The Effects Of Acylation Stimulating Protein On The Glycometabolism And Lipometabolism In3T3-L1Adipocytes

Objective: To evalute the affect on protein expression of classical signaling pathwaysof Insulin (IRS-1, PI3K), and the affect on secretion levels and protein expression ofthe key enzyme of lipid metabolism(LPL,HSL,Perilipin,DGAT) in3T3-L1adipocytes with insulin stimulated state and Acylation stimulatingprotein(ASP)stimulated in different kinds and concentrations of free fatty acids. Toexplore the regulatory of ASP on glucose and lipid metabolism.Method3T3-L1preadipocytes were differentiated to adipocytes by cocktail hormonescontaining10μg/mL insulin,1μmol/L dexamethasone and0.5mmol/Lisobutylmethylxanthine. The morphologic changes of3T3-L1cells were observed bymicroscope. Fat droplets observed in the cytoplasm by Oil red O staining. Thewestern blot were used to detected protein of HSL, Prelipin, IRS-1,PI3K.Realtime-PCR were used to detected mRNA of LPL, HSL, Perilipin, DGAT.Results1.3T3-L1preadipocytes were fibroblastic, and no fat droplet was noted in cytoplasm.After fully differentiated, the adipocytes became bigger, rounder and larges of fatdrops accumulated, which looked like the finger ring.2In insulin and ASP stimulating condition, comparing with the0mmol/L,the levelof HSL, perilipin, PI3K protein expression were increased. And comparing with theinsulin stimulated, the protein expression of HSL,perilipin IRS-1and PISK failed tosignificantly increase with the ASP stimulated.3. After incubate with ASP, comparing with that of0mmol/L, the level of HSL,perilipin, LPL and DGAT mRNA expression were increased. ASP can increase HSL (p<0.05or p<0.01,0.5mmol/L and1.0mmol/L), LPL(p<0.05or p<0.01,0.125、0.5、1.0mmol/L), perilipin (p<0.05or p<0.01,0.125、0.5、1.0mmol/L), DGAT (p<0.05orp<0.01,0.125、0.5、1.0mmol/L).ConclusionIn the condition of insulin resistance, INS and ASP can increase the protein expressionof the key enzyme of lipid metabolism (HSL, perilipin), and the classical insulinsignaling pathway (PI3K). ASP can increase the mRNA expression ofHSL, LPL, perilipin and DGAT. By increasing the expression of these enzymes toimprove the sensitivity and improve the use of the body’s fat cell, to regulate theglucose and lipid metabolism.