| PurposeTo study the effects of different fatty acid on the gene and protein expression of adiponectin and the gene expression of PPARγ in3T3-L1adipocytes, and observe the possible relationship between adiponectin and peroxisome proliferator activated receptor(PPARγ). It is aim to seek the possible fatty acid or concentration that can up regulating the expression of adiponectin. and the mechanism in3T3-L1adipocytes. It will have great theory value and practical importance to provide the therapeutics or prevention on obesity related disease with original scientific proofs.Methods1. After induced into adipocytes and identified by Oil Red O staining, the3T3-L1adipocytes were treated for palmitic acid(PA), oleic acid(OA), linoleic acid(LA) at different concentration (25、50、100、200、400μmol/L) for24h, and the gene expression of adiponectin mRNA and PPARγ mRNA was determined by real-time PCR(relative quantity).2. And then the3T3-L1adipocytes were treated for palmitic acid(PA), oleic acid(OA), linoleic acid(LA) at concentration of100μmol/L for24h with/without the affect of PPARγ inhibitor GW9662, to determine the gene and protein expression of adiponectin, PPARγ gene expression. Results1. Effects of palmitic acid、oleic acid、linoleic acid on expression of adiponectin mRNA and PPARγ mRNA1.1The mRNA expression of adiponectin and PPARγ in PA group were decreased in a dose-dependent manner, though there was a significant increased at25μmol/L(P<0.05).And at the concentration of400μmol/L to show a significant decreased compared with control(P<0.05). Adiponectin and PPARγ mRNA expression display a significant positive correlation, coefficient r=0.882, P=0.000(P <0.05)1.2The mRNA expression of adiponectin and PPARγ in OA group were increased at25、50、100μmol/L,and both with a significant increased at the concentration of50μmol/L(P<0.05). And then display a descent tendency by a dose-dependent manner with the increased of fatty acid concentration beyond50μmol/L, to a significant at400μmol/L(P<0.05). Adiponectin and PPARγ mRNA expression display a significant positive correlation, coefficient r=0.971, P=0.000(P<0.05)1.3The mRNA expression of adiponectin in LA group were increased at50、100μmol/L(P<0.05),and PPARγ mRNA expression increased at25、50、100μmol/L (P<0.05). At the concentration of400μmol/L significant decrease of both adiponectin and PPARγ mRNA was observed. Adiponectin and PPARγ mRNA expression display a significant positive correlation, coefficient r=0.830, P=0.000(P <0.05)2. Without the effect of PPARγ inhibitor GW9662,100μmol/L PA decreased the expression of Adiponectin (P<0.05) and PPARγ mRNA, but with no effect on the secretion of Adiponectin protein (P>0.05).100μmol/L OA did not affect any of the expression of Adiponectin mRNA, PPARγ mRNA and Adiponectin protein secretion. However, the LA group increased both Adiponectin mRNA,PPARγ mRNA and Adiponectin protein secretion by59%,101%and109%(P<0.05)3. At the meantime, with the effect of PPARγ inhibitor GW9662(100μmol/L), 100μmol/L PA decreased the expression of Adiponectin and PPARγ mRNA, and the secretion of Adiponectin protein significantly (P<0.05). And the OA group with a descent in adiponectin mRNA, PPARγ mRNA expression and Adiponectin protein secretion (P<0.05). The LA group display a significant decrease on both Adiponectin mRNA,PPARγ mRNA and Adiponectin protein secretion compared with control (P<0.001). It is prompting that PPARγ inhibitor GW9662eliminate the up-regulation effect of LA on PPARγ mRNA, and aggravate the depressant effect of PA on PPARγ agonism.Conclusion1. Fatty acid can induce a dose-dependent manner at the gene expression of adiponectin and PPARγ mRNA in3T3-L1adipocytes. Both OA and LA could up-regulate the expression adiponectin at a certain range of concentration. PA can also increase the gene expression of adiponectin and PPARγ at approximately low level of25μmol/L, although it always shows off a depressant effect.2. The molecular mechanism of adiponectin gene expression might be correlated with a PPARγ activation signaling pathway, for there is a positive correlation between adiponectin and PPARγ. And it is prompting that PPARγ inhibitor GW9662eliminate the up-regulation effect of LA on PPARγ, and aggravate the depressant effect of PA on PPARγ agonism. PA,OA,LA may be the possible ligand of PPARγ.3. It is prompting in this study that both category, dose and time are the important factor affecting the expression of adiponectin. And it is possible to educe its beneficial contribution through the very category, dose and time of fatty acids. |
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