| 赵曙光李强王景杰王旭霞刘震雄闻勤生△.姜黄素诱导Nrf2 核转位对氧化应激诱导人肝细胞胰岛素抵抗的影响[J].现代生物医学进展英文版,2011,11(1):48-51. |
| 姜黄素诱导Nrf2 核转位对氧化应激诱导人肝细胞胰岛素抵抗的影响 |
| The Effect of Nrf2 Nuclear Translocation Induced by Curcumine onOxidative Stress-Mediated Insulin Resistance in Human Hepatocyte* |
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| DOI： |
| 中文关键词: Nrf2 姜黄素 肝细胞系L02 氧化应激 胰岛素抵抗 |
| 英文关键词: Curcumine Nrf2 L02 Oxidative stress Insulin resistance |
| 基金项目:陕西省中医药管理局中医药科研课题(2009jc53) |
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| 中文摘要: |
| 目的：研究姜黄素诱导转录因子NF-E2 相关因子2（NF-E2-related factor 2,Nrf2）核转位对氧化应激诱导人肝细胞L02 胰岛
素抵抗的影响。方法：用15μM 和30μM 姜黄素干预L02 肝细胞6 h 和l2 h，Western blot 检测Nrf2 核转位水平；将肝细胞分为
对照组、模型组、干预组，对照组用RPMI1640 正常培养，模型组用100U／L 葡萄糖氧化酶（GO）干预2 h，干预组用15 μM 和30
μM 姜黄素分别干预12 h 后给予100 U/L GO 干预2 h，各细胞均给予100 nM 胰岛素干预30 min。流式细胞术检测细胞内活性
氧簇(ROS)，用荧光强度（FI）来表示ROS 水平。分光光度法检测检测细胞MDA、GSH，葡萄糖氧化酶- 过氧化物酶法检测细胞培
养液中葡萄糖的水平，Western blot 检测胰岛素受体底物-1（IRS-1）磷酸化水平。结果：①姜黄素明显诱导Nrf2 核转位。②模型组
FI、MDA 水平较对照组显著升高(P<0.01)，干预组FI、MDA 水平均较模型组显著降低(P<0.01)，姜黄素15 μM 组FI、MDA 水平高
于30 μM 组(P<0.01)。模型组GSH 水平较对照组显著降低(P<0.01)，干预组GSH 水平较模型组显著升高(P<0.01)，姜黄素15
μM 组FI、MDA 水平高于30μM 组(P<0.01)。③模型组上清液葡萄糖浓度显著高于对照组（P<0.01），干预组上清液葡萄糖浓度
显著低于模型组（P<0.01），姜黄素15 μM 组上清液葡萄糖浓度高于30 μM 组(P<0.01)。模型组IRS-1 磷酸化水平较对照组降
低，干预组IRS-1 磷酸化水平均较模型组增高，姜黄素30 μM 组IRS-1 磷酸化水平高于15 μM 组。结论：姜黄素通过诱导Nrf2
核转位，降低细胞内氧化应激水平，进而逆转氧化应激诱导的胰岛素抵抗。 |
| 英文摘要: |
| Objective: To investigate the effect of nuclear factor E2-related factor 2 (Nrf2) translocation induced by curcumine on
oxidative stress mediated insulin resistance (IR) in human LO2 hepatocyte. Methods: The Nrf2 in nuclear fractions was assayed by western
blot analysis. The intercellular ROS level was analyzed on a flow cytometer by dihydroethidium and was indicated with fluorescence
intensity (FI). The levels of MDA and GSH were measured by spectrophotometric method. The glucose in culture medium was detected
by the glucose oxidizes peroxides method. The levels of IRS-1/p-IRS-1 were measured by western blot. Results: ①The levels of Nrf2 in
the nucleus increased markedly in hepatocytes treated by curcumine compared with that of the cells in control group. At the same time
point, nuclear Nrf2 was higher in cells treated by 30 μM curcumine than that in cells treated by 15 μM curcumine. Treated by the same
curcumine concentration, the nuclear Nrf2 level was higher in cells treated for 12 h than that in cells treated for 6 h. ②The levels of FI
and MDA increased significantly in model compared with that in the control group (P<0.01), both of which were significantly abrogated
in curcumine group (P<0.01). However, the levels of FI and MDA in 15 μM curcumine group were significantly higher than those in 15
μM curcumine group (P<0.01). Compared with control group, the level of GSH decreased significantly in model group(P<0,01), the
decreased GSH levels in model group were significantly abrogated in curcumine group (P<0.01), and the GSH levels in 15 μMcurcumine
group were significantly higher than that in 30 μM curcumine group (P<0.01). ③The glucose in culture medium increased significantly
in model group compared with that in control group （P<0.01）, the increased level was significantly abrogated in curcumine group (P<0.
01). However, the level in 15 μM curcumine group were significantly lower than that in 30 μM curcumine group (P<0.01). IRS-1
phosphorylation in model group decreased compared with that in control group and IRS-1 phosphorylation in curcumine group increased
in comparison with model group. The level of IRS-1 phosphorylation in 15 μM curcumine group was markedly lower than that in 30
μM curcumine group. Conclusion: Curcumine could induce Nrt2 nuclear translocation in cultured hepatocyte L02 and decrease intercellular
ROS, subsequently improve oxidative stress-mediated IR. |
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