Objectives: This study investigated the short- and long-term metabolic effects of two β-sitosterol with propionic acid cocrystals (CCA and CCB) compared with commercial β-sitosterol in male hamsters. Methods: Five experiments were conducted, including three acute and two chronic dietary interventions. Acute experiments evaluated the metabolic response to a single dose of commercial β-sitosterol (S) or its cocrystals (513 mg/kg) administered with a pork-fat load following 40% caloric restriction. Blood samples were collected before and 5 h after the lipid challenge. Long-term studies assessed daily supplementation with CCA (264 mg/kg) or CCB (at two doses, 114 and 342 mg/kg) during exposure to high-fat (HF) or Western diets (WD), respectively, for 21 days. Results: Acutely, CCA attenuated postprandial triglyceride increases in adult animals, while CCB induced a reduced rise in postprandial cholesterol across the three studies compared to S. Young animals treated with CCA and CCB exhibited a reduction in postprandial glucose levels. Long-term, in HF-fed hamsters, CCA lowered plasma cholesterol and triglyceride levels on day 14 and, at the endpoint, decreased liver weight and increased circulating sitosterol levels, unlike its commercial counterpart. In WD-fed animals, CCB supplementation dose-dependently elevated plasma sitosterol levels. At the low dose, CCB treatment reduced body weight gain and cholesterol increase and improved adipocyte morphology, whereas the high dose reduced diet-induced hepatic detrimental outcomes, showing a stronger hepatoprotective effect compared to S-treated animals. Together, these results indicate improved postprandial lipid handling in the short-term and a modulation of metabolic adaptation to obesogenic diets in the long-term. Conclusions: β-sitosterol with propionic acid cocrystals improved metabolic responses more effectively than commercial β-sitosterol, enhancing bioavailability and potential therapeutic value against diet-induced metabolic disturbances.

Maternal consumption of a Western diet (WD) during critical developmental periods has been shown to program long-term metabolic dysfunction in offspring. However, it remains unclear whether these early exposures leave a persistent transcriptomic imprint into adulthood and whether this signature can be reshaped by postnatal interventions, limiting insight into underlying mechanisms and opportunities for early intervention. Here, we aimed to identify long-term transcriptomic signatures associated with maternal WD exposure during gestation and lactation using peripheral blood mononuclear cells (PBMCs) from adult male and female offspring. We further evaluated the potential of two early-life interventions - maternal dietary normalization and pup leptin supplementation during lactation - to reverse these alterations. The offspring from dams fed a normal-fat diet during gestation and lactation was used as a control group. Considering all groups, the expression of 90 genes differed between the control and WD offspring. Both interventions normalized the expression of most genes altered by maternal WD exposure (83 and 76 out of 90, respectively), shifting the global transcriptomic profile toward control levels. Multivariate analyses identified the 20 highest-ranked contributors to the PLS-DA model distinguishing offspring of WD-fed dams from controls, with stronger discriminatory capacity in males than in females, suggesting coordinated dysregulation of immunometabolic pathways, primarily involving immune and inflammatory signaling, cell fate regulation, and metabolic homeostasis. In conclusion, interventions during lactation may effectively reverse most transcriptomic alterations associated with adverse maternal conditions, supporting this period as a critical window for postnatal reprogramming. These findings suggest that early nutritional interventions may help mitigate long-term health risks associated with adverse maternal diet and support the use of PBMC transcriptomic signatures as potential minimally invasive biomarkers of developmental programming.

