摘要
目的研究阿托伐他汀在炎症状态下对人肝癌细胞株Hep G2细胞氧化应激和脂质积聚的影响。方法将普通Hep G2细胞分为为3组:正常组、模型组[用100 ng·m L^(-1)肿瘤坏死因子(TNF-α)处理]及实验组(用100 ng·m L^(-1)TNF-α+10μmol·L^(-1)阿托伐他汀处理),3组同时负荷100μg·m L^(-1)低密度脂蛋白(LDL),处理时间24 h。用油红O染色测定细胞内脂质含量;以荧光定量聚合酶链式反应和免疫印迹法分别检测Hep G2细胞脂肪酸合酶(FAS)、乙酰辅酶A羧化酶(ACC)和固醇调节元件结合蛋白1(SREBP1)的基因和SREBP1蛋白表达;以二氯二氢荧光素-乙酰乙酸酯(DCFH-DA)荧光探针染色检测Hep G2细胞内活性氧化产物(ROS)的含量;以比色法检测Hep G2细胞过氧化氢(H2O2)和丙二醛(MDA)的含量。结果正常组与模型组SREBP1蛋白灰度值分别为1.01±0.001,1.61±0.34,这2组的SREBP1 mRNA水平分别为1.01±0.16,3.61±0.39,这2组的FAS的mRNA水平分别是1.03±0.32,1.99±0.36,这2组的ACC的mRNA水平分别是0.95±0.29,2.37±0.52。与正常组相比,模型组的细胞FAS、ACC、SREBP1的基因和SREBP1的蛋白表达明显增加,差异均有统计学意义(均P<0.05)。与模型组相比,实验组的Hep G2FAS、ACC、SREBP1的基因和SREBP1的蛋白灰度值分别是2.95±0.92,3.99±1.16,2.85±0.91,2.94±0.65,实验组的Hep G2 FAS、ACC、SREBP1的基因和SREBP1的蛋白表达明显增加,差异均有统计学意义(均P<0.05)。表明在炎症状态下,阿托伐他汀进一步加重了Hep G2细胞内脂质的沉积。正常组、模型组与实验组的细胞ROS含量(以荧光强度代表)分别为1.00±0.20,1.77±0.25,3.20±0.53;正常组、模型组与实验组的H2O2含量分别为(2.30±0.31),(4.32±0.77),(5.31±0.75)nmol·mg^(-1);这2组的MDA含量分别为(0.78±0.22),(1.86±0.23),(3.43±1.15)nmol·mg^(-1),与正常组比较,模型组差异均有统计学意义(均P<0.05);与模型组比较,实验组差异均有统计学意义(均P<0.05)。结论在炎症状态下,�
Objective To investigate the effect of atorvastatin on oxida- tive stress and intracellular lipid accumulation in HepG2 cells under in- flammatory state and explore the underlying mechanism. Methods HepG2 cells were treated with 100 ng · mL^-1 TNF- el, 100 ng · mL^-1 TNF - α ＋ 10 μmol · L^-1 atorvastatin in the presence of LDL for 24 h. Oil red 0 staining was used to examine the intracellular lipid contents.The mRNA and protein expressions of lipogenic genes （ FAS, ACC and SREBP1 ） were detected by real - time poly- merase chain reaction and Western blot. ROS levels were measured with the fluorescent probe of DCFH - DA. Contents of H202 and MDA were determined using the colorimetric method. Results Compared with normal group（the gray value of SREBP1 was 1.01 ±0. 001 ）, the gray value of SREBP1 in model group was 1.61 ±0. 34. The mRNA levels in normal group of SREBP1, FAS, ACC respectively were 1.01 ± 0. 16,1.03 ±0. 32,0. 95 ±0. 29, the values in model group respectively were 3.61 ±0. 39, 1.99 ±0. 36, 2. 37± 0. 52, the differences were statistically significantly （P 〈 0. 05 ）. Compared with model group, the mRNA levels of SREBP1, FAS, ACC and the gray value of SREBP1 in experimental group respectively were 2. 95±0. 92,3.99 ±1.16,2. 85± 0. 91,2. 94±0. 65, the differences were statisti- cally significantly（ P 〈0. 05 ）. At the same time, compared with normal group, the levels of ROS（ fluorescenceintensi- ty） ,H202 ,MDA respectively were 1.00 ＋0. 20,and （2. 30 ±0. 31 ） （0. 78 ＋0. 22） nmol ~ mg-~, the levels in model group respectively were 1.77 ±0. 25 and （4. 32 ±0. 77 ）, （ 1.86 ±0. 23 ） nmol · mg^-1, the differences were statistically significantly （ P 〈 0. 05 ）. Compared with model group, the levels of ROS, H2 O2, MDA in HepG2 cells in experimental group respectively were 3.2 ±0. 53 and （5.31 ±0. 75）, （3.43 ±1.15） nmol · mg^-1 ,the differences were statistically significantly （ P 〈 0. 05 ）. Conclusion Atorvastatin induced int
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2017年第9期802-805,共4页
The Chinese Journal of Clinical Pharmacology
基金
国家自然科学基金资助项目(31540029)
重庆市第七批市科技计划基金资助项目(cstc2016jcyj A0545)
