摘要
目的探究芹菜素(AP)对高糖高脂(HG/HF)应激造成的血管内皮细胞功能失调的修复作用及机制。方法将人体脐静脉内皮细胞(HUVECs)分为6组:对照组不经任何处理;A组经HG/HF处理;B组经HG/HF和5μM AP处理;C组经HG/HF和25μM AP处理;D组经HG/HF和50μM AP处理;E组经HG/HF、25μM AP以及PI3K抑制剂LY294002(10μM)处理。用细胞划痕实验评估各组愈合修复能力;用CCK8法测定各组增殖能力;用蛋白免疫印迹检测各组PI3K-Akt通路中蛋白表达情况;用酶联免疫吸附测定法(ELISA)和生化试剂盒测定各组培养基内皮素-1(ET-1)和NO水平;用流式细胞术测定各组细胞内活性氧(ROS)水平;用qRT-PCR检测各组肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)mRNA表达水平。结果C组和D组细胞迁移率显著明显高于A组(P<0.05);D组细胞增殖率显著明显高于A组(P<0.05);B组、C组和D组细胞内ROS水平均显著低于A组(P<0.05)。与A组相比,C组和D组细胞ET-1水平明显降低,NO水平明显升高(均P<0.05);B组、C组和D组IL-6 mRNA表达水平明显下调(均P<0.05),C组和D组TNF-αmRNA表达水平明显下调(P<0.05);C组和D组p-PI3K和p-AKT的水平明显升高(均P<0.05)。E组细胞培养液中ET-1水平明显高于D组,而NO水平明显低于D组(均P<0.05),且E组IL-6和TNF-αmRNA表达水平较D组明显上调(均P<0.05)。结论AP具有改善HG/HF应激造成的血管内皮细胞功能失调的作用,其作用依赖于PI3K-Akt通路的激活。
Objective To investigate the restorative effect of apigenin(AP)on vascular endothelial cell dysfunction induced by high glucose/high fat(HG/HF)stress and its possible mechanism.Methods Human umbilical vein endothelial cells(HUVECs)were divided into 6 groups:the control group received no treatment;group A was treated with HG/HF;group B was treated with HG/HF and 5μM AP;group C was treated with HG/HF and 25μM AP;group D was treated with HG/HF and 50μM AP;group E was treated with HG/HF,25μM AP,and the PI3K inhibitor LY294002(10μM).In each group,the effect of wound healing and repair were evaluated using the cell scratch assay;cell proliferation was determined using CCK8;the expression of PI3K-Akt pathway protein was detected by Western blotting;the levels of endothelin-1(ET-1)and NO in cell culture medium were measured using enzyme-linked immunosorbent assay(ELISA)and biochemical assay kits;the reactive oxygen species(ROS)levels were determined by flow cytometry;interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)mRNA levels were detected using qRT-PCR.Results The cell migration rates of groups C and D were significantly higher than that of group A(P<0.05);the cell proliferation rate of group D was significantly higher than that of group A(P<0.05);the intracellular ROS levels in groups B,C,and D were all significantly lower than that of group A(P<0.05).Compared with group A,groups C and D showed significantly reduced ET-1 levels and significantly increased NO levels(both P<0.05);groups B,C,and D demonstrated significant downregulation of IL-6 mRNA expression levels(all P<0.05),while groups C and D exhibited significant downregulation of TNF-αmRNA expression levels(P<0.05);the levels of p-PI3K and p-AKT were significantly elevated in groups C and D(both P<0.05).The ET-1 level in the cell culture medium of group E was significantly higher than that of group D,while the NO level was significantly lower(both P<0.05),and the IL-6 and TNF-αmRNA expression levels in group E were significantly upregulated compared
作者
熊绍风
李炜
邱雪峰
王昆
严国强
赵乐
方洋
XIONG Shao-feng;LI Wei;QIU Xue-feng;WANG Kun;YAN Guo-qiang;ZHAO Le;FANG Yang(Department of Pharmacy,the First Affiliated Hospital of Nanchang University,Nanchang 330006,China;Department of Pharmacy,Jiangxi Cancer Hospital,Nanchang 330029,China)
出处
《南昌大学学报（医学版）》
2024年第6期27-32,72,共7页
Journal of Nanchang University：Medical Sciences
基金
江西省医院药学专项研究基金资助项目(2024-ZXYJ01)
江西省中医药管理局科技计划重点项目(2020Z016)。
