湖北科技学院学报（医学版）

目的 探讨多穗石柯总黄酮提取物(TC)对H_2O₂诱导人脐静脉内皮细胞(HUVECs)铁死亡的改善作用。方法 大鼠经TC给药后腹主动脉取血，制备含药大鼠血清，经CCK-8法确定其培养细胞的有效浓度，并验证其对H_2O₂诱导损伤的HUVECs的保护作用；Western blot检测各组细胞中CBSL、ACSL4的蛋白表达水平；提取细胞RNA进行转录组测序及生物信息学分析。结果 10%含药大鼠血清组的细胞活力值显著升高；H_2O₂处理后CBSL蛋白显著下调，而TC和Fer-1处理后细胞活力值显著升高；转录组测序显示，CBSL显著性下调，且与硫化合物代谢密切相关；Western blot结果显示：TC和Fer-1组效果相当，均能够逆转H_2O₂组中CBSL蛋白表达水平的下降以及ACSL4蛋白表达水平的升高。结论 TC能够减轻氧化应激介导的内皮细胞铁死亡，对血管具有潜在的保护作用。

Objective To explore the ameliorative effect of total flavonoid extract of Lithocarpus polystachyus Rehd(TC) on H_2O₂-induced ferroptosis in human umbilical vein endothelial cells(HUVECs).Methods Blood was taken from the abdominal aorta of rats after administration of TC to prepare drug-containing rat serum, and the effective concentration for cultured cells was detemined by CCK-8 method and its protective effect on H_2O₂-induced injury in HUVECs were evaluated.The protein expression level of Acyl-CoA synthetase long-chain 4(ACSL4) and Cystathionine β-synthase(CBSL) in cells of each group were detected by Western blot.The RNA of the cells was extracted to carry the transcriptome sequencing and bioinformatics analysis.Results Cell viability values were significantly higher in the 10% drug-containing rat serum group.CBSL protein was down-regulated in the presence of H_2O₂-induced damage and cell viability values were significantly higher after treatment with TC or Fer-1.Transcriptome sequencing results showed that CBSL was significantly down-regulated and closely related to sulfur compound metabolism.Additionally, Western blot results showed that TC and Fer-1 shared the similar effects to decrease the expression level of CBSL proteins and increase the expression level of ACSL4 proteins compared with H_2O₂ group.Conclusion TC attenuates oxidative stress-mediated ferroptosis in endothelial cells and shows the potential protective effects on blood vessels.