Positive Correlation Between Serum Taurine and Adiponectin Levels in High-Fat Diet-Induced Obesity Rats - Advances in Experimental Medicine and Biology - page 102

Table 11.1 Composition of experimental diet (g/100 g diet)
Normal diet (g) High-fat diet (g)
Casein 20 20
Corn starch 15 10
Sucrose 50 40
Cellulose 5 5
Bean oil 5 5
Lard 0 15
Vitamin mixture a 1 1
Mineral mixture b 3.5 3.5
DL-Methione 0.3 0.3
Choline bitartrate 0.2 0.2
a AIN-76 vitamin mixture (g/kg); thiamin hydrochloride 600 mg, riboflavin 600 mg, pyridoxine
hydrochloride 700 mg, nicotinic acid 3 g, D-calcium pantothenate 1.6 g, folic acid 200 mg, D -biotin
20 mg, cyanocobalamin 1 mg, retinyl palmitate premix (250,000 IU/g) 1.6 g, DL-alpha-tocopherol
acetate (250 IU/g) 20 g, cholecalciferol (400,000 IU/g) 250 mg, menaquinone 5 mg, sucrose, finely
powdered 972.9 g
b AIN-76 mineral mixture (g/kg); calcium phosphate dibasic 500 g, sodium chloride 74 g, potas-
sium citrate monohydrate 220 g, potassium sulfate 52 g, magnesium oxide 24 g, manganous car-
bonate (43 ~ 48%Mn) 3.5 g, ferric citrate (16 ~ 17%Fe) 6 g, zinc carbonate (70% ZnO) 1.6 g, cupric
carbonate (53 ~ 55% Cu) 0.3 g, potassium iodate 0.01 g, sodium selenite 0.01 g, chromium potas-
sium sulfate 0.55 g, sucrose finely powdered 118 g
epididymal fat (E-fat), and retroperitoneal fat (R-fat) were measured. The serums
were immediately frozen in liquid nitrogen, and then stored at −70°C until
analysis.
Level of serum taurine was analyzed using HPLC system (McMahon et al .
1996 ). Serum samples were mixed with acetonitrile and the supernatant was
adjusted to pH 9 by borate buffer. These solutions mixed with fluorescamine were
analyzed on the HPLC (Agilent Technologies 1200 series HPLC) and Waters C 18
reverse-phase column (250 × 4.6 mm i.d) at 385 nm. The mobile phases tetrahydro-
furan-acetonitrile-phosphate buffer (pH 3.5) (4:25:71, v/v/v, solvent A) and tetra-
hydrofuran-acetonitrile-phosphate buffer (pH 3.5) (4:24:72, v/v/v, solvent B) were
used to gradient elution with flow rate at 1 ml/min.
Levels of serum TG and total TC were analyzed using automatic analyzer (BPC
Biosed srl., Italy). HDL-C was obtained from the whole serum with high-density
lipoprotein precipitation reagent (AM204-1, Asan Pharmaceutical, Korea) after
precipitation of low-density lipoprotein and very-low-density lipoprotein for 10 min
at 3,000 rpm (Hettich Mikro 200R, Tuttlingen, Germany) (Warnick and Albers 1978 )
and then analyzed for HDL-C using the same method as with TC. Standard serum
(Asan Pharmaceutical, Korea) was used for calibration before every parameter was
analyzed. All of the results were expressed as mg/dl serum.
Serum adiponectin and leptin levels were determined by enzyme-linked immu-
nosorbent assay (ELISA) kit and radio immune assay (RIA) kit in Samkwang
Medical laboratories (Seoul, Korea), respectively.