Effects Of Ecdysterone On Brown Gene Expression In3t3-L1Cell

Objective: To get the white adipose cell through the induction anddifferentiation of the mouse`s3T3-L1preadipocytes. Then intervene the cell with different concentrations of ecdysterone （ECR）or rosiglitazone（RSG）,observe the changes of groups`metabolic states,and further studythe underlying mechanisms.Methods: Cullture the mouse’s3T3-L1preadipocytes in high glucose DMEM medium,containg10%fetal calf serum. Then choose the cells in the logarithmic growth phase and cultureit in the new culture flasks.When the3T3-L1preadipocytes grow up to70%confluence, cultivate them with high glucose DMEM medium with3-isobutyl-1-methylxanthine（IBMX）0.5mmol/L、dexamethasone1μmmol/L、insulin10μg/ml for2-4days,then cultivate them with high glucose DMEM medium with insulin10μg/ml for2-4days,cultivate them with high glucose DMEM medium for4days.Then incubate the cells withecdysterone or rosiglitazone for24hours.Measure the groups`glucoseconsumption and the changes of cells`triglyceride（TG） rates,and use theRT-PCR and Western blot to determine peroxisome proliferator-activated receptorγ（PPARγ）mRNA and uncoupling protein（UCP1）respectively. The groups are divided into five:（1）normal control group:the3T3-L1adipocyte after the induction and the differentiation （2）ecdysterone group:t he cells are treated with different concentrations of ecdysterone（ECR1×10-7、ECR1×10-6、and ECR1×101×10-5mol/L.）（3） positive control group：the rosiglitazone group with the concentration of1×10-5mmol/L. Results: When the concentration of ecdysterone are greater than1×10-4mmol/L,the OD value decreased significantly.The cells`growth rates are93.70%-100%When the concentration of ecdysterone are between1×10-7₁×10-5.Conclusions:（1） When the concentration of ecdysterone are greater1×10-4mol/L,it is cytotoxic.So the concentration of ecdysterone in thisexperiment are chosen between1×10-7₁×10-5.（2）ECR can increase theglucose consumption of the white adipose cell and decrease its triglyceride（TG） rates.（3）ECR can increase the PPARγexpression in the white adipose cell,increase UP1rates.ECR is likely to “brown” the white adipose cell in this way.