对乙酰氨基酚通过过氧化物酶体增殖物激活受体γ共激活因子1α信号通路影响HepaRG细胞线粒体新生

发布时间：2018-05-24 21:54

本文选题：对乙酰氨基酚 + 线粒体　；参考：《中国药理学与毒理学杂志》2017年02期

【摘要】：目的探讨对乙酰氨基酚(APAP)对HepaRG细胞过氧化物酶体增殖物激活受体γ共激活因子1α(PGC-1α)信号通路介导的线粒体新生的影响。方法接种HepaRG细胞并给予生长培养基,待细胞长满后,替换为分化培养基进行诱导分化,每天观察细胞形态并拍照。APAP(0.125,0.25,0.5,1,2,4,8和12 mmol·L~(-1))处理诱导分化后的HepaRG细胞24和48 h,MTT法测定细胞存活率。Western蛋白印迹法检测细胞线粒体新生相关蛋白PGC-1α、核呼吸因子2(NRF-2)和线粒体转录因子A(TFAM),以及线粒体构成蛋白NADH脱氢酶亚基1(ND-1)的表达。结果诱导分化后显微镜下可见肝细胞样和胆管细胞样2种形态的细胞。与正常对照组相比,APAP作用24和48 h后,随APAP浓度的增加,细胞存活率不断降低,其IC_(50)分别5.64和2.65 mmol·L~(-1)。与正常对照组相比,APAP作用24 h,0.5,1,2和4 mmol·L~(-1)组PGC-1α表达水平显著增加(P0.01),8 mmol·L~(-1)组显著降低(P0.01);0.5 mmol·L~(-1)组NRF-2表达水平显著增加(P0.05),2,4和8 mmol·L~(-1)组显著降低(P0.01);1 mmol·L~(-1)组TFAM表达水平显著增加(P0.05),4和8 mmol·L~(-1)组显著降低(P0.01);0.5,1,2和4 mmol·L~(-1)组ND-1表达水平显著增加(P0.01),8 mmol·L~(-1)组显著降低(P0.01)。结论 APAP可诱导或抑制HepaRG细胞的线粒体新生,其机制可能与PGC-1α通路蛋白表达有关。
[Abstract]:Objective to investigate the effect of paracetamol (AP) on mitochondrial neogenesis mediated by peroxisome proliferator activated receptor 纬 -coactivator 1 伪 -PGC-1 伪 signal pathway in HepaRG cells. Methods HepaRG cells were inoculated and the growth medium was given. When the cells were full, the cells were replaced with the differentiation medium to induce differentiation. Observation of cell morphology and photo taking of .APAPN 0.125 ~ 0.25 ~ (0.25) and 12 mmol / L ~ (-1) treatment of differentiated HepaRG cells for 24 and 48 h MTT assay for cell viability. Western blot assay for the detection of mitochondrial neo-associated protein PGC-1 伪, nuclear respiratory factor 2NRF-2) and mitochondrial transcription. The expression of TFAM and NADH dehydrogenase subunit 1 ND-1. Results Hepatocyte-like cells and cholangiobiliary cell-like cells were observed under microscope after induced differentiation. Compared with the normal control group, the cell survival rate decreased with the increase of APAP concentration at 24 and 48 h after treatment with APAP, and its ICD 50 was 5.64 and 2.65 mmol / L ~ (-1), respectively. Compared with the normal control group, the expression of PGC-1 伪 increased significantly in the 24 h and 4 mmol groups (P 0.01 and 8 mmol / L).) the NRF-2 expression level in the P0.01 + 0. 05 mmol / L + 1) group was significantly lower than that in the control group. (2) the expression of NRF-2 in the P0. 055 and 8 mmol / L ~ (1) groups was significantly lower than that in the control group. (2) the expression level of TFAM in the P0. 01 + 1 mmol group was significantly higher than that in the control group. (0. 051 and 8 mmol / L ~-1) groups were significantly lower than those in the control group. The expression of ND-1 was significantly increased in the group of P0.01P0. 01 and 4 mmol. The expression of ND-1 was significantly increased in the group of P0. 01 and P0. 01 mmol. (1) the expression level of P0. 01 was significantly decreased in the group of P0. 01 and P0. 01 of P0. 01. Conclusion APAP can induce or inhibit mitochondrial neogenesis in HepaRG cells, and its mechanism may be related to the expression of PGC-1 伪 pathway protein.
【作者单位】：军事医学科学院疾病预防控制所毒理学评价研究中心;
【基金】：国家自然科学基金青年科学基金(81302864) 联合利华国际合作项目(CH-2011-1318)~~
【分类号】：R96

【相似文献】