Diet Research Data：Effects of High Fat Diet 32 Feeding on Male C57BL/6J Mice

 

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１.Objective

The purpose of this study was to investigate the effects of feeding a high-fat diet (High Fat Diet 32, or HFD32) to male C57BL/6J mice.

 

２.Materials and Methods

（1）Experimental Site

The experiment was conducted in the mouse breeding room (conventional) of the Central Research Institute of Nippon Compound Feed Co., Ltd. (currently Feed One Co., Ltd.).

（2）Diets

• CE-2: Crude fat content 4.6%, Fat kcal% 11.9%, soluble non-nitrogenous substance content 51.4%, NFE kcal% 59.3%
• High Fat Diet 32 (HFD32): Crude fat content 32.0%, Fat kcal% 56.7%, soluble non-nitrogenous substance content 29.4%, NFE kcal% 23.2%

（3）Animals

Male C57BL/6JJcl mice were used as experimental animals (20 mice per group, 40 mice in total).

（4）Housing Conditions

• Temperature and humidity: Temperature = 21-25°C, Humidity = 40-60%
• Lighting: 12-hour light/dark cycle (lights on 9:00 AM - 9:00 PM)
• Cages: Individually housed in polycarbonate cages with sterilized wood chip bedding
• Diet: Ad libitum
• Drinking water: Ad libitum

（5）Experimental Procedure

C57BL/6J mice were introduced at 4 weeks of age and acclimatized for 1 week (fed CE-2). After acclimatization, mice were randomly divided into two groups (CE-2 group and High Fat Diet 32 group) based on body weight and blood glucose levels to ensure no significant differences between groups. The experimental diets were fed from 5 weeks of age to 20 weeks of age (16 weeks feeding period). After the feeding period, various parameters were measured. Statistical analysis was performed using Student's t-test.

 

３.Results

The results are described below.

 

1.Body Weight

Body weight changes

Body weight was measured weekly. Data points and vertical bars in the figure represent the mean ± standard error. Statistical analysis was performed between groups at each time point, and an asterisk (*) indicates a significant difference (p < 0.05).

 

2.Food and Water Intake

Food intake

Food intake is shown as the average daily intake per mouse at each time point. Data points and vertical bars in the figure represent the mean ± standard error. Statistical analysis was performed between groups at each time point, and an asterisk (*) indicates a significant difference (p < 0.05).

Water intake

Water intake is shown as the average daily intake per mouse at each time point. Data points and vertical bars in the figure represent the mean ± standard error. Statistical analysis was performed between groups at each time point, and an asterisk (*) indicates a significant difference (p < 0.05).

 

3.Fasting Blood Glucose Levels

Changes in Fasting Blood Glucose Levels

Blood was collected from the tail vein every 3 weeks, 4 hours after lights on (1:00 PM). Whole blood glucose levels were measured using a whole blood glucose meter (Glucotest PROR, Sanwa Kagaku Kenkyusho Co., Ltd.). Vertical bars in the figure represent the mean ± standard error. Statistical analysis was performed between groups at each time point, and an asterisk (*) indicates a significant difference (p < 0.05).

 

4.Oral Glucose Tolerance Test

Oral glucose tolerance test results at each time point - 1

The oral glucose tolerance test was performed at 5, 15, and 20 weeks of age (2-3 days, 10 weeks, and 15 weeks after starting the experimental diet, respectively; shown in the upper left, upper right, and lower left panels of the figure). After an overnight fast (20 hours), mice were orally administered a glucose solution (2 g/kg body weight) using a gavage needle. Blood was collected from the tail vein before administration (0 min) and 30, 60, and 120 min after administration. Whole blood glucose levels were measured using a whole blood glucose meter (Glucotest PROR, Sanwa Kagaku Kenkyusho Co., Ltd.). Data points and vertical bars in the figure represent the mean ± standard error. Statistical analysis was performed between groups at each time point, and an asterisk (*) indicates a significant difference (p < 0.05).

Oral glucose tolerance test results at each time point - 2

【Blood glucose sum】

The blood glucose sum was calculated by summing the blood glucose levels at each time point during the oral glucose tolerance test. Vertical bars in the figure represent the mean ± standard error. Statistical analysis was performed between groups at each time point, and an asterisk (*) indicates a significant difference (p < 0.05). Note that the results at 5 weeks of age represent 3 days after starting the experimental diet.

【Blood glucose increment area】

The blood glucose increment area was calculated using the following formula

Blood glucose increment area (mg/dl·h) = {(a + 2b + 3c + 2d) × 1/4} - 2a
(a, b, c, and d represent blood glucose levels at 0, 30, 60, and 120 min, respectively)

Vertical bars in the figure represent the mean ± standard error. Statistical analysis was performed between groups at each time point, and an asterisk (*) indicates a significant difference (p < 0.05). Note that the results at 5 weeks of age represent 3 days after starting the experimental diet.

 

5.Insulin Tolerance Test

Changes in insulin tolerance test results at each time point

The insulin tolerance test was performed at 10 and 20 weeks of age (5 and 15 weeks after starting the experimental diet, respectively). An insulin solution (0.75 IU/kg body weight) was administered intraperitoneally, and blood was collected from the tail vein before administration (0 min) and 20, 40, 60, 80, and 120 min after administration. Whole blood glucose levels were measured using a whole blood glucose meter (GlucoTest PROR, Sanwa Kagaku Kenkyusho Co., Ltd.). Data points and vertical bars in the figure represent the mean ± standard error. Statistical analysis was performed between groups at each time point, and an asterisk (*) indicates a significant difference (p < 0.05).

 

6.Blood Analysis at Biopsy

Concentrations of blood lipid-related substances and other components at Biopsy (20 weeks of age)

 

7.Weights of Major Organs at Biopsy

The values in the table represent the mean ± standard error. Animals were sacrificed at 20 weeks of age (n=8 per group). After a 20-hour fast, the abdomen was opened under ether anesthesia, and the weights of each organ were measured. The upper table shows the absolute weight, and the lower table shows the relative weight per 100 g body weight. Statistical analysis was performed between groups for each item, and an asterisk (*) indicates a significant difference (p < 0.05).

Weights of major organs and adipose tissues at Biopsy (20 weeks of age)

Relative weights of major organs and adipose tissues at Biopsy (20 weeks of age)