecyl sulfate (SDS), 25 mM EDTA, 20% glycerol, and 0.1% bromophenol blue. The mixtures were boiled at 100 °C for 5 min and were subjected to 10% SDS-polyacrylamide minigels at a constant current of 20 mA. Electrophoresis was then carried out on SDS-polyacrylamide gels. Proteins on the gel were electrotransferred onto an immobile membrane (PVDF; Millipore Corp., Bedford, MA, USA) with transfer buffer composed of 25 mM Tris-HCl (pH8.9), 192 mM glycine, and 20% methanol. The membranes were blocked with blocking solution containing 20 mM Tris-HCl and then immunoblotted with primary antibodies and β-actin. The blots were rinsed three times with PBST buffer for 10 min each. Then blots were incubated with 1:5000 dilution of the horseradish peroxide (HRP)-conjugated secondary antibody (Zymed Laboratories, San Francisco, CA, USA) and then washed again three times with PBST buffer. The transferred proteins were visualized with an enhanced chemiluminescence detection kit (ECL; Amersham Pharmacia Biotech, Buckinghamshire, UK).

Progress 10/20/15 to 09/30/20

Outputs
Target Audience:This research ultimately targets consumers to give them guildance for the selection of right daily diet to reduce and prevent obesity, as well as to improve their health. The infomation we provided also assists plant breeders to produce foods with enhanced amounts of antiobesity beneficial compounds. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?One Masters student, Zisheng Han, has completed his Masters degree with a thesis entitled "LC-MS based metabolomics and sensory evaluation reveal the critical compounds of different grades of Huangshan Maofeng green tea' in October 2019.Z isheng is continuing his PhD degree with a project on tea and health How have the results been disseminated to communities of interest?Through peer-reviewed publications and conference presentations, we have made our research available to wide-range ofaudiences. What do you plan to do during the next reporting period to accomplish the goals?This is the end of this proposed project, but we will continuing to write few more articles and reviews about anti-obesity and potential phytochemicals which may manage the obesity.

Impacts
What was accomplished under these goals? The epidemic of obesity has become a worldwide public health concern in recent years, especially in the developed countries. In the United States, the obesity prevalence among adults has increased from 30.5% to 42.4% from 1999 to 2018(CDC, 2020). Obesity is considered as a central feature that increases risks for a cluster of metabolic syndrome (MetS),such as dyslipidemia, insulin resistance, hypertension, type 2 diabetes, fatty liver diseases, atherosclerosis, among others.(Hotamisligil, 2006). Given the prevalence of MetS in the modern life, the identification and exploration of dietary compounds against MetS is an attractive strategy for functional food development.Polymethoxyflavones (PMFs) are a unique class of flavonoids that have at least two methoxy groups on the flavone skeleton. PMFs are identified in the citrus seeds, leaves, stems, juice, and abundantly in the citrus peels. PMFs have demonstrated a broad spectrum of bioactivities, including neuroprotection, anti-inflammatory, anti-cancer, anti-obesity, anti-atherosclerosisactivities. Due to the good anti-inflammatory activity, PMFs have been investigated to protect against MetS for years.Compared to citrus flavanones (e.g. hesperidin and naringenin), PMFs show better bio-efficiency against dyslipidemia and obesity in vitro and in vivo. In recent five years, emerging studies find that PMFs can modulate the circadian clock and gut microbiota, besides regulating lipid- and glucose- metabolism in vivo. In addition, increasing attention has been paid for the regulation of metabolic homeostasis by the crosstalk of multiple organs, including the brain, pancreas, liver, adipose tissues, muscles and gastrointestinal (GI) track. Obesity is a serious public health issue. Since 1975, obesity in the population has tripled, increasing the risks of type 2 diabetes, cardiovascular diseases, and some cancers. The main cause of obesity is an energy imbalance between caloric intake and calories expended. Excessive energy is stored as triacylglycerol in the adipocytes, which are the main cells found in adipose tissue. There are two classic adipose adipocyte types: one is white adipocyte, which