SIRT3在脱氧胆酸诱导人正常结肠上皮NCM460细胞能量代谢损伤中的作用

Role of SIRT3 in dysfunction of energy metabolism induced by deoxycholic acid in human colon NCM460 cells

目的:探讨脱氧胆酸(deoxycholic acid,DCA)对人结肠上皮NCM460细胞能量代谢损伤的影响及可能机制。方法:培养人正常结肠上皮NCM460细胞,采用DCA(10μmol/L、30μmol/L和100μmol/L)作用5 d,或DCA(100μmol/L)作用3、5和7 d;DCA(100μmol/L)处理3 d后,用去乙酰化酶sirtuin 3(SIRT3)激动剂白藜芦醇(resveratrol,REV)处理细胞后继续培养至第7天。细胞腺苷三磷酸(adenosine triphosphate,ATP)和乳酸生成量用试剂盒检测,Western blot检测细胞的SIRT3表达。结果:DCA能浓度和时间依赖性地抑制NCM460细胞生成ATP并促进乳酸生成,抑制SIRT3蛋白表达;白藜芦醇能逆转DCA对NCM460细胞的作用。结论:DCA对人结肠上皮NCM460细胞的能量代谢具有损伤作用,其损伤作用与SIRT3相关。

AIM:To investigate the effect of deoxycholic acid(DCA)on the energy metabolism in human normal colon epithelial NCM460cells.METHODS:NCM460cells was treated with DCA at10,30and100μmol/L for5d,or DCA at100μmol/L for3,5and7d.After treated with DCA at100μmol/L for3d,the cells were treated with resveratrol,the activator of sirtuin3(SIRT3),for the next4d.Adenosine triphosphate(ATP)production in the mitochondria and lactate acid level were detected.The protein expression of SIRT3was determined by Western blot.RESULTS:DCA inhibited the ATP production,increased lactate acid level,and downregulated the protein expression of SIRT3in a dose-and time-dependent manner.Resveratrol at10μmol/L reversed the effects of DCA on the NCM460cells.CONCLUSION:DCA induces the dysfunction of energy metabolism in NCM460cells,and the mechanism may be related with SIRT3.