The Effects Of Arecoline On The Lipid Metabolism Of3T3-L1Adipocytes

Objective(1) To investigate the effects of arecoline on differentiated3T3-L1adipocytes lipidcontent.(2) To investigate the effects of arecoline on the mRNA and protein expression ofLipogenesis lipase FAS in differentiated3T3-L1adipocytes.(3) To investigate the effects of arecoline on the mRNA and protein expression ofhydratase ATGL and HSL in differentiated3T3-L1adipocytes.Methods(1) The3T3-L1preadipocytes were induced to adipocytes with insulin,dexamethone, and IBMX，Observed the form of3T3-L1preadipocytes and adipocytesunder inverted microscopy. Set in the control group (0μmol/L) and the treatmentgroup (25μmol/L,50μmol/L,100μmol/L), after treatment72h, oil red O staininglipid droplet accumulation in the cytoplasm.(2) With different concentrations of arecoline treated cells, total RNA of cells wasextracted in each group, using reverse transcriptase-polymerase chain reaction(RT-PCR) to detect the mRNA expression of lipid metabolism related enzymes ineach group; Western blot to detect the protein expression of lipid metabolism-relatedenzyme.Results(1)3T3-L1preadipocytes were no lipid droplets in cytoplasmic; the differentiatedcells showed a large round, showing a large number of lipid droplets in the cytoplasmsurrounding the nucleus, was "ring ring" shape. Lipid determination showedArecoline mature fat cells to reduce the number of lipid droplets.(2) Arecoline had cytotoxic effect on differentiated3T3-L1adipocytes and it wasconcentration dependent. The cell relative survival decreased with increasingconcentration within the range of200-1600μmol/L. Within0-100μmol/L range, the relative survival rate had no apparent regularity, but overall activity is good. Withdifferent concentrations arecoline treating3T3-L1adipocytes24h,48h,72h and96h,the relative survival rate of treatment time96h was significantly lower than in the24h,48h,72h groups.(2) After the cells were treated with different concentrations of arecoline, theresults show: Arecoline reduce the expression of fatty acid synthase FAS mRNA andprotein, compared with the control group,25μmol/L group arecoline FAS mRNAexpression was reduced by approximately40%, protein expression decreased by15.7%, FASmRNA expression50μmol/L group decreased by t65%, protein expressionwas decreased by approximately46.2%,100μmol/L group FAS mRNA did notchange significantly, but the expression of the protein decreased by27.6%.(3) Arecoline increase the mRNA and protein express of adipocytes triglyceridelipase (ATGL), hormone-sensitive lipase (HSL). Compared with the control group,25μmol/L Arecoline group ATGL mRNA levels increased by47.3%,31.5%increasein protein expression; HSL mRNA levels increased65.4%,43.7%increase in proteinlevels.50μmol/L group ATGL mRNA levels increased by223.2%, and99.2%increase in protein levels; HSL mRNA levels increased by207.8%, and81.8%increase in protein expression levels.100μmol/L group ATGL mRNA expressionlevels increased by198.1%, the protein levels increased by41.6%; HSL mRNAexpression levels increased by112.5%,31.5%increase in protein expression levelsConclusionArecoline can significantly reduce the lipid content of differentiated3T3-L1adipocyte, which may be associated with decreased levels of fatty acid synthaseexpression and increased the express levels of key adipolysis enzyme ATGL andHSL.