Banca de DEFESA: ÂNGELA IMPERIANO DA CONCEIÇÃO
Uma banca de DEFESA de DOUTORADO foi cadastrada pelo programa.STUDENT : ÂNGELA IMPERIANO DA CONCEIÇÃO
DATE: 26/02/2024
TIME: 09:30
LOCAL: UFRPE
TITLE:
GENE EXPRESSION OF mRNA FOR ADIPOKINS AND ITS RELATIONSHIP WITH METABOLIC DYNAMICS IN COWS WITH AND WITHOUT HYPERKETONEMIA
KEY WORDS:
Keywords: ketosis; leptin; adiponectin; real-time PCR; multivariate analysis; molecular biology; negative energy balance.
PAGES: 114
BIG AREA: Ciências Agrárias
AREA: Medicina Veterinária
SUBÁREA: Clínica e Cirurgia Animal
SPECIALTY: Clínica Veterinária
SUMMARY:
Understanding the physiological roles of adipokines in ruminants is crucial to improving animal health and productivity, as they are intrinsically related to the states of metabolic regulations, energy balance, glucose and lipid metabolism, inflammatory response, endocrine, affecting sensitivity insulin and metabolic syndromes. Therefore, the objective was to verify the expression of mRNA for leptin, leptin receptor (ObRb), adiponectin, adiponectin receptor (AdipoR1) and resistin in placental tissue of cows with and without hyperketonemia and to evaluate their interaction with the energetic metabolic profile, protein, enzymatic, hormonal and mineral. To this end, 135 pregnant cows were used and sent for clinical-obstetric care at the Garanhuns Bovine Clinic/UFRPE. The samples were collected at the time of birth and the females were subsequently distributed into groups, whose inclusion indicators for the cows were: G1 without hyperketonemia (n=120) (β-Hydroxybutyrate <1.2mmol/L) and G2 with hyperketonemia (n=15) (β-Hydroxybutyrate ≥1.2mmol/L). The biochemical indicators determined in the blood were: NEFA, β-Hydroxybutyrate, glucose, total protein, albumin, urea, creatinine, GGT, AST, chlorides, phosphorus, ionizable calcium, potassium and sodium ions and the hormones insulin and cortisol. Insulin sensitivity was assessed using mathematical models (HOMA-IR, QUICKI, RQUICKI and RQUICKIΒHB). Fragments of each placenta were collected and stored in cryotubes containing Trizol and kept at -80°C. Of the 135 placenta samples collected from cows, 38 were processed and analyzed, 23 from G1 and 15 from G2. mRNA extraction was performed using the trizol method. cDNA was synthesized from 1μg of total RNA. qPCR analyzes were performed in duplicates. The difference between the mean Ct of the gene of interest and the mean Ct of the endogenous gene (ΔCT) was calculated for normalization and the result for gene expression was determined by a dimensionless value using the formula 2-ΔΔCt. The data was processed using PROC MIXED from SAS. The variables were subjected to the normality test to verify whether or not they met the normality assumptions using different tests, such as the Shapiro-Wilk, Kolmogorov-Smirnov and Anderson-Darling tests. Data that did not meet these assumptions were subjected to transformation, using a logarithmic or radical basis. For the set of variables with a parametric profile, the F Test was performed, followed by a mean study using the SNK Test. For the set of variables indicating the application of a non-parametric test, the data were submitted to the Mann-Whitney U Test. For all statistical analyzes carried out, a significance level (p) of 5% will be adopted. For multivariate analysis, pre-processing was carried out considering: 1. Imputation of missing data; 2. SMOTE: Synthetic sampling in the case (after sample selection) and 3. Auto scaling: Different variances per variable. Therefore, different models were carried out: 1. Exploratory analysis and 2. Supervised pattern recognition. In the present work, statistical techniques of multivariate analysis were used, applying PCA (Principal Component Analysis), HCA (Hierarchical Component Analysis) and PLS (Partial Least Squares), seeking to evaluate the best response in the analyzes carried out. Serum NEFA concentrations were marked in the group of animals with hyperketonemia (p=0.0002). Glucose concentrations were significantly lower in cows with hyperketonemia (p=0.0132), as were the percentages of RQUICKIβHB (p<.0001). Serum urea concentrations were significantly higher in G2 females. Hyperketonemic cows had higher mean values for leptin (p=0.0174) and ObRb (<.0001) expression and healthy cows had higher mean values for adipoR1 and resistin expressions (<.0001). In the analysis of the relationship between pairs of variables, it was found that the variables with the greatest capacity for discrimination between the groups studied were the expression of ObRb, expression of AdipoR1, β-Hydroxybutyrate, AGNE, glucose, GGT and RQUICKBHB, in addition to having varying degrees relationship, highlighting: positive correlation of leptin expression with the expression of ObRb, β-Hydroxybutyrate; positive correlation of ObRb expression with β-Hydroxybutyrate, NEFA, GGT, AST and negative with glucose; positive correlation of AdipoR1 expression with glucose, RQUICK and RQUICKBHB, and negative with β-Hydroxybutyrate, NEFA and GGT; negative correlation of β-Hydroxybutyrate and NEFA with RQUICK and RQUICKβHB. In HCA, it is evidenced that AdipoR1 and β-Hydroxybutyrate play a crucial role in differentiating the groups and identifying distinct metabolic patterns associated with hyperketonemia; in PCA, AdipoR1 and RQUICKBHB emerged as key variables for the control group, while AGNE and β-Hydroxybutyrate stood out for the hyperketonemia group. Regarding PLS for the hyperketonemia case class, it was identified that the expression of ObRb, expression of AdipoR1, and β-Hydroxybutyrate are representative of the hyperketonemia condition in cows at the time of calving. The analysis of adipokines produced in the placenta is a new tool to understand the role of these molecules in the mechanisms regulating metabolism in cows. The increase in leptin and ObRb gene expression and the reduction in AdipoR1 gene expression in hyperketonemic cows and the changes in metabolism and associated biochemical processes stand out, showing that the placental production of this hormone can have an important contribution to metabolic conditions, such as hyperketonemia, bringing implications for the physiology, health and productivity of females.
COMMITTEE MEMBERS:
Presidente - PIERRE CASTRO SOARES
Interno - ***.333.884-** - JOSE AUGUSTO BASTOS AFONSO DA SILVA - UFRPE
Externo ao Programa - 1331645 - PAULO ROBERTO ELEUTERIO DE SOUZA - UFRPEExterno ao Programa - 2426003 - RODOLFO ARAUJO DE MORAES FILHO - UFRPEExterna à Instituição - MARIA CLAUDIA ARARIPE SUCUPIRA