芹黄素对过氧化氢所致黑素细胞凋亡的作用

Effects of Apigenin on H_2O_2-induced Apoptosis in Melanocytes

目的采用过氧化氢(H2O2)诱导的黑素细胞体外氧化应激模型探讨芹黄素对黑素细胞的抗氧化保护作用。方法采用MTT法测定不同浓度芹黄素预处理前后H2O2对正常人表皮黑素细胞活力的影响,An-nexin-V/PI双染流式细胞术检测细胞的凋亡情况,DCFH-DA流式细胞术检测细胞活性氧(ROS)的含量。结果 250μmol/L的H2O2作用于黑素细胞24h,黑素细胞活力降低至(36.42±7.21)%,凋亡率增加至(48.82±12.55)%。0.6μmol/L,2.5μmol/L和10.0μmol/L芹黄素预处理1h后,H2O2所致细胞活力分别增加至(41.53±5.25)%,(45.33±6.28)%和(51.92±5.29)%,凋亡率减少至(42.25±7.69)%,(37.54±8.82)%和(32.25±58.28)%。250μmol/L的H2O2作用于黑素细胞2h后,黑素细胞ROS含量增加至空白对照组的(69.22±8.57)倍。10μmol/L芹黄素预处理1h后H2O2所致黑素细胞ROS含量为空白对照组的(54.48±5.03)倍,低于单用H2O2处理组。结论芹黄素可能通过减少ROS生成和抑制氧化应激对HO所致黑素细胞凋亡具有保护作用,其可能具有开发为治疗白癜风新药的前景。

Objective To investigate the antioxidant and protective effects of apigenin by using a model of H2O2-induced oxidative stress in melanocytes in vitro. Methods Effects of H2O2 on cell viability were measured by MTF assay. H2O2 -induced apoptosis in melanocytes were measured by annexin-V/PI staining cytometry analysis. H2O2 -triggered ROS generation was detected by cytometry analysis with DCFH-DA method. Results Treated with 250 μmol/L H2O2 for 24 h, the cell viability of melanocytes decreased to （ 36.42 ± 7.21 ） %, while the proportion of apoptotic cells increased to （48.82 ± 12.55 ）%. Pretreatment with 0.61μmol/L, 2.5μmol/L and 10μmol/L of apigenin significantly increased cell viability of H2O2 -treated malnocytes to （41.53 ± 5.25 ） %, （45.33 ± 6.28 ） % and （ 51.92 ± 5.29 ） % respectively, and decreased the proportion of apoptotic cells to （42.25 ± 7.69） %, （37.54 ± 8.82） % and （32.25 ± 58.28 ） % respectively. Treatment with H2O2 for 2h significantly increased ROS generation by （69.22 ±8.57） folds, while pretreatment with apigenin decreased the H2O2-indueed ROS generation to （54.48 ±5.03） folds. Conclusions Apigenin could protect melanocytes against H2O2-induced apoptosis probably by inhibiting the ROS generation and therefore the oxidative stress, thus may have a potential use in treating vitiligo.