柴胡皂苷A通过抑制氧化应激和铁死亡减轻过氧化氢诱导的人脐静脉内皮细胞损伤

Saikosaponin A inhibits oxidative stress and ferroptosis and reduces the injury of human umbilical vein endothelial cells induced by hydrogen peroxide

目的探讨柴胡皂苷A对过氧化氢(H_(2)O_(2))诱导的人脐静脉内皮细胞(HUVEC)氧化应激和铁死亡的影响。方法利用H_(2)O_(2)诱导建立HUVEC损伤模型,设置对照组、H_(2)O_(2)组及柴胡皂苷A低、中、高剂量组。用CCK-8法检测细胞的存活率;试剂盒法检测细胞内丙二醛(MDA)、谷胱甘肽(GSH)水平和超氧化物歧化酶(SOD)活性;qRT-PCR法测定谷胱甘肽过氧化物酶4(GPX4)、长链酯酰辅酶A合成酶4(ACSL4)的mRNA含量;Western blot法检测GPX4、ACSL4的蛋白表达。结果与对照组相比,H_(2)O_(2)组HUVEC存活率显著下降(P<0.05),GSH水平、SOD活性显著降低(P<0.05),MDA水平显著升高(P<0.05),GPX4 mRNA及蛋白表达显著降低(P<0.05),ACSL4 mRNA及蛋白表达显著升高(P<0.05);与H_(2)O_(2)组相比,柴胡皂苷A能呈剂量依赖性地提高HUVEC存活率,抑制氧化应激,提高GSH水平和SOD活性(P<0.05),降低MDA水平(P<0.05);此外,柴胡皂苷A呈浓度依赖性抑制铁死亡,上调GPX4 mRNA及蛋白的表达(P<0.05),降低ACSL4 mRNA及蛋白的表达(P<0.05)。结论柴胡皂苷A对H_(2)O_(2)诱导的HUVEC氧化应激和铁死亡有拮抗作用。

Aim To investigate the effect of saikosaponin A on oxidative stress and ferroptosis of human umbilical vein endothelial cells(HUVEC)induced by hydrogen peroxide(H_(2)O_(2)).Methods The oxidative injury model in HUVEC was established and cells were divided into control group,model group,saikosaponin A low-dose,medium-dose and high-dose groups.CCK-8 assay was used to detect cell viability.The levels of malondialdehyde(MDA),glutathione(GSH)and superoxide dismutase(SOD)were detected by the kit assay.qRT-PCR was used to detect the mRNA levels of glutathione peroxidase 4(GPX4)and acyl-CoA synthetase long chain family member 4(ACSL4).The protein expression of GPX4 and ACSL4 were detected by Western blot.Results Compared with the control group,the survival rate of HUVEC,GSH level and SOD activity in H_(2)O_(2) group were significantly decreased(P<0.05),the level of MDA was significantly increased(P<0.05),the expressions of GPX4 mRNA and protein were significantly decreased(P<0.05),the expressions of ACSL4 mRNA and protein were increased(P<0.05).Compared with the model group,saikosaponin A dose-dependently increased the survival rate of HUVEC,inhibited oxidative stress,improved the activity of SOD and GSH level(P<0.05),and decreased the level of MDA(P<0.05);In addition,saikosaponin A inhibited ferroptosis in a concentration-dependent manner,up-regulated the expression of GPX4 mRNA and protein(P<0.05),and decreased the expression of ACSL4 mRNA and protein(P<0.05).Conclusion Saikosaponin A can protect HUVEC from oxidative stress and ferroptosis induced by H_(2)O_(2).