2.4. Tissue measurements
Lipids were extracted from tissue homogenized with an
Omni Mixer Homogenizer in a 2:1 (v/v) chloroform–methanol
solution, according to Folch et al., 1957. Total lipids in this
homogenate were determined by the sulfophosphovanillin
method (Frings and Dunn, 1970; Meyer and Walther, 1980).
This method consists of oxidizing cellular lipids to small fragments
after chemical digestion with hot concentrated sulfuric
acid. After the addition of a solution of vanillin and phosphoric
acid, a red complex is formed that is measured with a spectrophometer
at 530 nm. The levels of total cholesterol were
measured by the reactions of cholesterol esterase, cholesterol
oxidase, and peroxidase enzymes (Labtest Kit/Liquiform).
Results are expressed as mg/g of tissue.
Glycogen was extracted from tissues following the method
described by Van Handel (1965), and glycogen levels in the
animals were determined as glucose equivalent (glucose–oxidase
method), after acidic hydrolysis (HCl) and neutralization
(Na2CO3), following the method of Geary et al. (1981). Glucose
was quantified using a Biodiagnostic kit (glucose–oxidase).
Results are presented as mmol/g of tissue.
Proteins were measured as described by Lowry et al. (1951),
with bovine serum albumin (Sigma) as the reference standard.
The results were expressed in mg/mL of homogenate.
Cholesterol Liquiform), with the results expressed in mg/mL.
Triglycerides were measured using a Biodiagnostic Kit (GPO
Trinder), and are expressed in mg/L.
2.4. Tissue measurementsLipids were extracted from tissue homogenized with anOmni Mixer Homogenizer in a 2:1 (v/v) chloroform–methanolsolution, according to Folch et al., 1957. Total lipids in thishomogenate were determined by the sulfophosphovanillinmethod (Frings and Dunn, 1970; Meyer and Walther, 1980).This method consists of oxidizing cellular lipids to small fragmentsafter chemical digestion with hot concentrated sulfuricacid. After the addition of a solution of vanillin and phosphoricacid, a red complex is formed that is measured with a spectrophometerat 530 nm. The levels of total cholesterol weremeasured by the reactions of cholesterol esterase, cholesteroloxidase, and peroxidase enzymes (Labtest Kit/Liquiform).Results are expressed as mg/g of tissue.Glycogen was extracted from tissues following the methoddescribed by Van Handel (1965), and glycogen levels in theanimals were determined as glucose equivalent (glucose–oxidasemethod), after acidic hydrolysis (HCl) and neutralization(Na2CO3), following the method of Geary et al. (1981). Glucosewas quantified using a Biodiagnostic kit (glucose–oxidase).Results are presented as mmol/g of tissue.Proteins were measured as described by Lowry et al. (1951),with bovine serum albumin (Sigma) as the reference standard.The results were expressed in mg/mL of homogenate.Cholesterol Liquiform), with the results expressed in mg/mL.Triglycerides were measured using a Biodiagnostic Kit (GPOTrinder), and are expressed in mg/L.
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