RESUMO
Adipose tissue (AT) expands through both hyperplasia and hypertrophy. During adipogenesis, adipose stromal and progenitor cells (ASPCs) proliferate and then accumulate lipids, influenced by the local AT microenvironment. Increased adipogenic capacity is desirable as it relates to metabolic health, especially in transition dairy cows where excess free fatty acids in circulation can compromise metabolic and immune health. Our aim was to elucidate the depot-specific adipogenic capacity and ECM properties of subcutaneous (SAT) and visceral (VAT) AT of dairy cows and define how the ECM affects adipogenesis. Flank SAT and omental VAT samples were collected from dairy cows in a local abattoir. Tissue samples were utilized for transcriptome analysis, targeted RT-qPCR for adipogenic markers, adipocyte sizing, assessment of viscoelastic properties and collagen accumulation, and then decellularized for native ECM isolation. For in vitro analyses, SAT and VAT samples were digested via collagenase, and ASPCs cultured for metabolic analysis. Adipogenic capacity was assessed by adipocyte size, quantification of ASPCs in stromal vascular fraction (SVF) via flow cytometry, and gene expression of adipogenic markers. In addition, functional assays including lipolysis and glucose uptake were performed to further characterize SAT and VAT adipocyte metabolic function. Data were analyzed using SAS (version 9.4; SAS institute Inc., Cary, NC) and GraphPad Prism 9. Subcutaneous AT adipogenic capacity was greater than VAT's, as indicated by increased ASPCs abundance, increased magnitude of adipocyte ADIPOQ and FASN expression during differentiation, and higher adipocyte lipid accumulation as shown by an increased proportion of larger adipocytes and abundance of lipid droplets. Rheologic analysis revealed that VAT is stiffer than SAT, which led us to hypothesize that differences between SAT and VAT adipogenic capacity were partly mediated by depot-specific ECM microenvironment. Thus, we studied depot-specific ECM-adipocyte crosstalk using a 3D model with native ECM (decellularized AT). Subcutaneous AT and VAT ASPCs were cultured and differentiated into adipocytes within depot-matched and mis-matched ECM for 14d, followed by ADIPOQ expression analysis. Visceral AT ECM impaired ADIPOQ expression in SAT cells. Our results demonstrate that SAT is more adipogenic than VAT and suggest that divergences between SAT and VAT adipogenesis are partially mediated by the depot-specific ECM microenvironment.
RESUMO
The objective of our study was to evaluate the effects of timing of palmitic acid (C16:0) supplementation during early lactation on nutrient digestibility, energy intake and balance, and metabolic responses of dairy cows. Fifty-two multiparous cows were used in a randomized complete block design experiment. During the fresh (FR) period (1-24 d in milk) cows were assigned to either a control diet containing no supplemental fat (CON) or a C16:0-supplemented diet [PA; 1.5% of diet dry matter (DM)]. During the peak (PK) period (25-67 d in milk) cows were assigned to either a CON diet or a PA diet (1.5% of diet DM) in a 2 × 2 factorial arrangement of treatments considering the diet that they received during the FR period. During the FR period, compared with CON, PA increased DM digestibility by 3.0 percentage units and neutral detergent fiber (NDF) digestibility by 4.4 percentage units, and the increase in these variables was consistent over time. Although PA did not affect 18-carbon fatty acid (FA) digestibility, it decreased 16-carbon FA digestibility by 10.8 percentage units and total FA digestibility by 4.7 percentage units compared with CON. We observed a tendency for an interaction between treatment and time for total FA digestibility and 16-carbon FA digestibility due to the difference in FA digestibility between PA and CON reducing over time. Compared with CON, PA increased digestible energy intake by 3.9 Mcal/d, metabolizable energy intake by 3.5 Mcal/d, and net energy for lactation intake by 2.5 Mcal/d. The PA diet also increased milk energy output, negative energy balance, and plasma nonesterified fatty acid concentration and reduced plasma insulin concentration. We also observed a tendency for an interaction between treatment and time for energy balance due to cows receiving the PA treatment being in a greater negative energy balance over time. During the PK period, PA increased DM digestibility by 2.9 percentage units and NDF digestibility by 3.5 percentage units compared with CON. Although PA decreased 16-carbon FA digestibility by 7.0 percentage units, PA did not affect 18-carbon FA digestibility or total FA digestibility. Feeding PA during the PK period increased energy intake and milk energy output and did not affect energy balance. In conclusion, feeding a C16:0 supplement to early-lactation cows consistently increased DM and NDF digestibilities and energy intake compared with a control diet containing no supplemental fat. Feeding C16:0 markedly increased milk energy output in both the FR and PK periods but increased negative energy balance only in the FR period.