食品科学 ›› 2020, Vol. 41 ›› Issue (5): 108-114.doi: 10.7506/spkx1002-6630-20190301-010

• 营养卫生 • 上一篇    下一篇

榆干离褶伞溶栓酶对酒精损伤血管内皮细胞的保护作用

李芳芳,丛贺,沈明花   

  1. (延边大学医学院,吉林 延吉 133002)
  • 出版日期:2020-03-15 发布日期:2020-03-23
  • 基金资助:
    国家自然科学基金地区科学基金项目(81760031)

Protective Effect and Mechanism of Fibrinolytic Enzyme from Lyophyllum ulmarium on Alcohol-Induced Injury in Vascular Endothelial Cells

LI Fangfang, CONG He, SHEN Minghua   

  1. (Medical College, Yanbian University, Yanji 133002, China)
  • Online:2020-03-15 Published:2020-03-23

摘要: 目的:研究榆干离褶伞溶栓酶(fibrinolytic enzyme from Lyophyllum ulmarium,LUFE)对酒精所致人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)损伤的保护作用及其机制。方法:以酒精诱导HUVECs损伤,四甲基偶氮唑蓝法检测细胞活力,检测细胞培养上清液乳酸脱氢酶(lactate dehydrogenase,LDH)活力;比色法检测细胞超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活力和丙二醛(malondialdehyde,MDA)含量;吖啶橙-溴化乙啶双染法观察细胞凋亡;用荧光染料双氢罗丹明123和罗丹明123分别检测细胞总活性氧(reactive oxygen species,ROS)水平和线粒体跨膜电位;蛋白印迹法检测Bax、Bcl-2、细胞色素c、cleaved半胱氨酸天冬氨酸蛋白水解酶(cysteinyl aspartate specific proteinase,cleaved caspase)-9和cleaved caspase-3的表达水平。结果:LUFE能够提高内皮细胞存活率,降低LDH和MDA水平,提高SOD、GSH-Px活力,减少ROS的生成,并抑制酒精所致的线粒体跨膜电位的下降和细胞凋亡。Western blot结果表明,LUFE提高Bcl-2蛋白表达水平,并减少细胞色素c、cleaved caspase-9和cleaved caspase-3的表达。结论:LUFE对酒精所致血管内皮细胞的损伤有保护作用,其机制可能与其抗氧化、抑制酒精诱导的线粒体途径细胞凋亡有关。

关键词: 榆干离褶伞, 溶栓酶, 人脐静脉内皮细胞, 酒精, 保护作用

Abstract: Objective: To investigate the protective effect and mechanism of fibrinolytic enzyme from Lyophyllum ulmarium (LUFE) on alcohol-induced injury in human umbilical vein endothelial cells (HUVECs). Methods: Cell viability was evaluated by the methyl thiazolyl tetrazolium assay and the activity of lactate dehydrogenase (LDH) in cell-free culture supernatants was determined. The activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) and the content of malondialdehyde (MDA) were measured by colorimetry. The apoptosis changes of HUVECs were detected with acridine orange/ethidium bromide staining. The level of reactive oxygen species (ROS) and mitochondrial transmembrane potential (Δφm) were evaluated by dihydrorhodamine123 and rhodamine 123 staining, respectively. The protein expression of Bax, Bcl-2, cytochrome c, cleaved cysteinyl aspartate specific proteinase (caspase)-9 and cleaved caspase-3 was determined using Western blot assay. Results: Compared with the model group, LUFE could increase the survival rate, decrease the contents of LDH and MDA, and increase the activity of SOD and GSH-Px in HUVECs. LUFE could also reduce intracellular ROS production, increase mitochondrial transmembrane potential and inhibit apoptosis induced by alcohol. LUFE could down-regulate the expression of cytochrome c, cleaved caspase-9 and cleaved caspase-3, and up-regulate the expression of Bcl-2. Conclusion: LUFE has a protective effect on alcohol-induced injury of HUVECs, which may be related to its antioxidant capacity and its ability to regulate the mitochondrial signal pathway.

Key words: Lyophyllum ulmarium, fibrinolytic enzyme, human umbilical vein endothelial cells, alcohol, protective effect

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