Dietary bioactive compounds can modulate mammary gland gene expression, potentially influencing milk composition and infant metabolic programming. This study investigated whether selected dietary bioactive compounds modulate leptin and adiponectin expression in rat mammary epithelial organoids (MEO), a 3D in vitro model of the mammary gland. 18 nutrients were initially screened for effects on leptin gene expression, followed by confirmatory and dose-dependent analyses focusing on oleic acid (OA), leucine (Leu), betaine (Bet), and their triple combination. mRNA levels of leptin, adiponectin, and β-casein (Csn2), as well as selected genes related to one-carbon and lipid metabolism and specific microRNAs, were analyzed. OA, alone or in combination, consistently increased leptin, adiponectin, and Csn2 mRNA expression in MEO compared with controls. These effects were accompanied by downregulation of lipogenic markers, including Srebp1, and increased Cpt1b expression under the triple-combination treatment, without significant changes in miRNA levels. In contrast, Bet and Leu showed more variable, context-dependent effects on leptin, adiponectin, and Csn2 expression, while altering miRNA profiles that differed from those observed under OA-containing treatments. Overall, the results indicate that dietary bioactive compounds modulate gene expression of rat MEO.

Perinatal nutrition influences offspring metabolic phenotype and obesity risk. This study examines how a maternal obesogenic diet and offspring's leptin supplementation during suckling affect the metabolome in offspring adulthood, offering insights into long-term metabolic programming. Offspring of dams fed a standard diet (SD), a Western-style diet (WD) before and during gestation and lactation (WD-dams), or a WD switched to SD during lactation were supplemented throughout suckling with leptin or vehicle and then maintained on an SD post-weaning. Metabolomic analysis of plasma from offspring adult animals revealed sex-specific alterations in the relative abundance of circulating metabolites due to maternal WD. Among these, lower abundance of orotic acid in males and arginine in females emerged as the most discriminating metabolites between experimental groups influenced by maternal WD during gestation and lactation. These changes, indicative of poor metabolic programming, were largely normalized by dietary intervention during lactation. Leptin supplementation during suckling also induced favorable metabolic adaptations, reflected in lower N-acetylneuraminic acid and choline abundance (both sexes), higher 5-amino valeric acid betaine (males), lower TCA cycle intermediates (fumarate, succinate, malate), and elevated γ-glutamyl peptides (γ-glutamylalanine, γ-glutamylglutamine) (females), suggesting potential anti-inflammatory effects. These findings enhance our understanding of how early-life conditions influence long-term metabolism and underscore the potential of circulating metabolites as biomarkers for assessing perinatal interventions.

ABSTRACT Maternal obesity during the perinatal period promotes adverse metabolic programming in offspring, increasing susceptibility to metabolic disorders later in life. Myo‐inositol (MI), a bioactive compound abundant in breast milk, has shown short‐term benefits when supplemented during lactation. This study evaluated the long‐term effects of maternal MI supplementation during lactation on metabolic and cognitive health in offspring born to diet‐induced obese or standard chow diet (SD)‐fed dams, all challenged with a Western diet (WD) in adulthood. Wistar rats were fed a SD or WD from 1 month before mating and through gestation and lactation and treated with MI (200 mg/kg/day) or vehicle during lactation. Offspring were fed a SD from weaning to 4 months of age, followed by a WD until sacrifice at 6 months. In male offspring, maternal MI supplementation improved insulin sensitivity (lower insulin levels and HOMA‐IR), increased circulating BDNF (WD group), and enhanced cognitive performance. It also increased adipose GLUT4 levels and Bdnf expression and, in the liver, downregulated lipogenic genes, upregulated Bdnf, and reduced lipid content, with sex‐specific differences. These effects were accompanied by lower circulating levels of triglycerides, nonesterified fatty acids, and TNF‐α. Collectively, these findings indicate that maternal MI supplementation during lactation enhances offspring metabolic resilience and cognitive outcomes through integrated improvements in insulin signaling, lipid metabolism, and inflammatory regulation, partially mitigating the detrimental effects of maternal obesity and adult obesogenic diet in a sex‐specific manner.

