Mechanism Research Of Piperine Improve Insulin Resistance Model In3T3-L1Adipocytes

Object:Previous study proved that ehtanol extract of piper longu L.could regulate fastingblood glucose levels and oral glucose tolerance index refiects the level of InsulinResistance of high glucose and high fat diet induced insulin resistance model rats.Based on the previous research, this research subject with piper longum L. main activecomponents of piperine and insulin resistance model commonly used3T3-L1adipocytes as the research object, conducted the following research:1The identification of3T3-L1adipocytes and3T3-L1adipocytes insulinresistance model is set up.2Explore piperine on3T3-L1adipocytes insulin resistance model of intervent-ion and may be involved in the related factor of change.Methods:1Insulin Resistance modelIn order to make the3T3-L1adipocytes differentiation into mature fat cells in ashort time, we use two differentiation fluid interactions and through the detection ofintracellular triglyceride content of triglyceride the mature cell were identified.2Piperine on3T3-L1adipocytes insulin resistanc model and the related indicator-s of detectionTo determine the proliferative effect of piperine on the normal3T3-L1adipocyt-es by MTT assay; with cortical hormone stimulation method to establish3T3-L1adipocytes insulin resistance model,observation of piperine and positive drug, rosiglit-azone and AICAR) for insulin resistance adipocytes of the consumption of glucoseand free fatty acid (FFA) content,the effects of piperine compared with positive drug effect on the improvement of the insulin resistance.3Piperine on3T3-L1adipocytes insulin resistanc model possible molecularmechanism of the effectThe phosphorylated adenosine monophosphate activated protein kinase(pAMPK)and acetyl-CoA carboxylase synthetase(ACC) were detected by enzyme linkedimmunoas-say; HPLC detection cell within a adenosine monophosphate (AMP/ATP)and adenosine triphosphate ratio, so that we can better observe whether piperine is byinfluencing the AMPK pathway play the role of improving insulin resistance.Result:1Differentiation of3T3-L1adipocytes "ring" of lipid droplets can be dyed redby oil red O; meanwhile detection of intracellular triglyceride levels, differentiatedgroup was obviously higher than normal control group.1μM dexamethasone induced24h,48h,72h, compared with normal control group both can reduce the consumptionof glucose.1μM dexamethasone induced48h is the best time that insulin resistancemodel is set up.2The inhibitory concentration50%of piperine in3T3-L1adipocytes is118.3μM, and piperine concentration under15μM on cell proliferation is not too muchinfluence; piperine in5~50μM cell concentration can significantly increase the insulinresistance model on glucose consumption, piperine effect on insulin resistance modelto improve the median effective concentration of6.3μM; compared with model group,piperine, rosiglitazone (ROG) and AICAR cells can significantly increase the insulinresistance model on the consumption of glucose, one of piperine effect is most obviou-s when50μM (P<0.001), compared with model group, piperine, ROG and AICARcan make insulin resistance model intracellular FFA levels decreased (P<0.05).3Compared with model group, piperine, ROG and AICAR group of intracellularpAMPK level increased significantly, the ACC level decreased obviously, comparedwith ROG group and AICAR, piperine group pAMPK level no difference in24h,48hbelow ROG and AICAR, ACC level no difference.Compared with model group,piperine,ROG AMP/ATP ratio in cells increased significantly,AICAR group AMP/ ATP ratio is no difference in the cell, piperine AMP/ATP ratio is slightly lower than inthe cell group ROG group.Conclusion:Insulin resistance usually caused adipocytes of sugar consumption ability, andmade it produce more of the free fatty acids; free fatty acids caused fatty toxicity andwill further aggravate cell insulin resistance. Piperine would increase insulinresistance in adipocytes sugar consumption ability, reduce the free fatty acids in thecells, the effect is similar to popular diabetes drug rosiglitazone. Piperine treatmentled to the decrease of the ATP level of insulin resistance in adipocytes, made thepAMPK activation, speed limited further inhibit fatty acid synthesis enzymeexpression of ACC, that might be the main signaling pathways involved in thepiperine improve insulin resistance.