Diet Research Data：Effects of High Fat Diet 32 Feeding on Female 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 female C57BL/6J mice.

 

２.Materials and Methods

（1）Experimental Site

The experiment was conducted in the animal breeding facility (conventional) of the Japan Veterinary and Animal Sciences University (currently Japan Veterinary and Life Science University).

（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

Female C57BL/6JJcl mice were used as experimental animals (10 mice per group, 20 mice in total).

（4）Housing Conditions

• Temperature and humidity: Temperature = 21-25°C, Humidity = 40-60%
• Lighting: 14-hour light/10-hour dark cycle (lights on 6:00 AM - 8:00 PM)
• Cages: Individually housed in polycarbonate cages with sterilized wood chip bedding
• Diet: Ad libitum
• Drinking water: Ad libitum tap water

（5）Experimental Procedure

Female 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 based on body weight, blood glucose levels, and blood glucose levels from an oral glucose tolerance test at 5 weeks of age to ensure no significant differences between groups. The experimental diets were fed from 5 weeks of age to 20 weeks of age. 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

Whole blood was collected and 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

The oral glucose tolerance test was performed at 15 and 20 weeks of age (before starting the experimental diet, 10 weeks, and 15 weeks). 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).

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 (0.5 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

Blood lipid analysis results

Blood was collected at Biopsy (20 weeks of age) from each group (n=10). After a 20-hour fast, animals were anesthetized with ether, and blood was collected from the posterior vena cava. Serum levels of total cholesterol (T-Cho), HDL-cholesterol (HDL-Cho), triglycerides (TG), and non-esterified fatty acids (NEFA) were analyzed. Vertical bars in the figure represent the mean ± standard error. Statistical analysis was performed between groups for each item, and an asterisk (*) indicates a significant difference (p < 0.05).

 

7.Weights of Major Organs at Biopsy

Major organ weights

Animals were sacrificed at 20 weeks of age (n=5 per group). After a 20-hour fast, animals were anesthetized with ether, and organs were removed and weighed. Vertical bars in the figure represent the mean ± standard error. Statistical analysis was performed between groups for each item, and an asterisk (*) indicates a significant difference (p < 0.05).

Adipose tissue weights

Animals were sacrificed at 20 weeks of age (n=5 per group). After a 20-hour fast, animals were anesthetized with ether, and organs were removed and weighed. Vertical bars in the figure represent the mean ± standard error. 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 tissue

The values in the table represent the mean ± standard error. Statistical analysis was performed between groups for each item, and an asterisk (*) indicates a significant difference (p < 0.05).

8.Photographs of Liver and Visceral Adipose Tissue

Photographs of liver and visceral adipose tissue (HE staining, ×400)

The upper panel shows the liver, and the lower panel shows the adipose tissue.