Interscapular brown adipose tissue is surrounded by a white fat depot, the interscapular WAT (isWAT), which has received little attention. Here, we have characterized isWAT and the impact of chronic intake of a very high-fat diet, the cafeteria diet, on isWAT browning capacity. The effect of high-fat diets on WAT browning is controversial, however, some groups have described increased WAT UCP1 expression, probably to counteract increased adiposity. We analyzed mRNA expression of key brown/brite markers and lipid metabolism genes, and of selected proteins in isWAT of control, cafeteria-obese and post-cafeteria rats. Morphological/immunohistochemical characterization was also performed. Results present isWAT as a depot with a molecular profile close to that of browning-prone white fat depots (inguinal and retroperitoneal), which also shares key features with BAT, including UCP1 expression (mRNA/protein). The cafeteria diet intake induced mRNA expression of browning/thermogenic genes, which was restored after the diet was withdrawn, and increased expression of key genes involved in lipid catabolism in isWAT. Moreover, basal mRNA expression of brown/brite markers was greater than in other white depots and higher UCP1 protein levels were observed compared to the browning-prone retroperitoneal depot. Thus, isWAT appears as an interesting depot to be taken into consideration for lipid research and to better characterize browning regulation.

Brain-derived neurotrophic factor (BDNF) and leptin are essential in neurodevelopment and central regulation of feeding and energy balance. We studied the metabolic imprinting effects of physiological leptin supplementation during suckling in the brain of 5-week-old mouse pups. Leptin-treated animals showed lower cumulative food intake and increased energy efficiency, which was related to higher lean mass. Among different brain areas, hypothalamic expression of Bdnf and upstream transcription control-related genes, such as Ppargc1a and Fndc5, was increased by leptin supplementation, especially in females. This was accompanied by higher expression of energy balance key genes (such as Prkaa2 and Cpt1c) and insulin/leptin signalling pathways, primarily in females, also with lower levels of total/phosphorylated AMPK, ACC or STAT3, mainly in males. In leptin-treated females, the exon IV Bdnf promoter showed increased methylation at a specific CpG site. Leptin supplementation during suckling can sex-dependently imprint hypothalamic gene expression, regulating the Ppargc1a/Fndc5/Bdnf pathway and related genes involved in energy balance, associated with a leaner phenotype, with a higher positive impact in females.

ABSTRACT Breast milk provides a wide range of nutritional and bioactive components crucial for infant growth and development during lactation, ultimately influencing future health outcomes. Here, we used untargeted metabolomics to identify differences in the breast milk metabolome related to maternal overweight/obesity and diet, assessing their influence on infant weight gain in the first month of lactation. Anthropometric data from 52 lactating mother–infant pairs were collected at birth and 1 month postpartum, along with breast milk samples. Information about food consumed the day before milk sampling (24‐h questionnaire) was collected, and a food frequency questionnaire was completed. Our findings reveal that mothers with overweight/obesity had greater lactose levels (p = 0.025) and lower levels of orotic acid (p = 0.002), 3‐indoxyl sulfate (p = 0.027), heneicosanoic acid (p = 0.040), and N1‐methylguanosine (p = 0.046) in milk than normal‐weight mothers. Notably, reduced orotic acid levels were associated with a greater infant weight gain during this period. Furthermore, mothers with a low adherence to the Mediterranean diet showed lower levels of citric acid (p = 0.041), N6‐succinyladenosine (p = 0.043), uric acid (p = 0.034), and eicosenoic acid (p = 0.035), and higher levels of acylcarnitine C6:0 (p = 0.041) than mothers with a medium/high adherence. Besides orotic acid, other milk metabolites were related to infant weight gain during this period, with maternal fruit and fish consumption (p = 0.015 and p = 0.017) emerging as the most influential dietary factor for an adequate early infant growth trajectory. None of the metabolites identified remained statistically significant after multiple‐comparisons adjustment. Present results, showing how maternal diet and obesity can affect breast milk metabolites and infant growth, offer insights to improve lactation‐feeding practices and support healthy infant development.

