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目的:探索花青素对H2O2引起的Huh7细胞氧化应激、细胞凋亡的作用及其机制。方法:对H2O2和花青素干预培养的Huh7细胞,应用MTT法检测细胞活力,荧光探针DCFH-DA测定细胞内活性氧(ROS)生成量,免疫印迹测定Akt、磷酸化激活的c-Jun蛋白水平。结果:H2O20.8mmol/L孵育1小时可显著诱导Huh7细胞损伤,细胞活力下降到(49.27±3.2)%,ROS生成量是未处理细胞的3.56倍。细胞经花青素50μmol/L与H2O2共孵育后,细胞存活率提高到(81.2±2.34)%;花青素能显著抑制H2O2引起的Huh7细胞ROS生成,ROS生成下降74%(P<0.01)。花青素抑制H2O2引起Huh7细胞死亡和ROS生成的效应随剂量增加而加强。花青素抑制H2O2激发Huh7细胞磷酸化c-Jun表达,提高细胞Akt水平。结论:花青素抑制H2O2引起的Huh7细胞氧化应激损伤所导致的细胞死亡,其作用机制在于减少细胞内活性氧生成,抑制H2O2激活磷酸化c-Jun,提高细胞Akt水平。
Objective: To explore the effect of anthocyanin on oxidative stress and apoptosis of Huh7 cells induced by H2O2 and its mechanism. Methods: The viability of Huh7 cells cultured with H2O2 and anthocyanin was detected by MTT assay. The amount of intracellular reactive oxygen species (ROS) was detected by fluorescent probe DCFH-DA. Akt, phosphorylated c-Jun Protein level. Results: After being incubated at 20.8mmol / L for 1 hour, Huh7 cells were significantly injured and the viability of Huh7 cells decreased to (49.27 ± 3.2)%. The amount of ROS produced was 3.56 times of that of untreated cells. Cell viability increased to (81.2 ± 2.34)% after incubated with 50μmol / L anthocyanin and H2O2. Anthocyanin significantly inhibited the ROS production induced by H2O2 in Huh7 cells, and the ROS production decreased by 74% (P <0.01) . The effect of anthocyanin on H2O2-induced Huh7 cell death and ROS production increased with dose. Anthocyanin inhibited H2O2-induced phosphorylation of Huh7 cells c-Jun expression and increased cell Akt levels. CONCLUSION: Anthocyanin inhibits H2O2-induced cell death induced by oxidative stress in Huh7 cells. The mechanism of action is to reduce intracellular reactive oxygen species (ROS) production, inhibit H2O2-induced phosphorylation of c-Jun, and increase Akt level.