论文部分内容阅读
目的探究二氢杨梅素(dihydromyricetin,DHM)对肝细胞脂质代谢的影响及其与SIRT1信号通路的关系。方法 0.2 mmol/L棕榈酸(palmitic acid,PA)处理HepG2细胞24 h,诱导为脂肪变性肝细胞模型,再用0、5、10、20μmol/L DHM及20μmol/L DHM+2μmol/L SIRT1抑制剂(EX527)分别干预24 h,CCK-8检测细胞增殖活性,油红0染色检测细胞脂滴形成情况,酶偶联比色法检测甘油三酯(triglyceride,TG)含量,蛋白免疫印迹法检测沉默信息调节因子1(silent information regulator 1,SIRT1)、腺苷酸活化蛋白激酶(adenosine monophosphate activated protein kinase,AMPK)、固醇调节元件结合蛋白1c(sterol regulatory element binding protein-1,SREBP-1c)、脂肪酸合成酶(fatty acid synthetase,FAS)、乙酰辅酶A羧化酶(acetyl-Co A carboxylase,ACC)的表达。结果 0.2 mmol/L PA和5、10、20μmol/L DHM对细胞增殖活力没有显著影响(P>0.05);5、10、20μmol/L DHM处理后明显减少脂肪变性的HepG2细胞脂滴蓄积和甘油三酯含量(P<0.01),增加磷酸化的AMPK(p-AMPK)和SIRT1的表达水平,并且降低脂质合成相关基因SREBP-1c、FAS、磷酸化的ACC(p-ACC)的蛋白表达,SIRT1抑制剂能显著增加脂肪变性的HepG2细胞的甘油三酯含量(P<0.01),削弱DHM对脂肪变性HepG2细胞SREBP-1c、FAS、P-ACC表达的抑制作用。结论 DHM通过上调脂肪变性HepG2细胞SIRT1信号通路改善肝细胞脂肪变性。
Objective To investigate the effect of dihydromyricetin (DHM) on lipid metabolism in hepatocytes and its relationship with SIRT1 signaling pathway. Methods HepG2 cells were treated with 0.2 mmol / L palmitic acid (PA) for 24 h, and induced by steatohepatitis model with 0, 5, 10 and 20 μmol / L DHM and 20 μmol / L DHM and 2 μmol / (EX527) for 24 h respectively. CCK-8 was used to detect the cell proliferation activity. The formation of lipid droplets was detected by oil red 0 staining. The content of triglyceride (TG) was detected by enzyme-linked immunosorbent assay (ELISA) Silent information regulator 1 (SIRT1), adenosine monophosphate activated protein kinase (AMPK), sterol regulatory element binding protein-1 (SREBP-1c) , Fatty acid synthetase (FAS) and acetyl-Co A carboxylase (ACC). Results No significant effect of 0.2 mmol / L PA and 5, 10, 20 μmol / L DHM on proliferation and proliferation of HepG2 cells was observed (P> 0.05). After 5, 10 and 20 μmol / L DHM treatment, lipid droplets and glycerol (P <0.01), increased the expression of phosphorylated AMPK (p-AMPK) and SIRT1, and decreased the protein expression of SREBP-1c, FAS and phosphorylated ACC (p-ACC) SIRT1 inhibitor could significantly increase triglyceride content (P <0.01) in steatosis HepG2 cells and attenuate the inhibitory effect of DHM on SREBP-1c, FAS and P-ACC expression in steatosis HepG2 cells. Conclusion DHM can improve steatosis of hepatocytes by up-regulating the SIRT1 signaling pathway in HepG2 cells.