Myo-inositol (MI) supplementation has emerged as a promising intervention to mitigate the malprogramming effects associated with adverse maternal conditions during the perinatal period. This study aimed to assess the effects of MI supplementation during lactation on metabolism in diet-induced obese rats and on early health outcomes in their offspring. Female Wistar rats were fed either a control (CON) or Western diet (WD) for one month before mating and during gestation and lactation. After parturition, dams were supplemented daily with either MI or vehicle (V) throughout lactation. Phenotypic traits, along with milk and blood parameters were analyzed in dams and their offspring during lactation. At weaning, hepatic lipid content and gene expression in the liver, retroperitoneal white adipose tissue (rWAT), and mammary gland were assessed in dams. MI supplementation significantly reduced hepatic lipid accumulation in WD-dams and decreased pro-inflammatory markers in the liver (Tnfa) and retroperitoneal white adipose tissue (rWAT) (Tnfa, Il6) in both CON- and WD-dams, as well as modulated milk insulin levels. Their offspring exhibited, in plasma, higher MI and BDNF concentrations as well as lower insulin levels, which are key factors in perinatal programming. These findings suggest that maternal MI supplementation during lactation confers metabolic benefits to both dams and their offspring, specifically improving maternal inflammation and lipid metabolism while influencing milk composition.

Early-life metabolic environment significantly impacts long-term cognitive and metabolic health. This study investigates transcriptomic alterations in the hippocampus and peripheral blood mononuclear cells (PBMC) of young rats exposed to an isocaloric high-fat diet (HFD), resulting in the metabolically obese, normal-weight (MONW) phenotype. Rats were pair-fed either a standard (NW group) or HFD (MONW group) for 11 weeks after weaning. Another group (MONW-Lep) received leptin supplementation during lactation and subsequently HFD. Transcriptomic analysis of the hippocampus showed disruption of pathways linked to obesity and cognitive decline in the MONW group, which were attenuated by leptin intake. This was consistent with the results of working memory (T-maze test), impaired in MONW versus NW, but preserved in MONW-Lep animals. PBMC transcriptomics revealed overlapping genes with the hippocampus. Notably, Piwil1, a gene linked to neurodegeneration, metabolic syndrome and obesity, was up-regulated in PBMC of MONW but not of MONW-Lep animals, reflecting early hippocampal changes and leptin's preventive effect. These findings highlight the influence of early nutrition on cognitive health, the protective potential of leptin counteracting the effects of HFD intake and the usefulness of PBMC as a reliable source of biomarkers of brain health.

OBJECTIVE:We investigated how a maternal Western diet (WD) affects milk microRNA (miRNA) profile and associates with metabolic programming in adipose tissues in pups. We also explored the impact of betaine supplementation during suckling, as betaine levels are reported to be reduced in WD-fed dams' milk. METHODS:A microarray analysis was performed to profile miRNA expression in dams' milk. Betaine levels were measured in the milk of dams and the plasma of their offspring. We also analyzed the expression of miRNA target genes in white and brown adipose tissues through gene expression analysis. RESULTS:Our findings confirm decreased betaine levels in the milk of WD-fed dams and the plasma of their offspring. The miRNA screening identified 37 deregulated miRNAs (36 downregulated), with the following 6 as the most relevant: miR-223-3p; miR-32-5p; let-7i-5p; miR-140-5p; miR-29a-3p; and miR-29c-3p (downregulated). Some of their target genes were upregulated in brown and white adipose tissues, particularly those related to thermogenesis and browning. Betaine supplementation in pups demonstrated a slight protective effect in females by enhancing thermogenic capacity. CONCLUSIONS:Our results underscore the profound impact of a maternal WD on milk miRNA composition, potentially influencing gene expression, thermogenesis, and adiposity in the offspring, with sex-related differences.

This study examines the protective effects of myo-inositol supplementation during suckling on long-term negative outcomes caused by fetal energy restriction, using a metabolomics approach.Offspring of rats from both control and 25% gestational calorie-restricted dams received either myo-inositol or the vehicle during suckling and were exposed to a Western diet (WD) between 5 and 7 months of age. Metabolomics analysis of plasma samples at 7 months allowed the identification of 164 metabolites, revealing marked sex differences to gestational restriction. In males, maternal calorie restriction resulted in alterations in 19 metabolites, while only six metabolites showed significant variations in females, consistent with a lower impact of gestational calorie restriction (GCR) on the adult phenotype. Supplementation with myo-inositol normalized the levels of 16 metabolites in males and all six in females. Functionally, myo-inositol mostly targeted liver-associated functions in males and metabolic control functions in females. Mild/moderate GCR leads to significant changes in the metabolomic profiles of adult offspring, with males experiencing higher metabolic alterations. Early postnatal myo-inositol supplementation may be a promising strategy to alleviate the negative metabolic effect of maternal undernutrition. Sex-specific differences in the metabolomic response emphasize the necessity of considering both sexes for effective interventions.