stores and releases energy,and the other is brown adipocyte, which burns fatty acids or glucose to produce heat for adaptive, non-shivering thermogenesis. Uncoupling protein 1 (UCP-1) is localized in the inner membrane of mitochondria and has an important role in this thermogenic activity. Specifically, UCP-1-expressing adipocytes develop in white adipose tissue (WAT) and perform thermogenic activity in response to certain stimuli referred to as beige or brite (brown in white) adipocytes. Therefore, beige fat could be a potential solution in the fight against obesity. Inguinal white adipose tissue in mice is enriched with beige adipocytes when it is stimulated by pharmacological or environmental factors, Hence, a WAT browning process could be developed. Recently, obesity and diet have been found to be highly associated with gut microbiota in mice and humans. Additionally, it is possible to brown fat by altering the composition of the gut microbiota or producing and transforming metabolites, such as short chain fatty acids, which could then be used as signaling molecules to induce browning. Many bioactive food components, especially those coming from fruits and vegetables, including berries, soybeans, spices, and citrus,have been shown to control the energy balance and thus improve body weight management. Soybeans (Glycine max) have been a traditional component in miso and doenjang in Asian countries for centuries, and soy isoflavones, including genistein,daidzein, and glycitein, are found in soybeans. Genistein is present in the greatest abundance, followed by daidzein, and both of these phytoestrogens have shown up in biological activities. 3'-hydroxydaidzein (OHD) is a metabolite of daidzein (DAI) that can be found in fermented soybean products, such as miso. DAI has been known to affect lipid accumulation, but the effect of OHD on lipid accumulation still needs to be investigated. In this study, we investigated the effects of OHD on mice with obesity induced by a high-fat diet. The results showed that mice treated with 0.1% OHD (HOHD) significantly reduced their body weight and inguinal fat without altering their food intake compared with the high-fat diet (HFD) group. The HOHD andDAI groups' hyperlipidemia was alleviated through decreased serum triglycerides and total cholesterol levels. The adipocyte sizes in inguinal fat were significantly smaller in the HOHD and DAI groups compared with the HFD group. Both the HOHDand DAI groups had increased PRDM16, C/EBP β, p-p38, SIRT1, PGC1 α, and UCP1 protein expression in their inguinal adipose tissue compared with the HFD group. Moreover, the OHD and DAI groups had significantly lower amounts of Lachnospira and GCA_900066225 compared with the HFD group. Collectively, 3'-hydroxydaidzein can ameliorate obesityinduced by a high-fat diet by stimulating the browning of white adipose tissue and modulating gut microbiota. In another study, the aged citrus peels (chenpi) have been used as dietary supplement for gastrointestinal health maintenance in China, and recently reported to exhibit anti-obesity activity. However, the relationship between the modulation effect of chenpi on gut microbiota and obesity prevention is not clearly understood. In this study, mice were fed with high-fat diet (HFD), HFD supplemented with 0.25%- and 0.5%- chenpi extract, and normal diet for 11 weeks. Chenpi extract significantly increased fecal short chain fatty acids by 43% for acetic acid, and 86% for propionic acid. In addition, chenpicould decrease Proteobacteria prevalence and the ratio of Firmicutes to Bacteroidetes by about 88% and 70%, respectively.Moreover, this study was the first work to demonstrate the dynamics of two beneficial bacteria -- Akkermansia spp. andAllobaculum spp. in the dose- and time-dependent manners of chenpi treatment via monitoring the dynamical change of the gut microbiota. The metagenomics analysis of the gut microbiota showed that several pathways were enhanced dynamically,such as two-component systems, tight junctions, staphylococcus aureus infections and others. The improved biological process of metabolism especially on benzoate-derivatives might refer to the increased metabolic transformation of polymethoxyflavones from chenpi in the colon. Our study indicated that the modulation effect of chenpi on the gut microbiota was an important pathway for its anti-obesity mechanisms.

