Screening Study On Activity Of Ameliorating Insulin Resistance In Vitro Of Berberine Hydrochloride Structure Modification Based On 3T3-L1 Pre-adipocytes

Objective:Exploratory research on methods of 3T3-L1 pre-adipocytes differentiation and insulin resistance establishment:In order to find scientific and rational methods of screening multitude activity of ameliorating insulin resistance.Meanwhile,study on the affection to ameliorating insulin resistance of berberine hydrochloride structure modifications.Methods:1. Determine 3T3-L1 pre-adipocytes proliferation in 84 hours by MTT.2. Optimize the method of 3T3-L1 pre-adipocytes differentiation by uniform design.3. Research the affection to 3T3-L1 pre-adipocytes insulin resistance establishment of insulin for scientific and rational methods.4. Based on the studies above,investigate the affection to ameliorating insulin resistance of 38 berberine hydrochloride structure modificationsResults:1. After 48 hours,cells proliferated to the contact inhibition,and stopped to grow obviously.It shows that it is the best moment to differentiate.2. According to the regression equation, obtained the most efficacious method of differentiation:added the differentiated culture medium when cells proliferated to the contact inhibition including 1μmol/L dexamethasone,0.5mmol/L IBMX and 100μg/L insulin.48 hours later,only kept 1μg/L insulin in the culture edium for another 48 hours.At last use the normal culture medium for 13 days totally.3. Result shows that added both insulin and dexamethasone can establish better insulin resistance mold of adipocytes.Therefore, we use the insulin resistance culture medium including 1μmol/L dexamethasone and 1μg/ml insulin.4. Three times screening results show that samples of FB-48,FB-50,FB-79,FB-38,FB-41 obviously promoted glucose utilization of adipocytes (p<0.05 or p<0.01 Conclusion:1. Obtained scientific and rational methods of 3T3-L1 pre-adipocytes differentiation and insulin resistance establishment.2. Part of samples obviously promoted glucose utilization of adipocytes (p<0.05 or p<0.01),and showed better effective than berberine hydrochloride.These structure modifications are valuable to further study.