槲皮素对小鼠ADTC-5细胞影响的体外试验△

Effects of quercetin on mouse ADTC-5 cells in vitro

摘要:

[目的]探讨槲皮素（quercetin, Quer）调控核因子 E2 相关因子（nuclear factor erythroid 2 related factor, NRF2）-血红素氧合酶 1 （heme oxygenase1, HO1） 信号通路对小鼠 ADTC-5 细胞的作用机制。[方法] 体外培养小鼠 ADTC-5 细胞，分成 3 组。Contr 组，不予特殊处理；IL-1β 组，给予 10 ng/mL IL-1β 处理 24 h；IL-1β+Quer 组，经 10 ng/mL IL-1β 处理 24 h 后给予 100 μmol/L Quer 处理 24 h。行比色法、Western blot、qRT-PCR 检测二价铁离子（Ferrousion, Fe2+ ）等的含量、蛋白及 mRNA 表达水平。[结果] 比色法检测显示，与 Contr 组比较，IL-1β 组和 IL-1β+Quer 组的 Fe2+ [(6.6±0.2) μmol/L vs (13.9±0.2) μmol/L vs (9.2±0.2) μmol/L, P＜0.001] 、丙二醛 （malondialdehyde, MDA） [(2.2±0.1) nmol/mL vs (6.1±0.0) nmol/mL vs (3.6±0.1) nmol/mL, P＜ 0.001] 含量显著增加，超氧化物歧化酶 （superoxide dismutase, SOD） [(58.0±0.7) U/mL vs (26.1±0.8) U/mL vs (47.8±1.0) U/mL, P＜ 0.001] 活力显著下降。Western blot 检测和 qRT-PCR 检测显示，小鼠 ADTC-5 细胞上清谷胱甘肽过氧化物酶4（glutathione peroxi- dase 4, GPX4）、NRF2、HO1 蛋白表达水平和 mRNA 表达水平均为 Contr 组＞IL-1β+Quer 组＞IL-1β 组；小鼠 ADTC-5 细胞上清 IL-1β、MMP-13 蛋白表达水平和 mRNA 表达水平均为 IL-1β 组＞IL-1β+Quer 组＞Contr 组。[结论]槲皮素上调 NRF2-HO1 信号通路抑制小鼠 ADTC-5 细胞铁死亡。

Abstract:

[Objective] To explore the role of quercetin (Quer) regulating nuclear factor erythroid 2 related factor (NRF2) by heme oxygenase1 (HO1) signaling pathway on mouse ADTC- 5 cells. [Methods] Mouse ADTC- 5 cells were cultured in vitro and divided into 3 groups. Control group (Contr group) had no special treatment, while the IL-1β group was treated with 10 ng/ MLL-1β for 24 hours, and the IL-1β+Quer group was treated with 10 ng/mL IL-1β for 24 hours and then treated with 100 μmol/L Quer for 24 hours. Colorimetry, western blot and qRT-PCR assays were performed to detect ferrousion (Fe2+ ), protein and mRNA expressions of the factors. [Results] In term of colorimetric detection, compared with the contr group, the IL-1β and IL-1β+Quer groups measured Fe2+ [(6.6±0.2) μmol/L vs (13.9±0.2) μmol/L vs (9.2±0.2) μmol/L, P＜0.001], malondialdehyde (MDA) [(2.2±0.1) nmol/mL vs (6.1±0.0) nmol/mL vs (3.6±0.1) nmol/mL, P＜0.001] was significantly increased. However, compared with the contr group, the latter two groups measured superoxide dismutase (SOD) significantly decreased [(58.0±0.7) U/mL vs (26.1±0.8) U/mL vs (47.8±1.0) U/mL, P＜0.001]. As results of western blot and qRT-PCR assays, the protein and mRNA expression levels of glutathione peroxidase 4 (GPX4), NRF2 and HO1 in mouse ADTC-5 cells were ranked up-down as the contr group ＞the IL-1β+Quer group＞the IL-1β group, whereas the protein and mRNA expressions in the supernatant IL-1β and MMP-13 in mouse ADTC-5 cells were all ranked as the IL-1β group ＞the IL-1β+Quer group ＞the contr group. [Conclusion] The quercetin up-regulates NRF2-HO1 signaling pathway to inhibit iron death in mouse ADTC-5 cells.