Publications

• Type: Journal Articles Status: Published Year Published: 2019 Citation: Yang, G.; Lin, C.C.; Yang, Y.; Yuan, L.; Wang, P.; Wen, X.; Pan, M.H.; Zhao, H.; Ho, C.T.; Li, S. Nobiletin prevents TMAO-induced vascular inflammation via inhibition of the NF-kB/MAPK pathways. Journal of Agricultural and Food Chemistry,2019, 67(22), 6169-6176.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Lu,K.H.; Lee, H.Y.; Chu, Y.L.; Ho, C.T.; Sheen, L.Y. Bitter orange peel extract induces endoplasmic reticulum-mediatedautophagy in human hepatoma cells. Journal of Functional Foods, 2019, 60, 103404.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Wu, W.K.; Chen, C.C., Liu, P.Y.; Panyod, S.; Liao, B.Y.; Chen, P.c.; Kao, H.L.; Kuo, H.Ch.; Kuo, C.H.; Chiu, T.H.T.; Chen,R.A.; Chuang, H.L.; Huang, Y.T.; Zou, H.B.; Hsu, C.C.; Chang, T.Y.; Lin, C.L.; Ho, C.T.; Yu, H.T.; Sheen, L.Y.; Wu, M.S.Identification of TMAO-producer phenotype and host-diet-gut dysbiosis by carnitine challenge test in human and germ-freemice. Gut, 2019, 68(8), 1439-1449.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Chen, P.Y.; Li, S.; Koh, Y.C.; Wu, J.C.; Yang, M.J.; Ho, C.T.; Pan, M.H. Oolong tea extract and citrus peel polymethoxyflavones reduce transformation of L-carnitine to trimethylamine-N-oxide and decrease vascular inflammationin L-carnitine feeding mice. Journal of Agricultural and Food Chemistry, 2019, 67(28), 7869-7879.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Guo, T.; Song, D.; Ho, C.T.; Zhang, X.; Zhang, C.; Cao, J.; Wu, Z. Omics analyses of gut microbiota in a circadian rhythm disorder mouse model fed with oolong tea polyphenols. Journal of Agricultural and Food Chemistry, 2019, 67(32), 8847-8854.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Zhang, L.; Ho, C.T.; Zhou, J.; Santos, J.S.; Armstrong, L.; Granato, D. Chemistry and biological activities of processed Camellia sinensis teas: A comprehensive review. Comprehensive Reviews in Food Science and Food Safety, 2019, 18(5),1474-1495.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Koh, Y.C.; Li, S.; Chen, P.Y.; Wu, J.C.; Kalyanam, N.; Ho, C.T.; Pan, M.H. Prevention of vascular inflammation bypterostilbene via trimethylamine-N-oxide reduction and mechanism of microbiota regulation. Molecular Nutrition & Food Research, 2019, 63(20), 1900514
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Guo, T.; Ho, C.T.; Zhang, X.; Cao, J.; Wang, H.; Shao, X.; Pan, D.; Wu, Z. Oolong tea polyphenols ameliorate circadian rhythm of intestinal microbiome and liver clock genes in mouse model. Journal of Agricultural and Food Chemistry, 2019,67(43), 11969-11976.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Pan, M.H.; Koh, Y.C.; Lee, T.L.; Wang, B.; Chen, W.K.; Nagabhushanam, K.; Ho, C.T. Resveratrol and oxyresveratrol activate thermogenesis via different transcriptional co-activators in high-fat diet-induced obesity in mice. Journal of Agricultural and Food Chemistry, 2019, 67(49), 13605-13616.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Pan, M.H.; Li, M.Y.; Tsai, M.L.; Pan, C.Y.; Badmaev, V.; Ho, C.T.; Lai, C.S.. A mixture of citrus polymethoxyflavones,green tea polyphenols and lychee extracts attenuates adipogenesis in 3T3-L1 adipocytes and obesity-induced adipose inflammation in mice. Food & Function, 2019, 10(12), 7667-7677.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Chou, Y.C.; Li, S.; Ho, C.T.; Pan, M.H. Preparation and evaluation of self-microemulsifying deliver system containing 5-demethyltangeretin on inhibiting xenograft tumor growth in mice. International Journal of Pharmaceutics, 2020, 579,119134.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Hung, W.L.; Ho, C.T.; Pan, M.H. Targeting the NLRP3 inflammasome in neuroinflammation: Health promoting effects of dietary phytochemicals in neurological disorders. Molecular Nutrition & Food Research, 2020, 64(4), 1900550.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Guo, S.; Ren, X.; He, K.; Chen, X.; Zhang, S.; Roller, M.; Zheng, B.; Zheng, Q.; Ho, C.T.; Bai, N. The anti-diabetic effect of eight Lagerstroemia speciosa leaf extracts based on the contents of ellagitannins and ellagic acid derivatives. Food &Function, 2020, 11(2), 1560-1571.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Zhang, M.; Zhu, J.; Zhang, X.; Zhao, D.g.; Ma, Y.; Li, D.; Ho, C.T.; Huang, Q. Aged citrus peel (Chenpi) extract causes dynamic alternation of colonic microbiota in high-fat diet induced obese mice. Food & Function, 2020, 11(3), 2667-2678.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Lu, M.; Chen, C.; Lan, Y.; Xiao, J.; Li, R.; Huang, J.; Huang, Q.; Cao, Y.; Ho, C.T. Capsaicin-the major bioactive ingredient of chili peppers: Bio-efficacy and delivery systems. Food & Function, 2020, 11(4), 2848-2860.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Lee, P.S.; Chiou, Y.S.; Nagabhushanam, K.; Ho, C.T.; Pan, M.H. 3-Hydroxypterostilbene potentially alleviates obesity exacerbated colitis in mice. Journal of Agricultural and Food Chemistry, 2020, 68(19), 5365-5374.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Feng, K.; Lan, Y.; Zhu, X.; Li, J.; Chen, T.; Huang, Q.; Ho, C.T.; Chen, Y.; Cao, Y. Hepatic lipidomics analysis reveals the antiobesity and cholesterol-lowering effects of tangeretin in high-fat diet-fed rats. Journal of Agricultural and Food Chemistry, 2020, 68(22), 6142-6153
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Yan, R.; Ho, C.T.; Zhang, X. Interaction between tea polyphenols and intestinal microbiota in host metabolic diseases from the perspective of the gut-brain axis. Molecular Nutrition & Food Research, 2020, 64(14), 2000187.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Li, R.; Lan, Y.; Chen, C.; Cao, Y.; Huang, Q.; Ho, C.T.; Lu, M. Anti-obesity effects of capsaicin and the underlying mechanism: a review. Food & Function, 2020, 11(9), 7356-7370.
• Type: Journal Articles Status: Published Year Published: 2020 Citation: Zhang, M.; Xin, Y.; Feng, K.; Yin, B.; Kan, Q.; Xiao, J.; Cao, Y.; Ho, C.T.; Huang, Q. Comparative analyses of bioavailability, biotransformation, and excretion of nobiletin in lean and obese rats. Journal of Agricultural and FoodChemistry, 2020, 68(39), 10709-10718

