Intermittent Fasting Partially Alleviates Dietary Margarine-Induced Morphometrical, Hematological, and Biochemical Changes in Female Mice, but Not in Males

Abstract

Margarine is a popular high-calorie component of the Western diet and was shown to be associated with the development of metabolic syndrome. Intermittent fasting (IF) is an effective approach to improve health and prevent metabolic disorders. This study aimed to investigate the effects of margarine consumption, both ad libitum and in combination with IF regimens, using young C57BL/6J mice of both sexes. Female mice fed margarine ad libitum as a supplement to the standard diet showed significant body mass gain, reduced food intake, lower blood paraoxonase activity, and higher lipid peroxide (LOOH) levels, along with higher activities of antioxidant enzymes in the liver. Margarine-fed males showed higher food intake and had lower blood triacylglycerol levels, higher LOOH levels in adipose tissue, and lower LOOH levels in the liver than their control counterparts. When a margarine-supplemented diet was provided to mice with an IF regimen, males gained body mass faster and experienced severe metabolic changes, including elevated fasting blood glucose levels, higher total leukocyte count, triacylglycerol accumulation, and reduced glycogen levels in the liver compared to their margarine ad libitum counterparts. Females treated with margarine + IF showed a partial improvement in metabolic status and a decrease in proinflammatory markers compared to the group receiving margarine ad libitum. Hence, responses to the diets were sex-specific. Females that consumed margarine ad libitum had higher metabolic sensitivity than males. Meanwhile, IF provided some protective effects in females but worsened metabolic outcomes in males when combined with a high-fat margarine diet.

Keywords: every-other-day fasting; food consumption; inflammation; liver; metabolic health; sex differences; western diet.

Figure 1

Experimental design. One-month-old mice were randomly divided into three groups: (i) the control group was fed ad libitum a standard food (control), (ii) margarine group was fed ad libitum standard food supplemented with margarine (MarAD), and (iii) the margarine + intermittent fasting (MarIF) group was also fed standard food supplemented with margarine, but with an alternating schedule of 24 h of feeding followed by 24 h of complete fasting.

Figure 2

Changes in body mass in mice fed a standard food ad libitum (control), margarine-supplemented food ad libitum (MarAD), and margarine-supplemented food with an IF regimen for 16 weeks (MarIF). In each group, mice were housed in two cages, with three mice in one cage (n = 6 mice per group). Males and females were kept in separate cages. Dynamics of absolute body mass in males (a) and females (b), and percentage body mass gain at the end of the experiment (c). ^∗Significantly different from the value in the corresponding control group with p ≤ 0.05. ^∗∗Significantly different from the values in the corresponding MarAD group with p ≤ 0.05. ^#Significantly different between sexes with p ≤ 0.05.

Figure 3

Food and energy consumption by mice fed a standard food ad libitum (control), margarine-supplemented food ad libitum (MarAD), and margarine-supplemented food with an IF regimen (MarIF). Dynamics of daily food consumption (a, b), average daily food consumption (basic food is marked in blue, margarine is marked in dark blue) (c, d), dynamics of daily energy intake (e, f), average daily number of calories consumed (calories from basic food is marked in blue, and calories form margarine is marked in dark blue) (g, h), and average daily water consumption (i). Statistical information is shown in Figure 2.

Figure 4

Total and differential leukocyte counts in mice fed a standard food ad libitum (control), margarine-supplemented food ad libitum (MarAD), and margarine-supplemented food with an IF regimen (MarIF). The number of total leukocytes (a) and relative counts of band neutrophils (b), segmented neutrophils (c), monocytes (d), and lymphocytes (e). Statistical information is shown in Figure 2.

Figure 5

Blood biochemical parameters in mice fed a standard food ad libitum (control), margarine-supplemented food ad libitum (MarAD), and margarine-supplemented food with an IF regimen (MarIF). Levels of free glucose (a), triacylglycerols, TAG (b), total cholesterol (c), paraoxonase activity (d), myeloperoxidase activity (e), and interleukin-1β, IL-1β level (f). Statistical information is shown in Figure 2.

Figure 6

IL-1β levels and lipid peroxide (LOOH) levels in adipose tissue of mice fed a standard food ad libitum (control), margarine-supplemented food ad libitum (MarAD), and margarine-supplemented food with an IF regimen (MarIF). Levels of interleukin-1β, IL-1β (a), and lipid peroxides, LOOH (b). Statistical information is shown in Figure 2.

Figure 7

Energy metabolism indicators in the liver of mice fed a standard diet (control), margarine-supplemented food ad libitum (MarAD), and margarine-supplemented diet with an IF regimen (MarIF). Levels of glycogen (a), triacylglycerols (TAG) (b), and activities of phosphofructokinase (PFK) (c), and pyruvate kinase (PK) (d). Statistical information is shown in Figure 2.

Figure 8

Markers of oxidative stress and antioxidant defense in the liver of mice fed a standard food ad libitum (control), margarine-supplemented food ad libitum (MarAD), and margarine-supplemented food with an IF regimen (MarIF). Lipid peroxide (LOOH) levels (a), activities of superoxide dismutase, SOD (b), catalase (c), glutathione-S-transferase, GST (d), glutathione peroxidase, GPx (e), and glucose-6-phosphate dehydrogenase, G6PDH (f). Statistical information is shown in Figure 2.