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

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。

Role of SIRT3 in dysfunction of energy metabolism induced by deoxycho-lic acid in human colon NCM460 cells

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.