银杏花化学成分对血管内皮细胞铁死亡的抑制作用  

The inhibitory effect of chemical components from Ginkgo biloba flower on ferroptosis in vascular endothelial cells

作　　者：李敏[1] 张丹 张林[1] 王立生 田瑛[1] 李彬 董俊兴 肖凤君 LI Min;ZHANG Dan;ZHANG Lin;WANG Li-sheng;TIAN Ying;LI Bin;DONG Jun-xing;XIAO Feng-jun(Institute of Radiation Medicine,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100850,China;Laboratory of Molecular Diagnosis and Regenerative Medicine,Medical Research Center,Affiliated Hospital of Qingdao University,Qingdao 266555,China)

机构地区：[1]军事科学院军事医学研究院辐射医学研究所,北京100850 [2]青岛大学附属医院医学研究中心分子诊断与再生医学实验室,青岛266555

出　　处：《国际药学研究杂志》2020年第10期857-862,共6页Journal of International Pharmaceutical Research

基　　金：国家“重大新药创制”科技重大专项资助项目(2018ZX09733-003);国家重点研发计划干细胞及转化研究专项(2017YFA0105303)。

摘　　要：目的探讨银杏花提取物及其中化学成分对人脐静脉内皮细胞(HUVEC)铁死亡的抑制作用,筛选出有效部位和有效化合物,为成药性提供初步研究数据。方法将银杏花70%乙醇提取物依次用不同有机溶剂萃取,得到4个萃取部位。利用erastin处理HUVEC细胞建立血管内皮细胞铁死亡模型,对4个萃取部位及前期从乙酸乙酯部位分离得到的9个化合物应用CCK-8法进行抗血管内皮细胞铁死亡活性筛选,并通过Western印迹法(WB)对其作用机制进行初步探讨。结果erastin诱导的HUVEC细胞死亡可被阳性药ferrostatin-1(Fer-1)逆转,是理想的血管内皮细胞铁死亡模型。银杏花总提物组能抑制eras-tin诱导的HUVEC细胞铁死亡,细胞存活率较erastin模型组明显提高(P<0.05),且乙酸乙酯部位组细胞存活率提高最多,为其活性部位。进一步对活性部位中分离获得的化合物进行活性筛选,结果表明,山柰酚、芹菜素、木犀草素及山柰酚-3-O-α-L-鼠李糖苷在10μmol/L时均能较模型组显著提高细胞存活率(P<0.01),其药效接近或超过Fer-1水平,其中木犀草素可使铁死亡细胞恢复至正常水平。WB结果显示,同模型组比,木犀草素可下调酰基辅酶A合成酶长链家族成员4和上调谷胱甘肽过氧化酶4的表达(P<0.01)。结论筛得银杏花乙酸乙酯部位为其抗血管内皮细胞铁死亡活性部位,木犀草素为其活性化合物,本研究将为抗血管炎症损伤药物研发提供新靶点和新的治疗策略。Objective To investigate the inhibitory effect of Ginkgo biloba flower extract and its chemical components on ferroptosis in human umbilical vascular endothelial cells(HUVEC),and to screen out the active fractions and compounds,so as to provide preliminary research data for drug development.Methods The 70%ethanol extract of G.biloba flower was extracted successively with different organic solvents to obtain four fractions.The ferroptosis of HUVEC cells was induced by treatment of the cells with erastin.The inhibitory activity on the ferroptosis was assayed for the four fractions and 9 compounds isolated from the ethyl acetate-extracted fraction by the CCK-8 method,and the related action mechanism was preliminarily investigated by Western blot(WB).Results The erastin-induced HUVEC cell death was reversed by the positive drug ferrostatin-1(Fer-1)and was thus an ideal model of ferroptosis in vascular endothelial cells.The total G.biloba flower extract inhibited the erastin-induced ferroptosis in HUVEC cells,as indicted by the cell survival rate that was significantly higer in the total G.biloba flower extract group than in the erastin model group(P<0.05),and the ethyl acetate-extracted fraction group showed the highest increase in the cell survival rate,demonstrating that this fraction was an active fraction from the total G.biloba flower extract.Furthermore,among the 9 compounds isolated from the active fraction,the flavonoids kaempferol,apigenin,luteolin and kaempferol-3-O-α-L-rhamnoside significantly icreased the cell survival rate,compared with the model group(P<0.01),and showed similar or even stronger effect than the Fer-1.Among them,luteolin restored the cell viability to a normal level.WB results showed that luteolin down-regulated the acyl-CoA synthetase long-chain family member 4 expression and up-regulated the glutathione peroxidase 4 expression,both significantly,compared with the model group(P<0.01).Conclusion The ethyl acetate-extracted fraction of the total G.biloba flower extract is the active fract

关 键 词：银杏花提取物 铁死亡 黄酮类化合物 

分 类 号：R96[医药卫生—药理学]