Perinatal nutrition exerts a profound influence on adult metabolic health. This study aimed to investigate whether increased maternal vitamin A (VA) supply can lead to beneficial metabolic phenotypes in the offspring. The researchers utilized mice deficient in the intestine-specific homeobox (ISX) transcription factor, which exhibits increased intestinal VA retinoid production from dietary β-carotene (BC). ISX-deficient dams were fed a VA-sufficient or a BC-enriched diet during the last week of gestation and the whole lactation period. Total retinol levels in milk and weanling livers were 2- to 2.5-fold higher in the offspring of BC-fed dams (BC offspring), indicating increased VA supplies during late gestation and lactation. The corresponding VA-sufficient and BC offspring (males and females) were compared at weaning and adulthood after being fed either a standard or high-fat diet (HFD) with regular VA content for 13 weeks from weaning. HFD-induced increases in adiposity metrics, such as fat depot mass and adipocyte diameter, were more pronounced in males than females and were attenuated or suppressed in the BC offspring. Notably, the BC offspring were protected from HFD-induced increases in circulating triacylglycerol levels and hepatic steatosis. These protective effects were associated with reduced food efficiency, enhanced capacity for thermogenesis and mitochondrial oxidative metabolism in adipose tissues, and increased adipocyte hyperplasia rather than hypertrophy in the BC offspring. In conclusion, maternal VA nutrition influenced by genetics may confer metabolic benefits to the offspring, with mild increases in late gestation and lactation protecting against obesity and metabolic dysregulation in adulthood.NEW & NOTEWORTHY A genetic mouse model, deficient in intestine-specific homeobox (ISX) transcription factor, is used to show that a mildly increased maternal vitamin A supply from β-carotene feeding during late gestation and lactation programs energy and lipid metabolism in tissues and protects the offspring from diet-induced hypertrophic obesity and hepatic steatosis. This knowledge may have implications for human populations where polymorphisms in ISX and ISX target genes involved in vitamin A homeostasis are prevalent.

Metabolically obese, normal-weight (MONW) phenotype is characterized by visceral adiposity and obesity-related complications despite the absence of excess body weight. Early identification of this phenotype is crucial to establish preventive strategies. We aim to validate the utility of peripheral blood mononuclear cells (PBMC) transcriptome to detect metabolic risk related to the MONW phenotype at early life stages (young adulthood). Male Wistar rats were pair-fed either standard (NW group) or a high-fat diet (MONW group) after weaning, until 3.5 months. Global gene expression was examined by microarray in PBMC, and specific genes of interest by RT-qPCR in PBMC and liver. Results were validated in adult 6-month-old MONW rats. Young MONW animals had similar weight to controls (NW group) but greater adiposity, including liver fat content, and insulin resistance signs. PBMC transcriptome distinguished clearly MONW from NW rats. Neurological pathways were affected in line with impaired cognition in these animals. Most top-regulated genes were related to inflammation, including the top-up and down-regulated genes, Hpgds and Slfn4. Expression of fatty liver-related genes like Mkrn1 and Nampt was also affected in PBMC of the young MONW group mirroring liver alteration. Slfn4 and Mkrn1 appeared as especially relevant biomarkers with altered expression also in PBMC of adult 6-month-old MONW rats. In conclusion, PBMC transcriptomic analysis emerges as a tool for identifying early biomarkers of obesity-related metabolic risk in young and apparently healthy (lean) subjects, pointing towards increased inflammation, liver fat deposition, and cognitive alterations.

