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Cell culture and AMPK assay by Western blot analysisHPLC analysis of active compounds and myristicin under different extraction conditionsExtraction and isolation of active compounds on AMPK from Myristica fragransProcessing procedures for reducing myristicin contents from extracts of Myristica fragransAnimal experiments
Le Thi Van Thu Advisor: Prof. Oh Won-Keun Ph.D Department of Pharmacy, Graduate School of Chosun University October, 2009 Thesis for Master Degree Metabolic syndrome WHO definition of the metabolic syndrome, 1999 Glucose intolerance or diabetes mellitus and/or insulin resistance together with two or more of the other components listed below: • Impaired glucose regulation or diabetes; • Insulin resistance • Raised arterial pressure ≥ 140/90 mmHg; • Raised plasma triglycerides and/or low HDL-cholesterol • Central obesity • Microalbuminuria Consequences of the syndrome Cardiovascular disease, hypertension and chronic renal disease Diabetes and obesity 1.1 billion adults worldwide are overweight 312 million are obese Modern diet Sedentary lifestyle resulted in an increase in obesity, type 2 diabetes,metabolic disorders “exercise in a pill” – AMPK activators AMP activated protein kinase: a target for total metabolic control Structure and regulation of AMPK Acyl CoA carboxylase and fat metabolism Myristica fragrans Family : Myristicaceae Common name: nutmeg Constituents: lignans, essential oil Biological activity: anti-oxidant, anti-inflammatory, anticariogenic, hepatoprotective activities. PTP1B inhibitory effect. Uses: spice, cosmetic, drugs. Part used: seed Cell culture and AMPK assay by Western blot analysis HPLC analysis of active compounds and myristicin under different extraction conditions Extraction and isolation of active compounds on AMPK from Myristica fragrans Processing procedures for reducing myristicin contents from extracts of Myristica fragrans Animal experiments M. Fragrans (seeds, 3 kg) Ethanol extract Extracted with 30% EtOH (30 L × 2, 1 week) OP-CC (10 × 60 cm) , HP-20 Mob. Phase: EtOH:H 2 O 60% EtOH 80% EtOH 90% EtOH 100% EtOH Aceton 60:40 80:20 90:10 100:0 Acetone F1 F2 F3 F5 F4 OP-CC (6 × 60 cm), Silica gel (63 - 200 µm) Sol. System: n-Hexan: Aceton (6:1 - 0:1) Compound 6 (12 mg) Compound 7 (8.5 mg) Compound 1 (370 mg) Compound 2 (28.9 mg) Compound 3 (12.7 mg) Compound 4 (6.7 mg) Compound 5 (7.8 mg) OP-CC (5 × 60 cm) RP ODS-A (12 nm) MeOH: H 2 O 1.5:1 to 2:1 OP-CC (5 × 60 cm) RP ODS-A (12 nm) MeOH: H 2 O 2:1 to 5:1 Gilson HPLC RP-C18 (10 × 250 mm, 10 µm) Mob. Phase: MeOH/H 2 O + 0.1% Formic acid Isocratic: 60/40 Structure determination of tetrahydrofuran type-lignans from Myristica fragrans 1 H-NMR 13 C-NMR Compound 1 Structure determination of tetrahydrofuran type-lignans from Myristica fragrans 1 H-NMR 13 C-NMR Compound 2