Progress 10/01/18 to 09/30/19

Outputs
Target Audience:This project targets consumers to give them guildance for the selection of right daily diet to reduce and prevent obesity, as well as improve their health. The infomation we provided will also assist plant breeders to produce foods with enhanced amounts of antiobesity beneficial compounds. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?One PhD student, Siyu Wang has obtained her PhD degree with thesis entitled 'Hydroxycinnamic acid amides from Lycium barbarum and their anti-inflammatory mechanisms" in May 2018. She is continuing doing postoctoral training in University of Pennsylvania. How have the results been disseminated to communities of interest?Through peer-reviewed publications and conference presentations, we have made our research well-publicized. What do you plan to do during the next reporting period to accomplish the goals?We are planning to publish several research papers in jounals such as Journal of Agricultural and Food Chemistry. A paper on the anti-obesity effect of polymethoxyflavones will be presented at the International Conference on Food Factor held in Kobe, Japan in December 2019.

Impacts
What was accomplished under these goals? Obesity is characterized as a surplus of energy from consumption that exceeds energy expenditure and is stored as fat. According to the previous study, more than 60% of deaths are related to high body mass index (BMI), and the prevalence of obesity may reach nearly 40% of adults around the globe (18% in men and over 21% in women) in the year 2025. Obesity is normally defined as a BMI of ≥30 kg/m2. However, BMI does not provide much information on body fat distribution. As mentioned, obesity is positively related to deadly diseases because it is a debilitating health condition that is associated with metabolic syndrome, and it develops into several adverse comorbid diseases including cardiovascular disease, type II diabetes and the exacerbation of cancer. Therefore, addressing the obesity issue is imperative in order to abate economic and medical burdens in coming years. Fat tissue can be divided into two categories according to its structure and function. White adipose tissue (WAT) regulates systemic homeostasis and stores energy as triacylglycerols (TG) while brown adipose tissue (BAT) can expend lipids via thermogenesis. Heat generation via stored calorie consumption may affect metabolic outcomes and thermogenesis in BAT is reported to be negatively correlated to adiposity, blood glucose and insulin resistance. Therefore, there is elevated interest in the relationship between thermogenesis and metabolic-related diseases. In most animals, BAT can be identified by a relatively small lipid droplet in terms of capacity, due to the large number of mitochondria. It is responsible for non-shivering thermogenesis, which is known as adaptive thermogenesis. Brown-in-white (BRITE), also known as beige adipocyte, is a sub-type of adipose tissue that arises from WAT and promotes energy burning, which usually happens upon adrenergic stimulation. Because brown and/or beige adipose tissue has an inverse correlation with obesity, a hypothesis postulates that signaling pathways activating BAT thermogenesis and the beiging of WAT could be strategies to attenuate the condition of obesity. Notably, some phytochemicals or plant extracts have exerted the capability to activate fat browning in previous studies, including polyphenolic extract from green tea leaves, genistein, cyanidin-3-glucoside and immature Citrus reticulate extract containing nobiletin and tangeretin. Thermogenesis is a later concept in obesity prevention and aside from that, anti-adipogenesis, anti-lipogenesis and inducing lipolysis and β-oxidation are much more common strategies. During β-oxidation, fatty acids from broken triacylglycerols during lipolysis are oxidized, and two carbon units are removed from acyl chain into acetyl-CoA in mitochondria to be involved in Krebs cycle. Some phytochemicals, such as forskolin, exhibit anti-obesity capacity by stimulating β-oxidation. Therefore, diet-induced lipolysis/β-oxidation and thermogenesis can be effectual chemopreventive action. Resveratrol and oxyresveratrol are both natural stilbenoids that have been reported for their anti-obesity efficacy. However, the comparison between the two as they relate to thermogenesis as well as the differences in their underlying mechanisms are still not widely discussed. Our result reveals that both resveratrol and oxyresveratrol could elevate the expression of thermogenesis-related protein expression including UCP1 (Uncoupling protein-1) and PRDM (PR domain containing 16) via Sirt1/PGC-1α (Sirtuin 1/Peroxisome proliferation gamma coactivator-1 α) activation. However, it is suggested that the transcriptional factor PPARα (Peroxisome proliferator-activator receptor α) was activated by resveratrol (1.38 ± 0.07 fold) but not oxyresveratrol. Conversely, C/EBPβ (CCAAT/Enhancer-binding protein β) was upregulated by oxyresveratrol (1.58 ± 0.05 fold) but not by resveratrol. On the other hand, CPT1 (Carnitine palmitoyltransferase) was found to be significantly activated at lower concentrations of oxyresveratrol up to 1.89 ± 0.04 fold as compared to HFD, and it could be a leading reason for UCP1 activation. Lastly, adiponectin expression was promoted in all experimental groups (1.53 ± 0.08 and 1.49 ± 0.11 fold in RES and HOXY, respectively), which could be an activator for mitochondrial biosynthesis and UCP1 expression.

Publications

• Type: Journal Articles Status: Published Year Published: 2018 Citation: Tung, Y.C.; Hsieh, T.H.; Yang, M.J.; Hung, W.L.; Ho, C.T.; Pan, M.H. The effects of the extracts of oolong tea and its metabolites from Andraca theae in high fat diet induced obese Wistar rat. Food Science and Human Wellness 2018, 7(2), 120-124.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Sung, J.; Ho, C.T.; Wang, Y. Preventive mechanism of bioactive dietary foods on obesity-related inflammation and diseases. Food & Function 2018, 9(12), 6081-6095.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Lee, P.S.; Teng, C.Y.; Kalyanam, N.; Ho, C.T.; Pan, M.H. Garcinol reduces obesity in high-fat-diet-fed mice by modulating gut microbiota composition. Molecular Nutrition & Food Research 2019, 63(2), 1800390.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Feng, K.; Zhu, X.; Chen, T.; Peng, B.; Lu, M.; Zheng, H.; Huang, Q.; Ho, C.T.; Chen, Y.; Cao, Y. Prevention of obesity and hyperlipidemia by heptamethoxyflavone in high-fat diet-induced rats. Journal of Agricultural and Food Chemistry 2019, 67(9), 2476-2489.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Wang, S.; Pan, M.H.; Hung, W.L.; Tung, Y.C.; Ho, C.T. From white to beige adipocytes: therapeutic potential of dietary molecules against obesity and their molecular mechanisms. Food & Function 2019, 10(3), 1263-1279
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Meng, Q.; Li, S.; Huang, J.; Wei, C.C.; Wan, X.; Sang, S.; Ho, C.T. Importance of the nucleophic property of tea polyphenols. Journal of Agricultural and Food Chemistry 2019, 67(19), 5379-5383.

Progress 10/01/17 to 09/30/18

Outputs
Target Audience:The project will target consumers to help them to select daily diets to reduce obesity and obesity-associated diseases such as diabetes, inflammation and cadiovascular diseases, and improve their health. We will also help plant breeders to produce foods with enhanced amounts of anti-obesity phytochemicals. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?We have used this project to train two visiting scholars, one of them, Professor Qing Meng from Sichuan, China, and the other, Dr. Chia-Cheng Wei from Taiwan. Both of them were doing a good job. Dr. Meng returned to China and continue the research on the anti-obesity of tea products; and Dr. Wei took an Assistant Professor position at National Taiwan University in Taiwan, and will also work on anti-obesity. How have the results been disseminated to communities of interest?We presented papers in ACS annual meetings in Boston, August 2018. We also presented a paper in the 2018 Annual Meeting of Nutraceuticals and Functional Foods in Canada. What do you plan to do during the next reporting period to accomplish the goals?We will continue our study onthe anti-obesity of phytochemicals from citrus peels and pay particular attention on the role of gut microbiota plays when they interact with phytochemicals and the effects on anti-obesity