Placental leptin may impact foetal development. Maternal overnutrition has been linked to increased plasma leptin levels and adverse effects on offspring, whereas choline, an essential nutrient for foetal development, has shown promise in mitigating some negative impacts of maternal obesity. Here, we investigate whether a maternal obesogenic diet alters foetal growth and leptin levels in the foetal stomach, amniotic fluid (AF), and placenta in late gestation and explore the potential modulating effects of maternal choline supplementation. Female rats were fed a control (CD) or a western diet (WD) four weeks before mating and during gestation, half of them supplemented with choline (pregnancy days 11-17). Leptin levels (in foetal stomach, AF, and placenta) and leptin gene expression (in placenta) were assessed on gestation days 20 and 21. At day 20, maternal WD feeding resulted in greater leptin levels in foetal stomach, placenta, and AF. The increased AF leptin levels were associated with a premature increase in foetal weight in both sexes. Maternal choline supplementation partially prevented these alterations, but effects differed in CD dams, causing increased AF leptin levels and greater weight in male foetuses at day 20. Maternal choline supplementation effectively mitigates premature foetal overgrowth induced by an obesogenic diet, potentially linked to increased AF leptin levels. Further research is needed to explore the sex-specific effects.

The intake of high-fat diets (HFDs) and obesity are linked to cognitive impairment. Here, we aimed to investigate whether an early metabolically obese, normal-weight (MONW) phenotype, induced with an HFD in young rats, also leads to cognitive dysfunction and to evaluate the potential cognitive benefits of neonatal intake of leptin. To achieve this, Wistar rats orally received physiological doses of leptin or its vehicle during lactation, followed by 11 weeks of pair-feeding with an HFD or control diet post-weaning. Working memory was assessed using a T-maze, and gene expression in the hippocampus and peripheral blood mononuclear cells (PBMCs) was assessed with real-time RT-qPCR to identify cognition biomarkers. Young MONW-like rats showed hippocampal gene expression changes and decreased working memory. Animals receiving leptin during lactation presented similar gene expression changes but preserved working memory despite HFD intake, partly due to improved insulin sensitivity. Notably, PBMC Syn1 expression appears as an accessible biomarker of cognitive health, reflecting both the detrimental effect of HFD intake at early ages despite the absence of obesity and the positive effects of neonatal leptin treatment on cognition. Thus, the MONW phenotype developed at a young age is linked to cognitive dysfunction, which is reflected at the transcriptomic level in PBMCs. Neonatal leptin intake can partly counteract this impaired cognition resulting from early HFD consumption.

The early stages of life, especially the period from conception to two years, are crucial for shaping metabolic health and the risk of obesity in adulthood. Adipose tissue (AT) plays a crucial role in regulating energy homeostasis and metabolism, and brown AT (BAT) and the browning of white AT (WAT) are promising targets for combating weight gain. Nutritional factors during prenatal and early postnatal stages can influence the development of AT, affecting the likelihood of obesity later on. This narrative review focuses on the nutritional programming of AT features. Research conducted across various animal models with diverse interventions has provided insights into the effects of specific compounds on AT development and function, influencing the development of crucial structures and neuroendocrine circuits responsible for energy balance. The hormone leptin has been identified as an essential nutrient during lactation for healthy metabolic programming against obesity development in adults. Studies have also highlighted that maternal supplementation with polyunsaturated fatty acids (PUFAs), vitamin A, nicotinamide riboside, and polyphenols during pregnancy and lactation, as well as offspring supplementation with myo-inositol, vitamin A, nicotinamide riboside, and resveratrol during the suckling period, can impact AT features and long-term health outcomes and help understand predisposition to obesity later in life.

