天津医药 ›› 2015, Vol. 43 ›› Issue (10): 1100-1103.

• 细胞与分子生物学 • 上一篇    下一篇

槲皮素对肝细胞氧化应激损伤的保护作用

  

  1. 1山东医学高等专科学校 (邮编250002); 2山东省医药生物技术研究中心
  • 收稿日期:2015-02-16 修回日期:2015-05-11 出版日期:2015-10-15 发布日期:2015-10-22

Protective effects of quercetin on hepatic cell damage induced by oxidative stress

  1. 1 Shandong Medical College, Jinan 250002, China2 Shandong Medicinal Biotechnology Centre
  • Received:2015-02-16 Revised:2015-05-11 Published:2015-10-15 Online:2015-10-22

摘要: 摘要: 目的 探讨槲皮素对氧化应激 Chang liver 细胞损伤的保护作用及其作用的分子机制。方法 培养
Chang liver 细胞, 将细胞随机分为正常对照组、 H2O2组及槲皮素低、 中、 高剂量组。MTT 法检测肝细胞的存活率; 流式
细胞术检测细胞凋亡率。2′,7′-二氯荧光素乙二酯(DCFH-DA)细胞内活性氧 (ROS) 荧光染色后, 荧光显微镜观察照
相及流式细胞仪检测细胞内活性氧水平。试剂盒测定细胞中超氧化物歧化酶(SOD)、 过氧化氢酶(CAT)、 谷胱甘肽
过氧化物酶 (GSH-Px) 活性及细胞中丙二醛 (MDA) 的含量。采用 Western blot 法检测 Nrf2 蛋白的表达。结果 H2O2
组较正常对照组细胞存活率、 SODGSH-PxCAT 活性降低 (P < 0.05), 凋亡率、 平均荧光强度(MFI)及 MDA 升高
(P < 0.05)。不同剂量槲皮素组细胞存活率及细胞内 SODGSH-PxCAT 活性高于 H2O2(P < 0.05)MDA、 凋亡率、
MFI 水平低于 H2O2(P < 0.05)。槲皮素低、 中及高剂量组 Nrf2 蛋白均高于 H2O2(P < 0.05)结论 槲皮素对氧化
应激肝细胞损伤具有保护作用, 其机制可能与激活 Nrf2-ARE 通路, 进而增强下游抗氧化基因的表达有关。

关键词: 槲皮素, 氧化性应激, 超氧化物歧化酶, 过氧化氢酶, 谷胱甘肽过氧化酶, Chang liver 细胞, Nrf2/ARE
通路

Abstract:

Abstract: Objective To explore the protective effects of quercetin on damage induced by oxidative stress and to clari⁃
fy its molecular mechanism. Methods Chang liver cell cultures were randomly divided into control groups, H2O2 group and
3 doses of quercetin groups. Cell survival rate was detected with MTT. Cell apoptotic rate was measured by FACSFluores⁃
cence-activated cell sorting. Intracellular reactive oxygen species (ROS) level in Chang liver cells were tested by flow cy⁃
tometer. The DCF fluorescence intensity of DCFH-DA-stained intracellular ROS was observed by fluorescence microscope.
The levels of malondialdehyde (MDA), superoxide dismutase(SOD), catalase (CAT) and glutathione peroxidase (GSH- Px)
were determined in liver cells using commercial available kits. The expression of Nrf2 were detected by Western blot. Re⁃
sults Compared with control, cell survival rate and levels of SOD, CAT and GSH-Px decreased significantly in H2O2 group
(P < 0.05 ),while cell appotosis rate, content of MDA and mean fluorescence intensity(MFI) increased in H2O2 group (P <
0.05). In comparison with H2O2, expression of Nrf2 protein was higher in all three quercetin treatment groups (P < 0.05). Con⁃
clusion Quercetin protected Chang liver cells from H2O2- induced oxidative stress, which may be caused by the increased ex⁃
pressions of down stream antioxidant genes via activating the Nrf2-ARE signaling pathway.

Key words: Quercetin, oxidative stress, superoxide dismutase, catalase, glutathione peroxidase, Chang liver cell, Nrf2/ARE pathway