炎症状态下阿托伐他汀对HepG2细胞氧化应激和脂质积聚的影响

发布时间：2018-03-07 23:06

本文选题：阿托伐他汀　切入点：炎症应激　出处：《中国临床药理学杂志》2017年09期 　论文类型：期刊论文

【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P0.05)。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P0.05)。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P0.05);与模型组比较,实验组差异均有统计学意义(均P0.05)。结论在炎症状态下,阿托伐他汀诱导Hep G2细胞氧化应激,激活SREBP1/FAS/ACC通路,加重细胞内脂质积聚。
[Abstract]:Objective to study the effects of Atto vastatin on oxidative stress and lipid accumulation in Hep G2 cells under inflammatory condition. Model group [100ng 路mL ~ (-1)) TNF- 伪] and experimental group (100ng 路mL ~ (-1) TNF- 伪 10 渭 mol 路L ~ (-1)) Atto vastatin were treated with 100 渭 g 路m ~ (-1) mol 路L ~ (-1) low density lipoprotein (LDL). Fluorescence quantitative polymerase chain reaction (FQ-PCR) and Western blot were used to detect the gene and SREBP1 expression of fatty acid synthase (FASN), acetyl coenzyme A carboxylase (Hep) and steroid regulatory element binding protein (SSREBP1) in Hep G2 cells. The content of reactive oxidation product (Ros) in Hep G2 cells was detected by DCFH-DAA fluorescence probe staining. Results the grayscale values of SREBP1 protein in normal group and model group were 1.01 卤0.001 卤0.34 and 1.01 卤0.16 卤3.61 卤0.39, respectively. The mRNA level of FAS in these two groups was 1.03 卤0.32 卤0.36, respectively. The mRNA levels of ACC were 0.95 卤0.29 and 2.37 卤0.52, respectively. In the model group, the gene expression of FASA ACC-SREBP1 and the protein expression of SREBP1 were significantly increased (all P 0.05), compared with the model group, the expression of FASACC-SREBP1 was significantly higher than that of the model group (P < 0.05). The grayscale values of Hep G2FASA ACC-SREBP1 gene and SREBP1 protein were 2.95 卤0.92 卤1.16 ~ 3.99 卤0.91 卤2.94 卤0.65, respectively. The expression of Hep G2FASA ACC-SREBP1 gene and SREBP1 protein in the experimental group were significantly increased (all P0.05), indicating that the expression of Hep G2FAS-ACC-SREBP1 protein was significantly higher in the experimental group than that in the control group (P0.05). Atto vastatin further aggravated lipid deposition in Hep G2 cells. The ROS content in normal group, model group and experimental group (represented by fluorescence intensity) was 1.00 卤0.20 卤0.253.20 卤0.53, respectively. The content of H _ 2O _ 2 in the model group and the experimental group was 2.30 卤0.31 卤0.77 nmol 路mg ~ (-1), respectively, and the MDA content in the two groups was 0.78 卤0.22 nmol 路mg ~ (-1) and 1.86 卤0.23 ~ (1) nmol 路mg ~ (-1), respectively. There were significant differences between the model group and the normal group (P 0.05). Conclusion under the condition of inflammation, Atto vastatin induces oxidative stress in Hep G2 cells, activates SREBP1/FAS/ACC pathway and exacerbates intracellular lipid accumulation.
【作者单位】：重庆医科大学脂糖代谢性疾病重庆市重点实验室;
【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)
【分类号】：R96

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