Open Journal of Genetics, 2013, 3, 9-86 OJGen http://dx.doi.org/10.4236/ojgen.2013.32A2002 Published Online July 2013 (http://www.scirp.org/journal/ojgen/) UP780, a Chromone-Enriched Aloe Composition, Enhances Adipose Insulin Receptor Signaling and Decreases Liver Lipid Biosynthesis Julie Tseng-Crank1*, Seon-Gil Do2, Brandon Corneliusen1, Carmen Hertel1, Jennifer Homan1, Mesfin Yimam1, Jifu Zhao1, Qi Jia1 1Unigen, Seattle, USA 2Unigen Korea, Songjung-Ri, Byeongchen-Myeon, Vheonan-Si, Chungnam, South Korea Email: *[email protected] Received 12 May 2013; revised 17 June 2013; accepted 17 July 2013 Copyright © 2013 Julie Tseng-Crank et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. ABSTRACT 1. INTRODUCTION Nutrigenomic studies were conducted to uncover the Diabetes and prediabetes have become epidemic globally. mechanism of action for the hypoglycemic and insulin In recent US surveys, 26.9% of the population aged 65 sensitizing effects of UP780. From high fat diet-induc- years and older had diabetes and 35% of those 20 years ed obesity mouse model for UP780, livers and white or older had prediabetes [1]. WHO of the United Nations adipose tissues (WAT) from groups of lean control, estimated in 2011 that 346 million people worldwide had high fat diet (HFD), and HFD treated with UP780 were diabetes, and that number was likely to double by 2030 collected for microarray study. Microarray genera- [2]. Complications of diabetes include heart disease, ted gene expression changes were applied to Ingenu- stroke, hypertension, blindness, kidney disease, neuro- ity Pathway Analysis for changes in canonical meta- pathy, amputation, and dental disease. Economic bur- bolic and signaling pathways. Microarray was vali- den of diabetes was high, in US, the 2007 direct and in- dated by quantitative reverse transcriptase-polyme- direct cost of diabetes amounted to $174 billion [1]. rase chain reaction (QPCR), Western blots, liver tri- The culprit for the epidemic of type II diabetes and glyceride, liver cholesterol, liver steatosis, and insulin prediabetes/metabolic syndrome is obesity. Obesity pre- ELISA. UP780 treatment decreased liver gene expres- disposes patients to a complex metabolic pathology, cha- sions for multiple enzymes involved in fatty acid bio- racterized by a cluster of closely related clinical fea- synthesis and triglyceride production. UP780 treat- tures of central obesity, hyperglycemia, insulin resistance, ment increased gene expressions globally for the insu- dyslipidaemia and hypertension [3]. Metabolic syndrome lin receptor signaling pathway in WAT. Both liver is associated with increased risk of type II diabetes and triglyceride and liver cholesterol levels were signifi- cardiovascular disease; cardiovascular diseases ultima- cantly reduced by UP780 over HFD. The reduction of tely are responsible for a large proportion of diabetic liver fat was confirmed by microscopic analysis of li- mortality [4]. The epidemics in type II diabetes calls for ver steatosis. Finally, UP780 significantly decreased interventions at the stage of metabolic syndrome, with fasting plasma insulin level over HFD. The mechani- diet, exercise, and medications/dietary supplements tar- sm of action for UP780 indicated a reduction of liver geting insulin resistance, to decrease the societal burden fat accumulation and an enhancement in adipose tis- from diabetes. sue insulin signaling pathway. This provided mecha- For thousands of years, herbal medicine was the only nistic explanation for the in vivo UP780 effects of en- treatment available to mankind. What is old is not nec- hanced insulin sensitiveity and decreased blood glu- essarily worthless in the modern world. Recently, tradi- cose in mouse diabetes and prediabetes models. tional herbal medicines with a history in diabetic treat- ment, now as nutritional supplements or as alternate me- Keywords: Nutrigenomics; Insulin Signaling Pathway; dicines, were rediscovered by the medical world for obe- Liver Fatty Acid Biosynthesis; Liver Steatosis; Aloe Vera sity, metabolic syndrome, and diabetes: Aloe vera leaf *Corresponding author. gel [5], berberine [6,7], bitter melon [8], ginsenoside Rb1 OPEN ACCESS 10 J. Tseng-Crank et al. / Open Journal of Genetics 3 (2013) 9-86 [9], genistein [10], and cinnamon [11], to name a few. In traction, microarray, Ingenuity Pathway Analysis, real- addition, resveratrol from red grape was recently disco- time RT-QPCR, Western blot, liver triglyceride and cho- vered to improve obesity and aging related disorders lesterol assays, liver steatosis, and plasma insulin ELISA [12]. are described in the supplemental materials. Aloe vera inner leaf gel and whole leaf extract had