AGX0104 Can Block The Formation Of Fat And Insulin Sensitivity During Adipogenesis

Evidence from the recent studies indicates that insulin resistant and Adipogenesis are directly associated with an increased risk of Endometrial Carcinoma. During the Adipogenesis and the development of insulin resistant: two important transcription factors play an important role. One is PPARs(members of the nuclear hormone receptor surperfamily), another one is C/EBPs. It seems that mechaniams that control expression of PPARs and C/EBPs are likely to influence the Endometrial Carcinoma, but how they work and do they have the relationship during Adipogenesis and the development of insulin resistant, it is still unkown. In the present study , we set out to define the role of each of these factors during Adipogenesis and insulin resistant. One strategy involved the use of a potent PPAR gammer selected antagonistAGXO104) to block PPAR gammer activity. Treatment of differentiating preAdipocytes with PPAR gammer antagonist inhibits C/EBP a expression and Adipogenesis at a step downsteam of PPAR gammer, suggesting that the expression of C/EBP a depends on PPAR y avtivity. To Address the contribution of C/EBP beta and PPAR gammer to the activation of C/EBP a transcription , we ectopically expressed C/EBP beta in Swiss MEFs using a TET-OFF conditional expression system. Overexpression of C/EBP P in the presence of the Adipogenic inducers: dexamethasone, insulin and iaobutylmethylxanthine activated PPAR y 2 expression, but not C/EBP a expression. Taken together, these data suggest that induction of Adipogenesis requires C/EBP beta to induce PPAR y, which then may activates C/EBP a expression .Consequently , effectors which regulate C/EBPbeta as well as PPAR gammer, C/EBPa activity are potential regulators of Adipogenesis and insulin sensitivity, thus inhibit the formation of Endometrial Carcinoma. So AGX0104 is an effective compound to block the formation and the development of fat and insulin sensitivity during Adipogenesis.Methods:1. Subclone the interested gene into Pbig-Puro vector.2. Transformation.3. Transfection, make stable C/EBP p expression cell line.4. Using Western to measure the expression of PPARy , C/EBP p ,C/EBP a , Adiponectin, Perilipin.Results:1. AGX0104 can block the expression of PPARy and C/EBPa and Adiponectin and Perilipin during Adipogenesis of 3T3-L1 cell line, but it can not block the expression of C/EBP p.2. During the Adipogenesis of 3T3-L1, AGX0104 can totally block the expression of PPARy and Adiponectin(the factor that can measure the insulin sensitivity), so does perilipin (the factor that can measure the development of lipid drop) .3. The expression of C/EBP p can induce Adipogenesis , but it can not get the expression of PPAR y receptor. PPAR y receptor can activate PPAR y ,thus induce the expression of C/EBP a.4. Ectopically expressed C/EBP beta in swiss MEFs using a TET-OFF conditional expression system. Overexpression of C/EBP beta in the presence of the Adipogenic inducers dexamethasone , insulin and iaobutylmethylxanthine activated PPAR y 2 and perilipin expression ,but not C/EBP a and Adiponectin expression .But if we use the dose response of TET , we found -.with the decrease expression of C/EBP P , the expression of PPAR y and C/EBP a also decreased. This means C/EBP p locates the upstream of the signal transduction of PPAR y and C/EBP a .5. During the Adipogenesis of C/EBP p cells , after the inducement ,we can get the expression of PPAR y shortly .But we can not measure the expression of C/EBP a. Atday 2 (after induce),Add PPARy ligand, the expression of C/EBP a is activated, with the expression of C/EBPa ,the cells can express Adiponectin , without the expression of C/EBPa , the cells can not express Adiponectin,even with the expression of PPARy. with the expression of PPARy,the cells can express perilipin ,even without the expression of C/EBPa. C/EBP P is an important effector .After the expression of PPAR y is activate , C/EBP P will not be the important factor any more .It began to degrAdation in day 6.Conclusion:1.