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Figure 1. Exercise and blood sampling plan for 7 human male subjects pre-administered with placebo, sesamin or vitamin E.

Blood sampling Before

Figure 1. Exercise and blood sampling plan for 7 human male subjects pre-administered with placebo, sesamin or vitamin E.

Animal study At rest

24 ddY mice were randomly divided into sesamin, vitamin E or control groups. lOOmg/kg of sesamin (S100), and lOOmg/kg of vitamin E (El00) was administered to the respective group via an orogastric tube while the control received the vehicle (1% carboxymethyl cellulose sodium salt [CMC]). Two hours following feeding, the animals were sacrificed for blood collection and liver excision.

The livers were washed in cold saline, homogenized in 40mM Tris-HCL (pH 7.4) containing 145 mM KC1, 4 mM EDTA and 5 mM dithiothreitol. Samples were then centrifuged at 8500g for 10 minutes to remove cell debris and nuclei and the supernatant retained for enzyme activity assay and protein determination.

Heparinized blood was obtained from the heart, and centrifuged to obtain plasma, which was then stored at -80°C for lipid peroxide (LPO) analysis [14].

During exercise

32 ddY mice were subjected to swimming exercise in an aquarium tank (35°C) 2 hrs after orogastric feeding of 10 mg/kg sesamin (S10), 100 mg/kg sesamin (S100), 100 mg/kg vitamin E (El00) dissolved in 1 % CMC. Control group received only CMC solution. Following 30 mins of continuous swimming, the animals were sacrificed by cervical dislocation and heparinized blood samples collected from the heart.

Enzyme assay

Total GPX activity was determined by a modified Paglia and Valentine [15] coupled assay procedure, using tert-butyl hydroperoxide substrate. Se-GPX activity was measured with the same procedure, using an H202 substrate. GST activity against Chloro-2,4-dinitrobenzene (CDNB) was also determined as described by Habig and Jakoby [16]. BCA assay was used to determine protein content of the enzyme source [17].

Measurement of LPO

Lipid hydroperoxide values were measured using the Hb-MB method, which detects hydroperoxide form of lipid peroxidation [18, 19].

Statistical analysis

Statistical analysis was perforated using the SPSS version 10.0 Package. Two-way analysis of variance was performed followed by Tukey's multiple comparison test. Data is presented as mean ± SEM.

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