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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2 h;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。
Objective: To investigate the effect of curcumin on Nrf2 nuclear translocation in rat Kupffer cells on the secretion of inflammatory cytokines induced by lipopolysaccharide (LPS). Methods: Kupffer cells were induced by 10μM, 20μM and 30μM respectively for 8h to induce nuclear translocation of Nrf2. Kupffer cells were randomly divided into control group, LPS group and intervention group. In control group, curcumin and LPS were used for normal control 10μg / mL LPS added Kupffer cell culture medium for 2 h; intervention group with 30μM curcumin intervention 8h, the rest with the LPS group. The level of nuclear translocation of Nrf2 was detected by Western blot, the levels of MDA and GSH were measured by spectrophotometry. The levels of TNF-α and IL-6 in supernatant were detected by ELISA, and IL-1β was detected by radioimmunoassay. RESULTS: (1) Curcumin induced nuclear translocation of Nrf2 in Kupffer cells, and nuclear translocation increased with increasing concentration. ② The MDA level in LPS group was significantly higher than that in control group (P <0.01). The MDA level in LPS group was significantly lower than that in LPS group (P <0.01), and still higher than that in control group (P <0.01). The level of GSH in LPS group was significantly lower than that in control group (P <0.01). The level of GSH in intervention group was significantly higher than that in LPS group (P <0.01), and still lower than that in control group (P <0.01). ③ The levels of TNF-α, IL-1β and IL-6 in supernatant of LPS group were significantly higher than those in control group (P <0.01), and were significantly lower in intervention group than those in model group (P <0.01) 0.01). CONCLUSION: Curcumin induces Nrf2 nuclear translocation in Kupffer cells, decreases the oxidative stress induced by LPS, and inhibits the secretion of inflammatory cytokines by Kupffer cells.