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Glucosamine enhances body weight gain and reduces insulin response in mice fed chow diet but mitigates obesity, insulin resistance and impaired glucose tolerance in mice high-fat diet.

Metabolism: clinical and experimental (2014-12-18)
Ji-Sun Hwang, Ji-Won Park, Moon-Suk Nam, Hyeongjin Cho, Inn-Oc Han
ABSTRACT

This study investigated the potential of glucosamine (GlcN) to affect body weight gain and insulin sensitivity in mice normal and at risk for developing diabetes. Male C57BL/6J mice were fed either chow diet (CD) or a high fat diet (HFD) and the half of mice from CD and HFD provided with a solution of 10% (w/v) GlcN. Total cholesterol and nonesterified free fatty acid levels were determined. Glucose tolerance test and insulin tolerance test were performed. HepG2 human hepatoma cells or differentiated 3T3-L1 adipocytes were stimulated with insulin under normal (5 mM) or high glucose (25 mM) conditions. Effect of GlcN on 2-deoxyglucose (2-DG) uptake was determined. JNK and Akt phosphorylation and nucleocytoplasmic protein O-GlcNAcylation were assayed by Western blotting. GlcN administration stimulated body weight gain (6.58±0.82 g vs. 11.1±0.42 g), increased white adipose tissue fat mass (percentage of bodyweight, 3.7±0.32 g vs. 5.61±0.34 g), and impaired the insulin response in livers of mice fed CD. However, GlcN treatment in mice fed HFD led to reduction of body weight gain (18.02±0.66 g vs. 16.22±0.96 g) and liver weight (2.27±0.1 vs. 1.85±0.12 g). Furthermore, obesity-induced insulin resistance and impaired Akt insulin signaling in the liver were alleviated by GlcN administration. GlcN inhibited the insulin response under low (5 mM) glucose conditions, whereas it restored the insulin response for Akt phosphorylation under high (25 mM) glucose conditions in HepG2 and 3T3-L1 cells. Uptake of 2-DG increased upon GlcN treatment under 5 mM glucose compared to control, whereas insulin-stimulated 2-DG uptake decreased under 5 mM and increased under 25 mM glucose in differentiated 3T3-L1 cells. Our results show that GlcN increased body weight gain and reduced the insulin response for glucose maintenance when fed to normal CD mice, whereas it alleviated body weight gain and insulin resistance in HFD mice. Therefore, the current data support the integrative function of the HBP reflecting the nutrient status of lipids or glucose and further implicate the importance of the pathway in insulin signaling for the regulation of metabolism.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Cholesterol, tested according to Ph. Eur.
Sigma-Aldrich
α-D-Glucose, anhydrous, 96%
Sigma-Aldrich
Glucose solution, BioUltra, for molecular biology, ~20% in H2O
Sigma-Aldrich
D-(+)-Glucosamine hydrochloride, ≥99%, crystalline
Sigma-Aldrich
D-(+)-Glucosamine hydrochloride, ≥99%, BioReagent, suitable for cell culture
Sigma-Aldrich
Cholesterol, Sigma Grade, ≥99%
Sigma-Aldrich
Cholesterol, powder, BioReagent, suitable for cell culture, ≥99%
Sigma-Aldrich
Cholesterol, from sheep wool, ≥92.5% (GC), powder
Sigma-Aldrich
D-(+)-Glucosamine hydrochloride, Vetec, reagent grade, ≥99%
SAFC
Cholesterol, from sheep wool, Controlled origin, meets USP/NF testing specifications
Sigma-Aldrich
Cholesterol, Vetec, reagent grade, ≥92.5%
Supelco
Cholesterol solution, certified reference material, 10 mg/mL in chloroform
Supelco
Cholesterol, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
D-(+)-Glucose solution, 45% in H2O, sterile-filtered, BioXtra, suitable for cell culture
Sigma-Aldrich
D-(+)-Glucose solution, 100 g/L in H2O, sterile-filtered, BioXtra, suitable for cell culture
Supelco
D-(+)-Glucose solution, 1 mg/mL in 0.1% benzoic acid, standard for enzymatic assay kits GAGO20, GAHK20, STA20, analytical standard
Supelco
Glucosamine hydrochloride, Pharmaceutical Secondary Standard; Certified Reference Material
USP
Glucosamine hydrochloride, United States Pharmacopeia (USP) Reference Standard
SAFC
Cholesterol, Plant-Derived, SyntheChol®, NF, Ph. Eur., JP
Glucosamine for system suitability, European Pharmacopoeia (EP) Reference Standard
Glucosamine hydrochloride, European Pharmacopoeia (EP) Reference Standard