been reported to produce a hypoglycemic effect in dia- 3. RESULTS betic animal models of alloxan-induced diabetic mice [13-15] and type I/type II diabetic rats [16,17]. UP780, 3.1. Microarray/QPCR and UP780 Mechanism the Aloe vera inner leaf gel extract standardized with of Action aloesin, was found to decrease fasting blood glucose lev- HFD feeding of C57BL/6J mice is known to induce gene els and insulin resistance in mouse diet-induced obesity expression variations in organs that respond to high ca- (DIO) model and db diabetic model [18]. Significantly, loric intake such as liver, fat, and muscle [12]. It was of UP780 was the subject of a double blind, placebo con- great interest to observe the effects of UP780 treatment trolled clinical pilot study of prediabetic patients. After over HFD-induced changes in gene expressions. Micro- eight weeks of treatment, UP780 significantly reduced array gene expression variations were applied to the con- fasting blood glucose, insulin, HbA1c, fructosamine, and struction of a hypothesis for UP780 mechanism of action, urinary F2-isoprostanes compared to placebo [19]. based on the canonical metabolic and signaling pathway Although aloe was traditionally used for diabetes analysis, and taken into account of known gene functions treatment, and shown to reduce blood glucose, improve [24,25]. The large volume of pathway analysis results lipid profile, and provide anti-oxidants in diabetic animal were allocated to the supplemental materials. Only key models [13,15-17,20,21], there is a paucity of aloe me- genes involved in the interpretation of biological signi- chanism of action study. From the UP780 prediabetic ficance of UP780 were validated by QPCR (Table 1). An DIO mouse study [18], livers and white adipose tissues equivalent Table 2 was generated for WAT. Concordance (WAT) collected from the DIO animals were used in a between liver microarray and liver QPCR was good, 11 nutrigenomic study. By microarray/pathway analysis and out of 14, when the genes loosely categorized as Sig- QPCR gene expression, UP780 decreased fatty acid bio- naling were not counted (Table 1) Concordance between synthesis and triglyceride production in liver and in- WAT microarray and QPCR was excellent, 11 out of the creased insulin receptor signaling pathway in WAT. The 13 key genes tested (Table 2) microarray and QPCR gene expression data was valida- The liver key gene list in Table 1 emphasized the me- ted by Western blot, liver triglyceride/cholesterol, liver tabolic pathways. Liver responds to the nutritional state steatosis, and plasma insulin ELISA. of the body by modifying the enzyme/gene activities in- volved in metabolism, either by protein phosphorylation 2. MATERIALS AND METHODS or by gene expression variation. With liver, genes encod- 2.1. Composition of UP780 ing relevant members of metabolic pathways of fatty acid biosynthesis, fatty acid metabolism, glycollysis/glu- UP780 is a combination of Aloe vera inner leaf gel coneogenesis, and fat transport were all found to be stri- ® powder (QMatrix ) standardized to contain 2% - 4% alo- kingly up-regulated by HFD and down-regulated by ® esin. QMatrix is produced from Aloe vera gel car- UP780 (Table 1). The hypoglycemic effect of UP780 bon-filtered to remove anthraquinone, which has laxative suggested a decreased gluconeogenesis pathway. Many effect [22]. Aloesin, an aromatic chromone isolated from genes in glycolysis/gluconeogenesis pathway were up- Aloe ferox, was discovered by high throughput screening regulated by HFD and down-regulated by UP780 (Table of 2059 plant extracts to increase adiponectin secretion 1, Supplement Figure 5 and Supplement Table 1). from differentiated 3T3-L1 adipocytes [18]. In the pre- While this may suggest a decrease of gluconeogenesis by paration of QMatrix®, aloesin was removed by carbon ® UP780, close examination of the functions of the genes filtration. Aloesin was added to QMatrix with an expec- involved indicated contradictions that cannot be resolved tation of broadening the aloe action for glycemic control. by gene expression data alone. In particular, phosphoe- GW1929 is a specific non-thiazolidinedione, tyrosine- nolpyruvate carboxykinase (PEPCK) is a rate-limiting derived agonist of peroxisome proliferatoractivated rece- enzyme for gluconeogenesis. The cytosolic PEPCK1 was ptor gamma (PPARα, Tocris Bioscience). Thiazolidine- up-regulated by HFD/UP780, opposing gluconeogenesis dione class PPAR agonists are standard medicines for as a mechanism of hypoglycemic control. Effect of type II diabetes [23]. UP780 on carbohydrate metabolism in liver will need further studies. 2.2. Methods Strong evidence of UP780-modulated metabolic