Impacts
What was accomplished under these goals? Obesity has become a major health problem worldwide and is associated with low-grade chronic inflammation and intestinal dysbiosis. The gut microbiota is a key player in the pathophysiology of diet-induced obesity. This study was conducted to investigate the chemopreventive effects of garcinol, a polyisoprenylated benzophenone derivative isolated from the fruit rind of Garcinia indica. We sought to delineate how garcinol protects against obesity in high-fat diet (HFD)-induced mice, and determined whether its anti-obesity effects are related to the gut microbiota. The results showed that garcinol reduced HFD-fed mice body weight gain and relative visceral adipose tissue fat weight in a dose-dependent manner. Furthermore, garcinol markedly reduced the serum levels of glutamate pyruvate transaminase, total cholesterol, and triacylglycerol. The 16S rRNA gene sequence data indicated that garcinol not only reversed HFD-induced gut dysbiosis--as indicated by the decreased Firmicutes-to-Bacteroidetes ratios but also controled inflammation by increasing intestinal probiotics, Akkermansia spp. The increased Akkermansia spp. population also affected the AMP-activated protein kinase α signaling pathway in adipocytes adipogenesis. Since promoting browning of white adipose tissue (WAT) helps to maintain energy homeostasis. Previous studies have found that citrus fruit exhibits a number of biological activities. Although most citrus fruit drop has been considered agricultural waste, the ability to use it may be desirable. In another project, we investigate the anti-obesity effects of immature citrus fruits in high-fat diet (HFD)-fed mice. The main phytochemical components of immature Citrus reticulatain water extraction analyzed by HPLC are synephrine, narirutin, hesperidin, nobiletin, and tangeretin (16.0±1.08, 4.52±0.31, 9.14±0.32, 2.54±0.07, 1.67±0.05 mg/g, respectively). Oral administration of 1% Immature Citrus ReticulataExtract (ICRE) for 11 weeks markedly reduced body weight gain, epididymal fat weight, fasting blood glucose, serum triglyceride, and total cholesterol (P< 0.05 for all). In addition, histological analysis revealed that dietary ICRE decreased adipocyte size and hepatic steatosis compared to the HFD group (P< 0.05 for both). Furthermore, we found that mice treated with ICRE have improved cold tolerance during acute cold challenge. These effects were associated with increased expression of uncoupling protein 1 (UCP1) and thermogenic genes in inguinal WAT. Taken together, these results suggest that ICRE can prevent obesity and lipid accumulation through induction of brown-like adipocyte formation.

Publications

• Type: Journal Articles Status: Published Year Published: 2018 Citation: Zhang, X.; Chen, Y.; Zhu, J.; Zhang, M.; Ho, C.-T.; Huang, Q.; Cao, J. Metagenomics analysis of gut microbiota in a high fat diet-induced obesity mouse model fed with (-)-epigallocatechin 3-O-(3-O-methyl) gallate (EGCG3?Me). Molecular Nutrition & Food Research 2018, 62(13), 1800274.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Kou, X.; Wang, X.; Ji, R.; Liu, L.; Qiao, Y.; Lou, Z.; Ma, C.; Li, S.; Wang, H.; Ho, C.-T. Occurrence, biological activity and metabolism of 6-shogaol. Food & Function 2018, 9(3), 1310-1327.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Huang, J.; Wang, X.; Tao, G.; Song, Y.; Ho, C.-T.; Zheng, J.; Ou, S. Feruloylated oligosaccharides from maize bran alleviate the symptom of diabetes in streptozotocin-induced type 2 diabetic rats. Food & Function 2018, 9(3), 1779-1789.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Wang, S.; Suh, J. H.; Hung, W. L.; Zheng, X.; Wang, Y.; Ho, C.-T. Use of UHPLC-TripleQ with synthetic standards to profile anti-inflammatory hydroxycinnamic acid amides in root barks and leaves of Lycium barbarum. Journal of Food and Drug Analysis 2018, 26(2), 578-582.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Hung, W. L.; Chang, W. S.; Lu, W. C.; Wei, G. J.; Wang, Y.; Ho, C. T.; Hwang, L. S. Pharmacokinetics, bioavailability, tissue distribution and excretion of tangeretin in rat. Journal of Food and Drug Analysis 2018, 26(2), 849-857.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Xu, N.; Chu, J.; Wang, M.; Chen, L.; Zhang, L.; Xie, Z.; Zhang, J.; Ho, C.-T.; Li, D.; Wan, X. Large yellow tea attenuates macrophage-related chronic inflammation and metabolic syndrome in high-fat diet treated mice. Journal of Agricultural and Food Chemistry 2018, 66, 66(15), 3823-3832.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Hsu, B. Y.; Pan, S. Y.; Wu, L. Y.; Ho, C.-T.; Hwang, L. S. Hypoglycemic activity of Chenopodium formosanum Koidz. components using a glucose uptake assay with 3T3-L1 adipocytes. Food Bioscience 2018, 24, 9-16.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Zhang, X.; Zhang, M.; Ho, C.-T.; Guo, X.; Wu, Z.; Weng, P.; Yan, M.; Cao, J. Metagenomics analysis of gut microbiota modulatory effect of green tea polyphenols by high fat diet-induced obesity mice model. Journal of Functional Foods 2018, 46, 268-277.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Chou, Y. C.; Ho, C.-T.; Pan, M. H. Stilbenes: Chemistry and molecular mechanisms of anti-obesity. Current Pharmacology Reports 2018, 4(3), 202-209.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Tung, Y. C.; Chang, W. T.; Li, S.; Wu, C. C.; Badmaev, V.; Ho, C.-T.; Pan, M. H. Citrus peel extracts attenuated obesity and modulate gut microbiota in mice with high-fat diet-induced obesity. Food & Function 2018, 9(6), 3363-3373.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Chen, J. W.; Kong, Z. L.; Tsai, M. L.; Lo, C. Y.; Ho, C.-T.; Lai, C. S. Tetrahydrocurcumin ameliorates free fatty acid-induced hepatic steatosis and improves insulin resistance in HepG2 cells. Journal of Food and Drug Analysis 2018, 26(3), 1075-1085.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Chou, Y. C.; Ho, C.-T.; Pan, M. H. Immature Citrus reticulata extract promotes browning of beige adipocytes in high-fat diet-induced C57BL/6 mice. Journal of Agricultural and Food Chemistry 2018, 66(37), 9697-9703.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Lu, M.; Cao, Y.; Xiao, J.; Song, M.; Ho, C.-T. Molecular mechanisms of the anti-obesity effect of bioactive ingredients in common spices: A review. Food & Function 2018, 9(9), 4569-4581.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Gosslau, A.; Zachariah, E.; Li, S.; Ho, C.-T. Anti-diabetic effects of a theaflavin-enriched black tea extract in the obese ZDF rat model. Journal of Food Bioactives 2018, 3, 151-160.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Suh, J. H.; Wang, Y.; Ho, C.-T. Natural dietary products and their effects on appetite control. Journal of Agricultural and Food Chemistry 2018, 66, 36-39.
• Type: Journal Articles Status: Published Year Published: 2018 Citation: Pan, M. H.; Wu, J. C.; Ho, C.-T.; Lai C. S. Antiobesity molecular mechanisms of action: Resveratrol and pterostilbene. BioFactors 2018, 44(1), 50-60.

