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Effects of Insulin on Adiponectin and AdipoRs mRNA Expression in Cultured Bovine Adipocytes

Author: SunYongGang
Tutor: ZuoLinSen
School: Northwest University of Science and Technology
Course: Cell Biology
Keywords: Bovine fat cells Insulin Adiponectin Adiponectin Receptors PI3K
CLC: Q813
Type: Master's thesis
Year: 2009
Downloads: 56
Quote: 0
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In this study, newborn Holstein bull calves as experimental material collected aseptically inguinal and epididymal adipose tissue, with type I collagenase digestion and tissue culture were isolated and cultured bovine preadipocytes by fat cells in the culture morphology, growth curve drawing, insulin-induced differentiation, oil red O staining and preadipocytes and adipocytes marker gene detection, build of bovine preadipocytes vitro proliferation and differentiation model. Detection of adiponectin, of AdipoR1, and AdipoR2, Pref-1 and PPAR-gamma mRNA expression in bovine preadipocytes process law. The same time, fat cell differentiation 9 d objects of different concentrations and different processing time, the impact of insulin on adiponectin, AdipoR1 and AdipoR2 mRNA expression and may affect the metabolic pathways. The study reveals cattle proliferation and differentiation of preadipocytes law to explore the possible regulatory mechanism in the adiponectin receptor gene in cattle fat cells during proliferation and differentiation, and laid the foundation for improving our local cattle intramuscular fat content. The main findings are as follows: initially built the proliferation and differentiation of bovine preadipocytes in vitro model with type I collagenase digestion can be separated from the adipose tissue of newborn calf large number of preadipocytes, insulin-induced differentiation, oil red O staining and preadipocytes cell marker gene Pref-1 and induction of fat cell marker gene PPAR-gamma detection, in vitro cultured preadipocytes confluence 9 d differentiate into fat cells. 2. Clarify the adiponectin, AdipoRs, Pref-1 and PPAR-gamma expression pattern of Adiponectin in cattle, fat cell differentiation process in cattle before the fat cells do not express significant increase with preadipocytes induced differentiation adiponectin and AdipoR2 mRNA expression (P lt; 0.05), AdipoR1 sustained expression in the process of cell differentiation and expression levels was not significant (P gt; 0.05) Pref-1 mRNA high expression of the first fat cells was significantly reduced with the differentiation of fat cells, and not expressed in the the 8 d differentiation of cells in the same time, in the differentiated cells Pref-1 expression was significantly lower than the normal cultured cells (P lt ; 0.05). PPAR-gamma expression increased with the differentiation of preadipocytes and differentiated cells was significantly higher than control cells (P lt; 0.05). Study different insulin concentration and treatment time on the adiponectin receptor gene expression changes induced differentiation of the 9 d cattle fat cells in different treatment concentrations of insulin inhibited adiponectin mRNA expression and dose-dependent inhibition, while , 100 nmol / L was time-dependent inhibition of the expression of adiponectin. Different concentrations of insulin can suppress the expression of AdipoR2 and the inhibitory effect of each dose difference was not significant, and 100 nmol / L insulin for 24 h could significantly inhibit the expression of AdipoR2. Different concentrations of insulin and insulin processing time AdipoR1 expression. Initially revealed the bovine fat cells in insulin through PI3K/Akt signaling pathway of adiponectin and AdipoR2 transcription of, and suppression is reversible.

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