(1) Background: Cold exposure induces metabolic adaptations that can promote health benefits, including increased energy disposal due to lipid mobilization in adipose tissue (AT). This study aims to identify easily measurable biomarkers mirroring the effect of cold exposure on AT. (2) Methods: Transcriptomic analysis was performed in peripheral blood mononuclear cells (PBMCs) and distinct AT depots of two animal models (ferrets and rats) exposed to cold, and in PBMCs of cold-exposed humans. (3) Results: One week of cold exposure (at 4 °C) affected different metabolic pathways and gene expression in the AT of ferrets, an animal model with an AT more similar to humans than that of rodents. However, only one gene, Tlcd4, was affected in the same way (overexpressed) in aortic perivascular and inguinal AT depots and in PBMCs, making it a potential biomarker of interest. Subsequent targeted analysis in rats showed that 1 week at 4 °C also induced Tlcd4 expression in brown AT and PBMCs, while 1 h at 4 °C resulted in reduced Tlcd4 mRNA levels in retroperitoneal white AT. In humans, no clear effects were observed. Nevertheless, decreased PBMC TLCD4 expression was observed after acute cold exposure in women with normal weight, although this effect could be attributed to short-term fasting during the procedure. No effect was evident in women with overweight or in normal-weight men. (4) Conclusions: Our results obtained for different species point toward TLCD4 gene expression as a potential biomarker of cold exposure/fat mobilization that could tentatively be used to address the effectiveness of cold exposure-mimicking therapies.

This study investigates the impact of maternal nutrition during lactation on inflammation and oxidative stress in the offspring of diet-induced obese rats, along with the potential benefits of leptin supplementation during suckling. Dams were fed either a standard diet (SD), a western diet (WD) before and during gestation and lactation (WD-dams), or a WD switched to an SD during lactation (Rev-dams). Offspring were supplemented with leptin or vehicle during suckling and then fed an SD or WD until four months. Offspring of the Rev-dams exhibited improved metabolic indicators, including lower body weight, reduced plasma levels of TNF-alpha, a higher adiponectin/leptin (A/L) ratio, enhanced liver antioxidant defenses, and decreased inflammation markers in white adipose tissue (WAT) compared to WD-dams, with sex differences. Leptin supplementation further modulated these markers, reducing oxidative stress in liver and inflammation in WAT and liver (e.g., hepatic Tnfa expression decreased by 45% (males) and 41% (females) in the WD group on an SD), and improving the A/L ratio, with effects varying by maternal conditions and sex. In conclusion, this study underscores the importance of maternal nutrition and leptin intake during suckling in shaping long-term metabolic and inflammatory health in offspring, offering strategies to mitigate the adverse effects of maternal obesity on future generations.

Abstract Early life conditions may influence the gut microbiome and contribute to health status in adulthood. We examined the impact of maternal obesity and/or obesogenic diet on the gut microbiome of mothers (at weaning) and their adult offspring, along with the effects of leptin supplementation during suckling. Three groups of rats were studied: control (C) dams, fed with standard diet (SD); western diet (WD) dams, fed with WD (a high‐fat, high‐sucrose diet) before gestation and during gestation and lactation; and reversion (REV) dams, fed as WD‐dams but with SD during lactation. Offspring were supplemented throughout suckling with leptin/vehicle and weaned on an SD. WD‐dams showed lower microbial diversity and an altered microbiome compared to C‐ and REV‐dams, widely normalized by diet improvement during lactation. WD‐ and REV‐offspring (males) displayed lower microbiome diversity and greater dominance compared to C‐offspring. Taxonomic group differences (Proteobacteria phylum and Bacteroides, Parasutterella, and Phocea genera) were observed in WD‐ and REV‐offspring. WD‐offspring, but not REV‐offspring, had a greater abundance of the phylum Firmicutes and Lachnospiraceae family and a lower abundance of the family Tannerellaceae compared to C‐offspring. Leptin supplementation led to decreased abundance of the phylum Actinobacteria (family Eggerthellaceae and genera Enterorhabdus and Adlercreutzia (females)), and the genera Lachnospiraceae UCG‐008 group, Roseburia and Limosilactobacillus, and increased abundance of the genus Dellaglioa, compared to vehicle‐treated groups. Therefore, maternal consumption of an obesogenic diet during the perinatal period results in microbiome changes in dams, alleviated by dietary improvement during lactation. Maternal conditions and leptin supplementation during suckling modestly influenced the microbiome in adulthood, which might partially contribute to the adverse/beneficial effects, respectively, of these conditions on programmed metabolic health.