Progress 10/01/16 to 09/30/17

Outputs
Target Audience:This project will target consumers to help them to select daily diets to reduce obesity and improve their health. We will also help plant breeders to produce foods with enhanced amounts of anti-obesity beneficial compounds. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?One PhD student, Muwen Lu has obtained her PhD degree with thesis entitled "Delivery of nutraceuticals using novel processing methods and emulsion-based formulations with enhanced dissolution, bioaccessibility and bioavailability" in May 2017. Dr. Mu has returned to China and started her Associate Professor job in Southern China Agricultural University in Guangzhou, China. How have the results been disseminated to communities of interest?Through peer reviewed publications and conference presentations. More than 100 colleagues were in attendance for the ACS National Meeting presentation on the health effect of omega-3 fatty acids and fish oil. What do you plan to do during the next reporting period to accomplish the goals?We are planning to publish several research papers in journals such as Journal of Agricultural and Food Chemistry, Food and Function etc. I will organize a symposium on "Chemistry, Flavor and Health Effects of Tea" for the American Chemical Society annual meeting in Boston in August 2018.

Impacts
What was accomplished under these goals? The prevalence of overweight and obesity has increased substantially worldwide over the past decade. It is simply defined as a condition by abnormal or excessive fat accumulation in adipose tissue resulted from imbalance between energy intake and expenditure. Nowadays, obesity has been recognized with huge impact on human health because it per se is not only regarded as a simple disease but also highly contributes to the development of the metabolic diseases and cardiovascular diseases. The effective obesity prevention and treatment is known as dietary control- and physical exercise-based behavioral therapies and/or in combination of pharmacologic therapies, however, it is difficult for most people because of their lifestyle. Although many pharmacological agents are current available for treatment of obesity but with limited efficacy and concerned side effects. Thus, research in identification of natural products from diet and herbs with anti-obesity properties is increasing popular recently. Flavonoids are naturally polyphenolic compounds that widely occurring almost in fruits, vegetables, herbs, other plants and also known as essential nutrients for human because they cannot be biosynthesized in animals. It was suggested that flavonols, flavones and isoflavones are the most abundant in human diet, such as tea, fruits and soybean. The physiological, biological properties and beneficial effects for human health of flavonoids are extensively investigated. Citrus peel has been originally used as traditional Chinese medicine for numerous purposes since ancient times. Polymethoxyflavones (PMFs) are a unique class of flavonoids that with two or more methoxy groups and almost exclusively in citrus peel. They are reported to display a broad spectrum of biological and pharmacological properties including anti-obesity. Several in vitro and in vivo studies show that PMFs or hydroxylated PMFs (characterized by the presence of one or two hydroxyl groups instead of methoxy groups) inhibit adipogenesis in adipocytes and reduce adipose mass in diet-induced obese mice. Green tea (Camellia sinensis) is regarded as nutraceuticals of modern life because of its health-promoting and disease prevention effect. The beneficial effect of green tea against obesity has been recognized and associated with the rich in polyphenols, in particular catechins such as epigallocatechin-3-gallate (EGCG). Consumption of green tea polyphenols reduces body weight and fat mass in animal models as well as several clinical trials through multiple mechanisms including increased energy expenditure, fat oxidation, suppression of adipocyte differentiation and proliferation and inhibition of fat absorption. Lychee (Litchi chinensis Sonnerat) is a subtropical fruit that contains abundant phenolic compounds, such as flavanol, and exhibits biological activities. Previous studies indicate that lychee fruit extract contains oligomerized flavanols. Various in vitro and in vivo studies also demonstrated numerous biological activities of lychee fruit extract with oligomerized flavanols, including antioxidant, anticancer, antiviral [20], anti-inflammatory, and improving insulin resistance. Dietary lychee fruit extract with oligomerized flavanols reduced adipose mass, adipocyte size and adipokines secretion as well as hepatic steatosis in high fat diet-fed mice through modulating multiple molecules such as PPARγ, mTOR/SREBP-1 and AMPK signaling. SlimTrym, is a botanical formula including citrus PMFs, green tea extract and lychee extract. The citrus peel extract in the SimTrym formula is standardized for 65% of total PMFs and the major PMFs are nobiletin and tangeretin with 24.0% and 8%, respectively. The green tea extract contains 70% of green tea polyphenols and 26% of catechins. The Lychee extract contains oligomerized flavanols. No reports are available on the weight-loss effect of combination of green tea polyphenols, lychee extract, and pcitrus PMFs. Male C57BL/6 mice were fed a normal diet (ND), high fat diet (HFD) or HFD containing 0.1% or 0.5% SlimTrym for sixteen weeks. Dietary SlimTrym significantly reduced weight gain and relative perigonadal, retroperitoneal, mesenteric fat weight as well as the size of adipocyte in HFD-fed mice. SlimTrym supplementation also effectively diminished hepatic steatosis and the serum levels of glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT), triacylglycerol (TG) and total cholesterol (TCHO). Down-regulation of peroxisome proliferator-activated receptor (PPAR)γ, sterol regulatory element-binding protein (SREBP)-1, and the activation of AMP-activated protein kinase (AMPK) signaling by SlimTrym in both adipose tissue and liver may be responsible for the observed anti-obesity effects. SlimTrym supplementation potentially diminished diet-induced obesity and hepatic steatosis via regulating AMPK signaling and molecules involved in lipid metabolism.

