ABSTRACT
BACKGROUND Gestational protein restriction was associated with low birth weight, hypertension and higher prevalence of cardiac disorders in adults. Several mechanisms, including epigenetics, could be related with the cardiovascular phenotype on protein-restricted offspring. Thus, we investigated the morphological cardiac effects of gestational protein restriction and left ventricle miRNAs and target genes expression pattern in both 12-day and 16-week old gestational protein-restricted male offspring.
METHODS Pregnant Wistar rats were allocated into two groups, according to protein supply during pregnancy: NP (normal protein diet-17%) or LP (low protein diet − 6%). The study evaluates the effects of maternal protein restriction on food consumption and body weight of both pregnant dams and offspring, systolic blood pressure in 16-wk old offspring and on cardiac morphometric and molecular parameters in both 12-d and 16-wk old offspring.
RESULTS Dams on the gestational protein-restricted diet had lower body weight gain and higher food intake. Gestational protein-restricted offspring had low birth weight, followed by rapidly body weight recovery, hypertension, and increased myocytes crosssectional area and collagen fraction at 16-week old age. At 12-days old, miR-184, miR-192, miR-376c, miR-380-3p, miR-380-5p, miR-451, and miR-582-3p had increased expression, and miR-547 and miR-743 a had decreased expression in the gestational protein-restricted left ventricle. At 16-week old, let-7b, miR-125a-3p, miR-142-3p, miR-182 and miR-188-5p had increased expression and let-7g, miR-107, miR-127, miR-181a, miR-181c, miR-184, miR-324-5p, miR-383, miR-423-5p and miR-484 had decreased expression in gestational protein-restricted left ventricle. Target predicted gene expression analysis shown higher expression of Dnmt3a, Oxct1, Rictor and Trps1 and lower expression of Bbs1 and Calml3 in 12-day old protein-restricted offspring. 16-week old protein-restricted offspring had higher expression of Adrbk1, Bbs1, Dnmt3a, Gpr22, Inppl1, and Oxct1 genes.
CONCLUSION Gestational protein restriction leads to offspring low birth weight, increased systolic blood pressure and morphological heart alterations that could be related to early heart miRNA expression changes that perpetuate into adulthood and which are associated with the regulation of essential genes involved in cardiovascular development, heart morphology, function, and metabolism.
Authors’ contributions
HBA: Data curation, Investigation, Formal analysis, Methodology, Visualization, Writing-original draft; JARG: Formal analysis, Methodology, Visualization, Writing-review & editing; PAB: Conceptualization, Formal analysis, Funding acquisition, Methodology, Resources, Supervision, Visualization, Writing-original draft, Writing-review & editing
Acknowledgments
The authors thank Dr. Tom Fleming - University of Southampton for the collaboration with the paper review.
- List of abbreviations
- 11β-HSD
- 11beta-hydroxysteroid dehydrogenase
- 2-way ANOVA
- two-way analysis of variance
- Adrbk1 gene
- Beta adrenergic receptor kinase 1 gene
- Bbs1 gene
- Bardet-Biedl Syndrome 1 gene
- Calml3 gene
- Calmodulin-like protein 3 gene
- cDNA
- complementary DNA
- COBEA
- Brazilian College of Animal Experimentation
- CSA
- cross-sectional area
- DNA
- deoxyribonucleic acid
- Dnmt3a gene
- DNA methyltransferase 3 alpha gene
- GAPDH
- glyceraldehyde 3-phosphate dehydrogenase
- Glut4
- type 4 glucose transporter
- Gpr22 gene
- G-protein coupled receptor 22 gene
- HE
- hematoxylin and eosin
- IDT®
- Integrated DNA Technologies
- Inppl1 gene
- Inositol Polyphosphate Phosphatase Like 1 gene
- LP
- low protein diet
- LV
- left ventricle
- miRNAs or mir-or miR
- –MicroRNAs (miR-mature form of the miRNA; mir-pre-miRNA)
- mRNA
- messenger RNA
- NIH
- National Institutes of Health
- NP
- normal protein diet
- Oxct1 gene
- 3-Oxoacid CoA-Transferase 1 gene
- PI3K-Akt
- phosphatidylinositol 3’-kinase (PI3K)-Akt signaling pathway
- Rictor gene
- Rapamycin-insensitive companion of mammalian target of rapamycin gene
- RNA
- ribonucleic acid
- Ship2
- SH2 domain containing inositol 5-phosphatase 2
- Trps1 gene
- Transcriptional Repressor GATA Binding 1 gene
- UNICAMP–
- State University of Campinas
Footnotes
‘Declarations’
‘Declarations’ Ethics approval and consent to participate: The Institutional Ethics Committee (CEU A/UNICAMP #315) approved the experimental protocol, and the general research guidelines established by the Brazilian College of Animal Experimentation (COBEA) and by NIH Guide for the Care and Use of Laboratory Animals were followed throughout the investigation. The experiments were conducted as described in detail previously [49,50] on age-matched rats of 12-week-old sibling-mated Wistar HanUnib rats (250-300 g). The local colonies originated from a breeding stock supplied by the Multidisciplinary Center for Biological Investigation on Laboratory Animal Science, UNICAMP, Brazil.
Availability of data and material: Available in http://repositorio.unicamp.br/handle/REPOSIP/322389 Moreover, https://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&idtrabalho=3646388
Competing interests: No conflicts of interest, financial or otherwise, are declared by the authors.
Funding: This work was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (2013/12486-5); Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES; 9128/12-3); and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).
Consent for publication: Not applicable.