中国动脉硬化杂志2024，Vol.32Issue(7)：558-566,9.DOI:10.20039/j.cnki.1007-3949.2024.07.002
氧化型脂蛋白(a)通过抑制细胞色素b表达促进血管内皮细胞焦亡
Oxidized lipoprotein(a)induces endothelial cell pyroptosis by inhibiting the expres-sion of cytochrome b
摘要
Abstract
Aim To explore the mechanism of oxidized lipoprotein(a)(oxLp(a))inducing pyroptosis of vascu-lar endothelial cells.Methods After incubating human umbilical vein endothelial cells(HUVEC)with 100 mg/L ox-Lp(a)for 24 hours,Western blot and RT-qPCR was used to detect pyroptosis related proteins,pro-inflammatory cytokines,mitochondrial related proteins NRF1,NRF2,PGC-1α and mitochondrial gene cytochrome b(CYTB),ELISA was used to detect the levels of inflammatory factors,scanning electron microscopy was used to detect cell membrane rup-ture,transmission electron microscopy was used to detect mitochondrial morphology,Hoechst33342/PI staining was used to detect cell apoptosis,MitoSOX probe was used to detect mitochondrial reactive oxygen species(mtROS),Flu-4AM probe was used to detect calcium ions,JC-1 probe was used to detect mitochondrial membrane potential(MMP),and Calcein AM staining was used to detect mitochondrial permeability transition pore(mPTP).Transfecting HUVEC with CYTB overexpressing lentivirus and analyzing its effects on oxLp(a)induced pyroptosis and mitochondrial function.Results After treatment with oxLp(a),the expression of NLRP3,pro-Caspase-1,Caspase-1,GSDMD and GSDMD-N proteins re-lated to pyroptosis were significantly increased(P＜0.05);the protein and mRNA levels of CYTB and pro-inflammatory cy-tokine IL-1β,IL-18 were significantly increased(P＜0.05).Small pores appeared on the cell membrane,the percentage of PI stained positive cells significantly increased(P＜0.05).OxLp(a)significantly inhibited the expression of mito-chondrial related proteins NRF1,NRF2 and PGC-1α,and the expression of mitochondrial gene CYTB,promoted an in-crease in mtROS generation,Ca2+overload,a decrease in ATP levels,a decrease in MMP,an increase in mPTP values,and abnormal mitochondrial morphology.After transfection with pHelper 2.0 lentivirus vector overexpressing CYTB,it was found that oxLp(a)induced HUVEC pyroptosis and mitochondrial morphological and functional abnormalities were par-tially reversed by overexpression of CYTB.Conclusion oxLp(a)promotes mitochondrial morphological and functional abnormalities and induces HUVEC pyroptosis by downregulating CYTB.关键词
细胞焦亡/氧化型脂蛋白(a)/线粒体功能/血管内皮细胞/细胞色素bKey words
cell pyroptosis/oxidized lipoprotein(a)/mitochondrial function/vascular endothelial cell/cytochrome b分类
医药卫生引用本文复制引用
曹子彤,陈彦君,谈世铭,饶羽祝,王晶晶,蔡泽民,王佐..氧化型脂蛋白(a)通过抑制细胞色素b表达促进血管内皮细胞焦亡[J].中国动脉硬化杂志,2024,32(7):558-566,9.基金项目
国家自然科学基金项目(81970389) （81970389）
湖南省科技创新计划项目(2021SK51801) （2021SK51801）
