DEHP经sirt1/pgc-1a通路诱导的HepG2细胞线粒体损伤效应

卜玲玲; 刘焕; 杨娇; 黄卓权; 靳登鹏; 柯倩华; 柳春红

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中国环境科学 ›› 2020, Vol. 40 ›› Issue (8) : 3621-3626.

环境毒理

DEHP经sirt1/pgc-1a通路诱导的HepG2细胞线粒体损伤效应

卜玲玲, 刘焕, 杨娇, 黄卓权, 靳登鹏, 柯倩华, 柳春红

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DEHP induced mitochondrial damage through sirt1/pgc-1a pathway in HepG2 cells

BU Ling-ling, LIU Huan, YANG Jiao, HUANG Zhuo-quan, JIN Deng-peng, KE Qian-hua, LIU Chun-hong

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摘要

为研究邻苯二甲酸二（2-乙基己基）酯（DEHP）是否通过sirt1/pgc-1a通路对细胞产生损伤效应，通过体外培养HepG2细胞，在1.6， 8， 40， 200， 1000mmol/L DEHP处理24或48h后，采用CCK-8测定细胞活力，ATP试剂盒检测细胞内ATP含量，NO试剂盒检测细胞上清NO含量，ELISA试剂盒检测细胞上清炎症因子TNF-a、IL-6的含量；同时在DEHP作用于HepG2细胞24或48h后，用Western blot检测线粒体调控基因sirt1、pgc-1a，nrf1、tfam的蛋白表达.结果表明：不同浓度的DEHP处理24， 48h后，细胞活力在高剂量都呈显著下降趋势.DEHP能引起ATP含量的显著下降、炎症因子含量的显著提高，但NO含量无显著变化.4种线粒体调控基因的蛋白表达水平sirt1、pgc-1a，nrf1、tfam在24h时无显著变化.而在48h时，随着剂量增加蛋白含量呈上升后下降趋势，并且sirt1在8mmol/L呈显著上升趋势，随后每个蛋白都在1000mmol/L显著下降（P < 0.05）.DEHP能引起HepG2细胞氧化应激，并通过影响sirt1/pgc-1a信号通路表达来影响线粒体生物合成从而造成细胞损伤.

Abstract

In order to explore whether DEHP could damage cells through sirt1/pgc-1a pathway, HepG2 cells were cultured with different concentrations of DEHP at 1.6,8,40, 200,1000mmol/L for 24 or 48 hours. Cell viability was measured by CCK-8 assay. The intracellular ATP content and NO in the supernatants of cell cultures were measured by corresponding test kits. The level of TNF-a and IL-6 in cell supernatants were measured by enzyme-linked immunosorbent assay (ELISA), and the protein expression of sirt1, pgc-1a, nrf1, tfam were detected by Western blot. The results showed that: After treated with different concentrations of DEHP for 24 and 48 hours, the cell viability decreased significantly at high doses. DEHP could cause the decrease of ATP content and the increase of inflammatory factor content. No significant changes for the protein expression levels of sirt1, pgc-1a, nrf1, tfam had been found when exposed for 24 h. When exposed for 48 h, the protein content increased first and then decreased with the increase of dose, and sirt1 increased significantly at 8mmol/L. At the high dose of 1000mmol/L, all proteins decreased significantly. This study suggested that DEHP could induce the expression of sirt1/pgc-1a signal pathway to affect the mitochondrial biosynthesis and cause cell damage.

导出引用

卜玲玲, 刘焕, 杨娇, 黄卓权, 靳登鹏, 柯倩华, 柳春红. DEHP经sirt1/pgc-1a通路诱导的HepG2细胞线粒体损伤效应[J]. 中国环境科学. 2020, 40(8): 3621-3626

BU Ling-ling, LIU Huan, YANG Jiao, HUANG Zhuo-quan, JIN Deng-peng, KE Qian-hua, LIU Chun-hong. DEHP induced mitochondrial damage through sirt1/pgc-1a pathway in HepG2 cells[J]. China Environmental Science. 2020, 40(8): 3621-3626

中图分类号： X503.1

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