Recipient Organization
Jersey Flora, Inc
30 Outwater Ln
Garfield,NJ 07026
Performing Department
(N/A)
Non Technical Summary
More than 20% of the American population harbor the risk factors defined for metabolic syndrome, a condition that is highly correlated with the development of type-2 diabetes and coronary heart disease. Various botanical extracts have been used in traditional medicines to treat the symptoms of diabetes, and many of these preparations contain saponins. We have isolated an extract from the bulbs of Lilium longiflorum containing saponins and polyphenols that reduced blood glucose and body weight in laboratory mice on a high fat diet. We propose to test our more highly purified extracts and pure compounds in a transgenic type-2 diabetes mouse model (db/db) to determine the active ingredients in the lily bulb extracts. Since bulbs of Lilium sp. are edible, this research could lead to the development of L. longiflorum bulbs, or bulb extracts, as a functional food that may be consumed regularly to prevent the development of metabolic syndrome and the onset of diabetes.
Animal Health Component
60%
Research Effort Categories
Basic
20%
Applied
60%
Developmental
20%
Goals / Objectives
Long-term goal: To develop lily bulbs or extracts from the bulbs as a functional or medical food. Short-term objective (Phase 1): To determine if purified extracts containing the identified steroidal glycosides in L. longiflorum have a hypoglycemic and/or hypolipidemic effect in laboratory mice.
Project Methods
Freeze-dried L. longiflorum bulbs are washed with pentanes, centrifuged and the pellet extracted with ethanol:water (70:30). The extract is concentrated in vacuo and freeze-dried. The 70:30 extract is reconstituted with water and solvent partitioned with ethyl acetate. The aqueous fraction is saved and partitioned with butanol. The butanol fraction is saved, concentrated in vacuo and freeze-dried. The butanol fraction is purified by silica gel column chromatography with a step gradient of chloroform-methanol, followed by LH-20 size exclusion chromatography and semi-prep reverse-phase HPLC. Pure compounds of the 5 identified steroidal glycosides are obtained by precipitation of the collected fractions. ESI LC/MS using an acetonitrile-water gradient will be used to monitor steroidal glycoside purification. The hypoglycemic/hypolipidemic effect of lily bulb extracts will be determined with a db/db mouse model using female 4-week-old weanling C57BL/KsJ (BL/Ks) homozygous diabetic (db/db) and heterozygous lean (db/m or "ln") control mice (Jackson Laboratory, Bar Harbor, ME). The effects of lily bulb extract on blood glucose and insulin levels, total triglycerides, total cholesterol, abdominal fat and body weight will be evaluated after 6 weeks/experiment. Six mice (4-5 weeks old) will be placed in each plastic cage and will be fed Purina Laboratory Chow 5001 diet (Ralston-Purina, Co., St Louis, MO) ad libitum. The mice will be kept on a 12-h light/12-h dark cycle in the animal facility of the Laboratory for Cancer Research, School of Pharmacy, Rutgers University. In a typical experiment, mice will be divided into 3 groups, 6 mice per group: i) untreated control, + water; ii) mice receive a low dose of lily bulb extract in their water; iii) mice receive a high dose of lily bulb extract in their water. After an overnight fast, animals will be sacrificed. Measurements: Blood glucose levels in mice will be determined by glucose oxidase. Blood insulin levels will be determined by enzyme linked immunosorbent assay (ELISA) using a kit (Boehringer Mannheim Diagnostic, Mannheim, Germany). Serum will be analyzed for cholesterol, triglycerides and HDL-cholesterol using commercially available kits. Atherogenic Index will be calculated as: Total cholesterol - HDL-cholesterol/HDL-cholesterol. Statistics: All data are expressed as mean plus or minus the SE. The Student's t test and analysis of variance (ANOVA) will be used for statistical analysis. The values will be considered significant when p value is less than 0.05. The research described in this proposal will provide efficacy and safety data in a mouse model for the development of lily bulb nutritional supplements, functional foods and beverages that may reduce blood glucose and the symptoms of type 2 diabetes so prevalent in Americans today.