High-Fat Diet in Perinatal Period Promotes Liver Steatosis and Low Desaturation Capacity of Polyunsaturated Fatty Acids in Dams: A Link with Anxiety-Like Behavior in Rats

Abstract

Background/objectives: This study investigates the effects of a high-fat diet (HFD) during pregnancy and lactation on maternal and offspring health, focusing on behavioral, metabolic, and fatty acid composition outcomes in a rat model.

Methods: Twelve female Sprague-Dawley rats were fed either a control diet, CD (n = 6), or HFD (n = 6) for 12 weeks, encompassing mating, gestation, and lactation periods (18 weeks). Anxiety-like behavior, maternal behavior, depression-like behavior, and social play were studied. Post mortem, the liver function, hepatic steatosis, and fatty acid composition (erythrocytes, liver, adipose tissue) were evaluated. In regard to desaturase enzymes (Δ-6D and Δ-5D), liver activity, protein mass, and gene expression (RT-PCR) were analyzed. Additionally, gene expression of PPAR-α, ACOX, CPT1-α, SREBP-1c, ACC, and FAS was assessed. Statistical analysis was performed using Student's t-test, mean ± SD (p < 0.05).

Results: The HFD significantly increased maternal weight and anxiety-like behavior while reducing social interactions exclusively in male offspring (p < 0.05). It also led to a significant decrease in the synthesis and content of n-3 PUFAs in the analyzed tissues, induced hepatic steatosis, and upregulated the expression of pro-lipogenic genes in the maternal liver.

Conclusions: These findings suggest that long-term HFD consumption alters tissue fatty acid composition, disrupts metabolic homeostasis, and contributes to behavioral changes, increasing anxiety-like behaviors in pregnant dams and reducing social interactions in male offspring. Overall, this study provides further insight into the detrimental effects of HFD consumption during the perinatal period.

Keywords: alpha-linolenic acid; anxiety; docosahexaenoic acid; fatty acid metabolism; high-fat diet; linoleic acid; n-3 polyunsaturated fatty acid; pregnancy.

Figure 1

Experimental design. PND = post-natal day, LD = lactation day. Created with BioRender.com.

Figure 2

Maternal body weight gains and food intake in g/day and kcal/day during pre-pregnancy (A–C), pregnancy (D–F), and the postnatal period (G–I) in dams fed with control diet (CD), high-fat diet (HFD). Data are expressed as mean ± DS. n = 12. CD, control diet (n = 6); HFD, high-fat diet (n = 6). Significant differences between groups are indicated with *. Differences were calculated by two-way ANOVA followed by Sidák (p < 0.05) between CD (n = 6) and HFD (n = 6).

Figure 3

Adipose tissue and liver weights in mothers. (A) Offspring weight on PND7. (B) Offspring weight on PND21. (C) Dam’s tissue weight. (D) Tissue weights of whole litter offspring at PND7 and 21. (E) Male tissue weight at PND7 and 21. (F) Female tissue weight at PND7 and 21. Data are expressed as mean ± SD. Significant differences between groups are indicated with *. Differences were calculated by ANOVA two-way followed by Sidák (p < 0.05) between CD (n = 6) and HFD (n = 6).

Figure 4

Morphological characteristics in the liver of dams and offspring. Representative liver hematoxylin and eosin staining. Rats subjected to CD and HFD. Representative liver sections from a total of 6 animals per experimental group (hematoxylin–eosin; original magnification 10×). (A–E) images of Dams and offspring with CD, (F–J) images of Dams and offspring with HFD, (K) Score of steatosis in Dams, (L) score of steatosis at 7 PND, (M) score of steatosis at 21 PND. Significant differences between groups are indicated with *. Differences were calculated by ANOVA two-way followed by Sidák (p < 0.05) between CD (n = 6) and HFD (n = 6).

Figure 5

(A) Relative mRNA expression of peroxisome proliferator-activated receptor alpha (PPAR-α), Acyl-CoA oxidase (ACOX), carnitine palmitoyl transferase 1α (CPT1-α), sterol response element binding protein (SREBP-1c), Acetyl-CoA carboxylase (ACC), FA synthase (FAS). FADS-2 encodes Δ-6 Desaturase (Δ-6D) and FADS-1 encodes Δ-5 Desaturase (Δ-5D) in dam liver. (B,C) Quantification of Δ-6 Desaturase (Δ-6D) and Δ-5 Desaturase (Δ-5D) in dam liver. (D,E) Evaluation of Δ-6 Desaturase (Δ-6D) and Δ-5 Desaturase (Δ-5D) activity in dam liver. Data are expressed as mean ± SD. Significant differences between groups are indicated with *. Differences were calculated by ANOVA two-way followed by Sidák (p < 0.05) between CD (n = 6) and HFD (n = 6).

Figure 6

Relative mRNA expression in offspring liver at 7 and 21 days. (A–C) Expression of PPAR-α: Peroxisome proliferator-activated receptor alpha. (D–F) Expression of ACOX: Acyl-CoA oxidase. (G–I) Expression of CPT1-α: carnitine palmitoyl transferase 1α. (J–L) Expression of SREBP-1c: Sterol response element binding protein. (M–O) Expression of ACC: Acetyl-CoA carboxylase. (P–R) Expression of FAS: FA synthase. Data are expressed as mean ± SD. Significant differences between groups are indicated with *. Differences were calculated by two-way ANOVA followed by Sidák (p < 0.05) between CD (n = 6) and HFD (n = 6).

Figure 7

Relative mRNA expression in offspring liver at 7 and 21 days. (A–C) Expression of FADS-1, encodes Δ-5 Desaturase (Δ-5D). (D–F) Expression of FADS-2, encodes Δ-6 Desaturase (Δ-6D). Data are expressed as mean ± SD. Significant differences between groups are indicated with *. Differences were calculated by two-way ANOVA followed by Sidák (p < 0.05) between CD (n = 6) and HFD (n = 6).

Figure 8

Dam behavioral results. (A) Combined heatmap generated by EthoVision^® Software XT 18 version (Noldus, Wageningen, The Netherlands) of Elevated Plus Maze (EPM) test in dams. OA: Open Arm. (B) Time spent in the open arms in EPM test. (C) Time spent in the closed arms in EPM test. (D) Number of entries to open arm in EPM test. (E) Anxiety index calculated according to Neto et al. [15]. (F) Number of entries to the light box in the Light Dark Box test. (G) Percentage of nesting material used by dams. (H) Total time spent on maternal behavior. (I) Latency until first contact with offspring in pup retrieval test. (J) Latency until first offspring retrieval in pup retrieval test. Values are expressed as media ± SD. Differences were calculated by Unpaired t-test with Welch’s correction (p < 0.05) between CD (n = 6) and HFD (n = 6). Offspring behavioral results: (K) Maximum speed in Open Field Test (OFT). (L) Number of entries to the center zone in Open Field Test (OFT). (M) Time spent in central zone in OFT. (N) Prosocial behavior in offspring: Time spent interacting with a novel rat. (O) Sucrose preference test. Data are expressed as mean ± SD. Significant differences between groups are indicated with *. Differences were calculated by two-way ANOVA followed by Sidák (p < 0.05) between male CD (n = 6), female CD (n = 6), male HFD (n = 6), and female HFD (n = 6).