Publications

• Type: Journal Articles Status: Published Year Published: 2016 Citation: Tung, Y.C.; Li, S.; Huang, Q.; Hung, W.L.; Ho, C.T.; Wei, G.J.; Pan, M.H. 5-Demethylnobiletin and 5-acetoxy-6,7,8,3�,4�-pentamethoxyflavone suppress lipid accumulation by activating the LKB1-AMPK pathway in 3T3-L1 preadipocytes and high-fat diet-fed C57BL/6 mice. Journal of Agricultural and Food Chemistry 2016, 64(16), 3196-3205.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Wang, Y.; Lee, P.S.; Chen, Y.F.; Ho, C.T.; Pan, M.H. Suppression of adipogenesis by 5-hydroxy-3,6,7,8,3�,4�-hexamethoxyflavone from orange peel in 3T3-L1 cells. Journal of Medicinal Food 2016, 19(9), 830-835.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Tung, Y.C.; Lin, Y.H.; Chen, H.J.; Chou, S.C.; Cheng, A.C.; Kalyanam, N.; Ho, C.T.; Pan, M.H. Piceatannol exerts anti-obesity effects in C57BL/6 mice through modulating adipogenic proteins and gut microbiota. Molecules 2016, 21(11), 1419/1-1419/15.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Pan, M.H.; Tung, Y.C.; Yang, G.; Li, S.; Ho, C.T. Molecular mechanisms of the anti-obesity effect of bioactive compounds in tea and coffee. Food & Function 2016, 7(11), 4481-4491.
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Tung, Y.C.; Hsieh, P.H.; Pan, M.H.; Ho, C.T. Cellular models for the evaluation of antiobesity effect of selected phytochemicals from food and herbs. Journal of Food and Drug Analysis 2017, 25(1), 100-110.
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Tang, W.; Li, S.; Liu, Y.; Huang, M.T.; Ho, C.T. Anti-inflammatory effects of trans-2,3,5,4?-tetrahydroxystilbene 2-O-?-glucopyranoside (THSG) from Polygonum multiflorum (PM) and hypoglycemic effect of cis-THSG enriched PM extract. Journal of Functional Foods 2017, 34, 1-6.
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Guo, J.; Cao, Y.; Ho, C.T.; Jin, S.; Huang, Q. Aged citrus peel (chenpi) extracts reduces lipogenesis in differentiating 3T3-L1 adipocytes. Journal of Functional Foods 2017, 34, 297-303.
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Lu, M.; Cao, Y.; Ho, C.T.; Huang, Q. The enhanced anti-obesity effect and reduced gastric mucosa irritation of capsaicin-loaded nanoemulsions. Food & Function 2017, 8(5), 1803-1809.
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Lai, C.S.; Wu, J.C.; Ho, C.T.; Pan, M.H. Chemoprevention of obesity by dietary natural compounds targeting mitochondrial regulation. Molecular Nutrition & Food Research 2017, 61(6), 1600721.
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Pan, M.H.; Yang, G.; Li, S.; Li, M.Y.; Tsai, M.L.; Wu, J.C.; Badmaev, V.; Ho, C.T.; Lai, C.S. Combination of citrus polymethoxyflavones, green tea polyphenols and lychee extracts suppresses obesity and hepatic steatosis in high-fat diet induced obese mice. Molecular Nutrition & Food Research 2017, 61(11), 1601104.
• Type: Conference Papers and Presentations Status: Other Year Published: 2017 Citation: Ho, C.T., Wang, Y. Reactive carbonlyl species from the oxidation of omega-3 and omega-6 fatty acids and method for their intervention. Paper presented at the 254th American Chemical Society National Meeting, Washington, DC, August 2017.
• Type: Conference Papers and Presentations Status: Other Year Published: 2017 Citation: Wang, Y. J. Hsiao, T-Y , Li, S., Pan, M.H., Ho, C.T., Lo, C. Inhibitory effect of black tea theaflavins on advanced glycation end product formation in the fructose-induced protein system. Paper presented at the 254th American Chemical Society National Meeting, Washington, DC, August 2017.
• Type: Conference Papers and Presentations Status: Other Year Published: 2017 Citation: Wu, W., Zhao, Y., Sang, S., Ho, C.T. Tetrahydroisoquinoline derivatives by reaction of dopamine with methylglyoxal: Potential neurotoxins associated with Parkinsons disease. Paper presented at the 254th American Chemical Society National Meeting, Washington, DC, August 2017.

