Effects of diisononyl phthalate on lipid metabolism in HepG2 cells
[Background]Exposure to diisononyl phthalate(DINP),an endocrine disruptor associated with metabolic diseases and widely used in plastic products,has been linked to the development of several adverse health outcomes in the liver,including non-alcoholic fatty liver disease(NAFLD).[Objective]To investigate the effects and the possible molecular mechanisms of DINP exposure on lipid metabolism in human hepatocellular carcinoma cells(HepG2 cells).[Methods]First,HepG2 cells were treated with DINP at three time spots(24,48,and 72 h)and eleven doses(0,0.003,0.01,0.03,0.1,0.3,1,3,10,30,and 100 mmol·L-1).Cell viability were de-tected using cell counting kit 8(CCK8).Intracellular lipid deposition was determined by oil red O staining and lipid content detection,and triglyceride(TG)and cholesterol(TC)were further de-tected.Finally,the mRNA expression levels were detected by fluorescence quantitative PCR,in-cluding fatty acid synthesis related genes[acetyl-CoA carboxylase alpha(Accα),fatty acid synthase(Fasn),malonyl-CoA decarboxylase(Mlycd),and sterol regulatory element binding protein 1(Srebp1)]and β-oxidation related genes[peroxisome proliferator activated receptor alpha(Pparα),AMP-activated protein kinase(Ampk),carnitine palmitoyltransferase 1A(Cpt-1a),transcription factor A,mitochondrial(Tfam),nuclear respiratory factor 1(Nrf1),and peroxisome proliferator-activated receptor gamma and coactivator 1 alpha(Pgc1-α)].[Results]Compared with the control group(0 mmol·L-1),the no observed adverse effect levels(NOAEL)of HepG2 cell viability were 0.3,0.1,and 0.1 mmol·L-1 after 24,48,and 72 h exposure to DINP,respectively,and the corresponding lowest observed adverse effect levels(LOAEL)were 1,0.3,and 0.3 mmol·L-1,respectively(P<0.05).After exposure to 30 mmol·L-1 and 100 mmol·L-1 DINP for 24 h,the intracellular lipid content,lipid deposition,TG,and TC levels were increased significantly compared with the control group(P<0.01).Compared with the control group,the mRNA expression levels of genes related to fatty acid synthesis,such as Mlycd,Srebp1,Fasn,and Accα,were down-regulated after the 100 mmol·L-1 DINP exposure for 24 h,while the mRNA expression level of Mlycd was up-regulated in the 30 mmol·L-1 group.The β-oxidation related genes such as Ampk,Pparα,and Tfam were up-regulated significantly after the 100 mmol·L-1 DINP expo-sure,while Cpt-1a mRNA expression level was down-regulated(P<0.05).[Conclusion]Exposure to DINP at 30 mmol·L-1 and 100 mmol·L-1 can interfere with fatty acid synthesis and β-oxidation in lipid metabolism of HepG2 cells,resulting in lipid deposition.
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