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

摘要

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(10 μmol/L、30 μmol/L和100 μmol/L)作用5 d,或DCA(100 μmol/L)作用3、5和7 d;DCA(100 μmol/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 NCM460 cells.METHODS: NCM460 cells was treated with DCA at 10, 30 and 100 μmol/L for 5 d, or DCA at 100 μmol/L for 3, 5 and 7 d.After treated with DCA at 100 μmol/L for 3 d, the cells were treated with resveratrol, the activator of sirtuin 3 (SIRT3), for the next 4 d.Adenosine triphosphate (ATP) production in the mitochondria and lactate acid level were detected.The protein expression of SIRT3 was determined by Western blot.RESULTS: DCA inhibited the ATP production, increased lactate acid level, and downregulated the protein expression of SIRT3 in a dose-and time-dependent manner.Resveratrol at 10 μmol/L reversed the effects of DCA on the NCM460 cells.CONCLUSION: DCA induces the dysfunction of energy metabolism in NCM460 cells, and the mechanism may be related with SIRT3.