姜黄素诱导Nrf2核转位对脂多糖引起库普弗细胞分泌炎症细胞因子的影响

目的：研究姜黄素诱导大鼠Kupffer细胞Nrf2核转位对脂多糖（LPS）引起的炎症细胞因子分泌的影响。方法：分别用10μM、20μM和30μM干预Kupffer细胞8h，诱导Nrf2核转位水平；将Kupffer细胞随机分为对照组、LPS组和干预组，对照组正常培养未加姜黄素和LPS，LPS组用10μg／mL的LPS加入Kupffer细胞培养液共同培养2h；干预组用30μM姜黄素干预8h后，余处理同LPS组。Western blot检测Nrf2核转位水平，分光光度法检测细胞MDA、GSH水平，ELISA法检测上清液TNF-α和IL-6，放免法检测IL-1β。结果：①姜黄素诱导Kupffer细胞Nrf2核转位，核转位水平随浓度增加而增高。②LPS组MDA水平较对照组显著升高（P〈0．01），干预组MDA水平较LPS组显著降低（P〈0．01），仍显著高于对照组（P〈0．01）。LPS组GSH水平较对照组显著降低（P〈0．01），干预组GSH水平较LPS组显著升高（P〈0．01），仍显著低于对照组（P〈0．01）。③LPS组上清液TNF-α，IL-1β和IL-6显著高于对照组（P〈0．01），干预组均显著低于模型组（P〈0．01），但显著高于对照组（P〈0．01）。结论：姜黄素通过诱导Kupffer细胞Nrf2核转位，降低LPS诱导的氧化应激损伤，抑制Kupffer细胞分泌炎症细胞因子。

The Effect of Nrf2 Nuclear Translocation Induced by Curcumine on Lipopolysaccharide Mediated Inflammatory Cytokine Expression in Rat Kupffer Cells

Objective： To investigate the effect of nuclear factor E2-related factor 2 （Nrf2） translocation induced by curcumine on lipopolysaecharide （LPS） mediated inflammatory cytokine expression in rat Kupffer cells. Methods： Kupffer cells were incubated in culture with 10 μM, 20 μM and 30 μM curcumine for 8 h to induce Nrf2 nuclear transloeation. Kupffer cell was divided into eontrol group, LPS and curcumine group. Cells in control group were incubated in culture only with RPMI 1640, cells in LPS group were treated with 10 μg/mL LPS for 2 h, cells in curcumine group were treated with 30 μ M curcumine for 8 h and subsequently treated with 10 12 g/mL LPS for 2 h. The Nrf2 in nuclear and cytoslic fraction was assayed by western blot analysis. The levels of MDA and GSH were measured by spectrophotometric method. The levels of TNF-a and IL-6 in culture medium were detected by the ELISA method, and the level of IL-1β was measured by radioimmunoassay. Results： ①The levels of Nrf2 in the nucleus increased markedly in Kupffer cells treated by different concentrations of curcumine compared with that of the cells in control group, and the level of nuclear Nrf2 increased with the concentration of curcumine. ②The level of MDA increased significantly in LPS group compared with that in the eontrol group （P〈0.01）, which was significantly abrogated in curcumine group （P〈0.01）. Compared with that of the control group, the level of GSH decreased signifieantly in model group（P〈0,01）, the decreased GSH level in LPS group significantly abrogated in eurcumine group （P〈0.01）. ③The levels of TNF-α , IL-1β and IL-6 in culture medium increased significantly in LPS group compared with that in control group （P〈0.01）, the increased level was significantly abrogated in cureumine group （P〈0.01）. Conclusion： Curcumine can induce Nrf2 nuclear transloea- tion in cultured Kupffer cells and decrease intercellular ROS, subsequently inhibit LPS-mediated inflammatory cytokine expression.