Progress 10/20/15 to 09/30/16

Outputs
Target Audience:This project will target American consumers to help them to selectdaily diets to improve their health. We will also help breeders to produce foods with enhanced amounts of anti-obesity beneficial compounds. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?A student, Yen-Chen Tung working on the anti-obesity research project has finished her thesis and obtained her Ph.D. degree in 2015. Another student, Hui-Yun Tsai also working on obesity-induced cancer and its prevention has successfully obtained her Ph.D. degree in 2016. How have the results been disseminated to communities of interest?The research results have been presented at the Fall ACS meeting in Philadelphia in 2016, as well as the annual meeting of International Society of Nutraceutials and Functional Foods at Orlando, Florida in 2016. What do you plan to do during the next reporting period to accomplish the goals?We are and will continue working on the anti-obesity effect of phytochemicals such as garcinol from Garcinia fruit, tetrahydrocurcumin from tumeric and theasinensins from Oolong tea.

Impacts
What was accomplished under these goals? Energy imbalance is the major cause of obesity. Adipogenesis is that excess calories store as triacylglycerol (TG) and increases the number and size of adipocytes. In lipogenesis, the syntheses of fatty acid and TG proceed in the liver and adipose tissues and eventually expand the adipose tissues.The function of brown adipose tissues is different from that of white adipose tissues (WAT) in the body. WAT is not only a major TG storage tissue that can be deposited under the skin and in the intra-abdominal area, but also an endocrine organ regulating the whole-body energy homeostasis. When our body keeps in positive energy balance, the adipose tissue dysfunction may develop ectopic fat, especially intra-abdominal fat will deposit in several organs, such as, liver, pancreas, and heart, and finally affect their functions.Therefore, obesity plays a key role in metabolic syndromes and is highly related to diabetes, cardiovascular disease, fatty liver disease and cancer.Transcription factors play a crucial role in controlling glucose, amino acid and lipid metabolic homeostasis. Sterol regulatory element-binding proteins 1 (SREBP-1) is an important transcription factor related to cellular lipogenesis, lipid homeostasis and adipocyte differentiation.In addition, AMPK is a major regulator of cellular energy balance through switch on the catabolic pathway and switch off the anabolic pathway.AMPK can regulate several metabolic pathways such as glucose transport, gluconeogenesis, lipogenesis and lipolysis.AMPK can control lipid metabolism by phosphorylating acetyl-CoA carboxylase (ACC)1 and 2 or regulate long-term effects on phosphorylated SREBP-1 and the loss of expression of lipogenic enzymes. Polymethoxyflavones (PMFs) and hydroxylated polymethoxyflavones (HPMFs) are flavonoids found in citrus peels and they have several biological benefits such as anti- inflammatory, anti-cancer activities and regulating lipid metabolism.Nobiletin and HPMFs can suppress lipid accumulation in 3T3-L1 preadipocytes and high-fat diet-induced obesity animal models,but the anti-adipogenesis ability of 5-OH-Nob is still unknown. It has been reported that both PMFs and HPMFs are highly lipophilic that gives them poor aqueous solubility and low oral bioavailability.Prodrugs are derivatives of drug molecules. During metabolism, the active parent drugs derived from prodrugs will be released. The formulation of a prodrug can improve aqueous solubility, chemical stability, pre-systemic metabolism and brain penetration. Chemical modification is one of the methods to apply on prodrug formation. Recently, Wang et al. used chemical modification to obtain 5-acetyloxy-6,7,8,4′-tetramethoxyflavone (5-acetylated 5-demethyltangeretin), which exhibited better anti-breast cancer ability than tangeretin. In our current study, based on the prodrug concept, we prepared 5-OH-Nob and 5-acetyloxy-6,7,8,3'4'-pentamethoxyflavone (5-acetylated 5-demethylnobiletin, 5-Ac-Nob) derived from nobiletin by chemical modification and their anti-adipogenesis activity in the 3T3-L1 preadipocytes and a high-fat diet-induced obese animal model, as well as the related molecular mechanisms were investigated.

Publications

• Type: Journal Articles Status: Published Year Published: 2015 Citation: Lai, C.S.; Liao, S.N.; Tsai, M.L.; Kalyanam, N.; Majeed, M.; Majeed, A.; Ho, C.T.; Pan, M.H. Calebin A inhibits adipogenesis and hepatic steatosis in high-fat diet-induced obesity via activation of AMPK signaling. Molecular Nutrition & Food Research 2015, 59, 1883?1895.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Lai, C.S.; Chen, Y.Y.; Lee, P.S.; Kalyanam, N.; Ho, C.T.; Liou, W.S.; Yu, R.C.; Pan, M.H. Bisdemethoxycurcumin inhibits adipogenesis in 3T3-L1 preadipocytes and suppresses obesity in high-fat diet-fed C57BL/6 mice. Journal of Agricultural and Food Chemistry 2016, 64(4), 821?830.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Weerawatanakorn, M.; Hung, W.L.; Pan, M.H.; Li, S.; Li, D.; Wan, X.; Ho, C.T. Chemistry and health-beneficial effects of oolong tea and theasinensins. Food Science and Human Wellness 2015, 4(4), 133?146.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Lee, M.F.; Lai, C.S.; Cheng, A.C.; Hou, J.S.; Badmaev, V.; Ho, C.T.; Pan, M.H. Krill oil and xanthigen separately inhibit high fat diet induced obesity and hepatic triacylglycerol accumulation in mice. Journal of Functional Foods 2015, 19(Part B), 913?921.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Guo, J.; Tao, H.; Cao, Y.; Ho, C.T.; Jing, S.; Huang, Q. Prevention of obesity and type 2 diabetes with aged citrus peel (Chenpi) extract. Journal of Agricultural and Food Chemistry 2016, 64(10), 2053?2061.