Effect of overexpression of Fad24 on intracellular Dex receptor and auxiliary mo-lecular expression in porcine DFAT cells under lipid-forming induction conditions
The aim of this experiment was to investigate the effect of overexpression of Fad24 on the expression of intracellular dexamethasone(Dex)receptor GR and its high-affinity helper mole-cules Hsp90 and FKBPs in porcine dedifferentiated adipocytes(DFAT)under lipogenic induction conditions.Porcine DFAT cells were transiently transfected with overexpression vector pcDNA-Fad24 and empty vector pcDNA,and lipogenic differentiation was induced in a high concentration of Dex(1 μmol/L)lipogenic induction solution,and then the induced differentiation was examined according to morphological observations,and the effects on GR and the expression of Hsp90 and FKBPs were examined by real-time PCR and Western blot techniques.The results showed that the pcDNA empty vector group could induce lipogenic cells after 5 d of induction,while the pcDNA-Fad24 overexpression group completely inhibited lipogenic differentiation,with cells accumulating to form multiple small nodules and forming typical osteogenic nodules after 7 d of induction,and a-lizarin red staining confirmed the deposition of mineralized material on the nodule surface.The expression of GRα mRNA was significantly up-regulated at 48 h of lipogenesis induction(P＜0.01),while that of GRβ mRNA was significantly up-regulated at 12 h of lipogenesis induction(P＜0.01);the expression of Hsp90 mRNA,a high affinity helper molecule of GR,was significant-ly up-regulated at 48 h of lipogenesis induction(P＜0.01);the expression of FKBP 51 mRNA was significantly up-regulated at 24 h of lipogenesis induction.The expression of FKBP 51 mRNA was significantly upregulated at 24 h of lipogenesis induction(P＜0.01),while the expression of FKBP 52 mRNA was significantly upregulated at 48 h of lipogenesis induction(P＜0.01).The effect of Fad24 overexpression on the transformation of porcine DFAT cells from lipogenic to osteogenic differentiation was associated with the upregulation of GRα and its Hsp90 and FKBP 52 expression at 48 h of lipogenic induction,and the downregulation of GRβ and FKBP 51 expression.
overexpression of Fad24DFAT cellsinduction of lipid formationglucocorticoids recep-torhigh affinity helper molecule
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