荷叶碱对bel-7402细胞胆固醇代谢的影响

目的:探讨荷叶主要单体成分荷叶碱对Bel-7402细胞株胆固醇吸收、合成及酯化的影响。方法:体外培养Bel-7402细胞株,待细胞长满80%后无血清培养基饥饿细胞24h,分别加入1uM、10uM、100uM的荷叶碱及10uM的阿托伐他订,实时定量PCR和Western blotting法分别检测给药24小时后细胞内低密度脂蛋白受体(LDLR)、β-羟β-甲基戊二酰辅酶A还原酶(HMGCR)、酰基辅酶A胆固醇酰基转移酶(ACAT)的基因表达和蛋白质水平。结果:与对照组相比,荷叶碱各剂量组均能明显升高LDLR、HMGCOAR基因表达及蛋白水平,且呈剂量依赖性(P<0.01),阿托伐他汀组LDLR、HMGCOAR基因表达及蛋白水平高于荷叶碱各组(P<0.01),荷叶碱、阿托伐他汀均能降低细胞内ACAT基因表达及蛋白含量,荷叶碱中剂量组ACAT基因表达及蛋白含量均高于阿托伐他汀组(P<0.01,P<0.05),荷叶碱高剂量组ACAT基因表达高于阿托伐他汀组(P<0.05),蛋白水平两组没有显著性差异。结论:荷叶碱可能是通过抑制细胞内胆固醇的合成、抑制胆固醇酯酶的活性及升高低密度脂蛋白受体的量而起到降血脂作用。

Effect of nuciferine on cholesterol metabolism in Bel-7402 cells

Objective:To investigate effect of nuciferine, which is the main monomer of lotus leaf,on cholesterol metabolism in Bel-7402 cells.Methods:Bel-7402 cells were grown in 1640 medium with 10 % FBS for 24 h.The medium was replaced with serum-free medium and cells were incubated for another 24 h.1 mol/L, 10 mol/L and 100 mol/L nuciferine and 10 mol/L atorvastatin were added to the mediums respectively.Gene expression and protein level of LDLR, MHMGCoA and ACAT were measured by real time PCR and Western blotting.Results:LDLR,HMGCoA mRNA and protein levels increased significantly by each dose of nuciferine in a dose-dependent manner(P<0.01)when compared with the control group.The levels of LDLR, HMGCOAR mRNA and protein were higher in atorvastatin-loading cells than those in nuciferine-loading cells(P<0.01).The levels of ACAT mRNA expression and protein content were reduced by Nuciferine or Atorvastatin , higher in middle dose of nuciferine group than that in atorvastatin group(P<0.01, P<0.05).The levels of ACAT mRNA expression were higher in high dose of nuciferine group than those in atorvastatin group(P<0.05), while there was no significant difference in the level of protein between the two groups.Conclusion:Nuciferine is likely to inhibit the synthesis of cholesterol and cholesterol esterase activity, and elevate low-density lipoprotein receptor expression to reduce blood lipid.