The Study Of The Relationship Between Overexpression Of NSREBP-1c And Lipodystrophy

Objective:To observe adipocyte differentiation of brown adipose tissue and white adipose tissue SVF cells from nSREBP-lc overexpression mice in vitro.And to observe whether mice serum from nSREBP-lc overexpression mice affects adipogenesis or lipid metabolism in 3T3-L1 cells and in mice,in order to look for metabolic regulation mechanism of adipogenesis and lipodystrophy.Methods:(1)acquisition and passage cultivation the SVF cells of brown adipose tis sue and white adipose tissue from aP2 nSREBP-lc(aP2)mice which overexpr ess nSREBP-lc in adipose tissue and some generation of wild type(WT)mi ce.and immortalized the cells with SV40 lentivirus with large T antige n.(2)establish preadipocyte cell line from SVF in vitro and induced into mature adipocyte,then by oil red 0 staining,western blot and reverse transcription PCR(RT-PCR)we observe adipogenesis and related genes expr ession.(3)Use aP2 and WT mice serum to induce normal 3T3-L1 cell into ma ture adipocyte,then observe adipogenesis and related genes expression b y oil red 0 staining,western blot and RT-PCR.(4)Use aP2 and WT serum to observe wether aP2 serum effect related genes expression of lipid metabo lism in WT mice.Results:two kinds of SVF cells of brown adipose tissue and white adipose tissue p assage cultivation in stable after affecting by SV40 lentivirus with larg e Tantigen,did not find its differentiation ability recede.(2)SVF cell s induced into mature adipocyte and stainde by oil red 0 found aP2 mice SVF cells better than WT mice SVF cells in adipogenesis(P<0.05);By RT-PCR detection of related genes to observe its adipocyte,we found PPAR g amma and C/EBP alpha mRNA expression in aP2 group of SVF cells is higher than the WT group(P<0.05)after induced into mature adipocyte.western b lot detecting its adipocyte found PPAR gamma protein expression in aP2 gr oup of SVF cells after induced into mature adipocyte is higher than the W T group(P<0.05).(3)preparation aP2 mice and WT mice serum,observe t he cells of normal 3T3-L1 induced into mature adipocyte with these two se rum,by oil red staining observation found that in 3%mice serum groups,a P2 mice serum treatment 3T3-L1 cells express lower PPAR gamma and C/EBP a lpha mRNA and PPAR gamma protein than WT mice serum treatment group(P<0.05).(4)Compared with the WT serum group,the aP2 serum group reduced pp ar-gamma and C/EBP alpha mRNA expression in eWAT(P<0.05).Conclusion:(1)aP2 SVF cells of mice under the condition of in vitro can be induced into mature adipocyte,and there is better adipogenesis than WT SVF cells on the contrary of in vivo.(2)compared with WT mice serum the aP2 mice serum attenuates adipogenesis in 3T3-L1 cells and in WT mice;(3)internal environment(or the serum)of aP2 mice is the key of lipodystrophy and the Disordered differentiation of adipose tissue in aP2 mice.