胡黄连苷Ⅱ对ox-LDL诱导血管内皮细胞损伤影响  

Effects of picrosideⅡon ox-LDL-induced vascular endothelial cell injury

作　　者：吕静静 LU Jingjing(Zhengzhou Hospital of Traditional Chinese Medicine,Zhengzhou 450000,China)

机构地区：[1]郑州市中医院,河南郑州450000

出　　处：《青岛大学学报(医学版)》2025年第1期73-78,共6页Journal of Qingdao University(Medical Sciences)

基　　金：河南省中医药科学研究专项课题(2018ZY3-008、2023ZY2151)。

摘　　要：目的探讨胡黄连苷Ⅱ(P-Ⅱ)对氧化低密度脂蛋白(ox-LDL)诱导的血管内皮细胞损伤的影响及其机制。方法将EVC-304细胞分为对照组,ox-LDL组,P-Ⅱ低、中、高剂量组,mimic-NC组,miR-101-3p mimic组。采用双荧光素酶报告基因实验验证miR-101-3p和基质金属蛋白酶15(MMP15)的关系;实时荧光定量PCR(qRT-PCR)检测细胞中miR-101-3p表达;MTT法、划痕实验、Transwell小室和流式细胞仪分别检测细胞增殖、迁移、侵袭和凋亡;ELISA试剂盒检测细胞中白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、乳酸脱氢酶(LDH)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)水平;蛋白免疫印迹法(Western blot)检测细胞中增殖细胞核抗原(PCNA)、B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)及MMP15蛋白的表达水平。结果MMP15为miR-101-3p靶基因。与对照组比较,ox-LDL组miR-101-3p、IL-6、IL-1β、TNF-α、MDA、Bax表达水平以及细胞凋亡率、LDH释放率均显著升高(F=100.7~526.1,P<0.05),细胞增殖能力、划痕愈合率、细胞侵袭数目、GSH-Px活性以及PCNA、Bcl-2、MMP15蛋白表达水平显著降低(F=93.0~260.6,P<0.05)。与ox-LDL组比较,P-Ⅱ低、中、高剂量组miR-101-3p、IL-6、IL-1β、TNF-α、MDA、Bax表达水平以及细胞凋亡率、LDH释放率均显著降低(P<0.05),细胞增殖能力、划痕愈合率、细胞侵袭数目、GSH-Px活性以及PCNA、Bcl-2、MMP15蛋白表达水平显著升高(P<0.05)。上调miR-101-3p的表达可减弱P-Ⅱ对ox-LDL诱导的EVC-304细胞增殖、迁移、侵袭的促进作用以及对凋亡、氧化应激和炎症反应的抑制作用(P<0.05)。结论P-Ⅱ可能通过调控miR-101-3p/MMP15轴减轻ox-LDL诱导的EVC-304细胞损伤。Objective To investigate the effects of picrosideⅡ(P-Ⅱ)on vascular endothelial cell injury induced by oxidized low density lipoprotein(ox-LDL)and its mechanism of action.Methods EVC-304 cells were divided into control group,ox-LDL group,low-,medium-,and high-dose P-Ⅱgroups,mimic-NC group,and miR-101-3p mimic group.The relationship between miR-101-3p and the matrix metallopeptidase 15(MMP15)gene was verified by dual-luciferase reporter assay.The expression of miR-101-3p in cells was determined by quantitative real-time PCR.Cell proliferation,migration,invasion,and apoptosis were measured by MTT assay,scratch assay,Transwell chamber assay,and flow cytometry,respectively.Enzyme-linked immunosorbent assay kits were used to determine the levels of interleukin-6(IL-6),IL-1β,tumor necrosis factor-α(TNF-α),malondialdehyde(MDA),and glutathione peroxidase(GSH-Px)in cells and the release rate of lactate dehydrogenase(LDH).Western blot was used to measure the protein expression of proliferating cell nuclear antigen(PCNA),B-cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax),and MMP15.Results MMP15 was the target gene of miR-101-3p.Compared with the control group,the expression level of miR-101-3p,apoptosis rate,IL-6,IL-1β,TNF-α,the levels of LDH release,MDA levels,and Bax protein expression in the ox-LDL group were significantly increased(F=100.7-526.1,P<0.05),while the A value of cell proliferation,scratch healing rate,invasion number,GSH-Px activity,PCNA,Bcl-2,and MMP15 protein expression levels were significantly reduced(F=93.0-260.6,P<0.05).Compared with the ox-LDL group,the expression level of miR-101-3p,apoptosis rate,IL-6,IL-1β,TNF-α,the levels of LDH release rate,MDA levels,and Bax protein expression in the low,medium,and high dose groups of P-Ⅱwere significantly reduced(P<0.05),while the A value of cell proliferation,scratch healing rate,invasion number,GSH-Px activity,PCNA,Bcl-2,and MMP15 protein expression levels were significantly increased(P<0.05).Upregulation of miR-101-3p expression can weake

关 键 词：胡黄连苷Ⅱ 内皮 血管 动脉粥样硬化 氧化性应激 炎症 脂蛋白类 LDL 

分 类 号：R285.5[医药卫生—中药学] R543.5[医药卫生—中医学]