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- Yeruva, Laxmi
- Ruebel, Meghan
- 2023
- 2022
- Human Nutrition (107)

Research Project: **Location:
                            Microbiome and Metabolism Research** Project Number: 6026-51000-012-008-S
                        
                        Project Type: Non-Assistance Cooperative Agreement Start Date: Aug 1, 2022
                        
                        End Date: Jul 31, 2027 **Objective:**
                        
                        1. To examine the effects of early life stress on immune response markers in various tissues (i.e., bone, adipose, muscle and heart).
2. Intake of adequate amounts of nutrients including antioxidants during pregnancy and early in life ensures healthy outcomes to both mother and infants. One approach that pregnant mother and infants can take to ensure adequate intake of nutrients and prevent chronic disease later in life is consumption of adequate fruits and vegetables that are rich in antioxidants. Pinto beans (PB, Phaseolus vulgaris L.) are rich in bioactive compounds including resistant starch (RS) and polyphenolic compounds which can act as prebiotic and can confer health benefits during pregnancy and early life. Recent findings in mice from Dr. Lucas's team revealed that PB can counter the effects of a Western-style diet by improving gut integrity, reducing chronic inflammation, and improving insulin sensitivity in adult mice. The knowledge gaps exists in term of beneficial effects of PB diet in an obese condition during pregnancy and in turn programming effects on placenta and offspring. The objective of the study is to investigate PB diet role on the mother and fetal health using high fat diet mouse model. **Approach:**
                        
                        We hypothesized that early life stress during lactation impacts gut health resulting in chronic inflammation that will interfere with optimal function of these tissues.

Amendment #1 -The animal study will be conducted at Oklahoma State University. Four week old female mice (C57Bl/6) will be placed on either control (CON,17% fat, TD95092) or high-fat, high sucrose diet (HFD, 45% calories from fat, TD8811) for 8 weeks to induce obesity. Mice will be switched to either CON, CON + 10% PB, HFD, HFD + 10% PB for 4 weeks with these diets prior to mating with lean control males. Mice will continue on these diets throughout the pregnancy. Fecal samples will be collected once a week throughout gestation for microbiota and metabolomics assessment.  We will collect serum, bones, heart, muscle, placenta, fetus, intestine, cecum, fetal liver and other tissues collection will be conducted at gestation day 18.5 to assess PB diet benefits on dams and placenta.

The animal study will be conducted at USDA-ARS Arkansas Children's Nutrition Center. Pregnant rats will be allowed to deliver pups without intervention. On postnatal day (PND) 2, the litters will be randomly assigned and transferred to either an “impoverished” cage or a standard cage at which point the litter will be left undisturbed and the cage unchanged for 7 days. On postnatal days 2, 10, and 22, we will obtain 1.0 ml milk sample from each dam at the time of planned separation from the dam. Postnatal food consumption will be measured, but not restricted. We will weigh the food provided during routine cage changes and daily animal checks. Dams and pups will be weighed at the time of routine cage changes.

To determine how early life stress during lactation impacts bone health, the Collaborator's team will measure bone quality (bone mineral content, bone mineral density, and microarchitecture) and gene expression of inflammatory markers. Furthermore, adipose tissue- gene expression (inflammatory markers-IL6, TNF-alpha, uncoupling protein, glucocorticoid receptor, leptin, glucose transporter- GLUT4), histological examination (adipocyte size & lipid density), muscle - gene expression (inflammatory markers, glucose transporter GLUT 4) and heart and aorta - gene expression of inflammatory and vascular dysfunction markers will be measured. ARS SY's team will assess gut microbiota composition to associate the bone, muscle, heart, and adipose tissue outcomes.


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