主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

北京中医药大学学报 ›› 2021, Vol. 44 ›› Issue (5): 413-420.doi: 10.3969/j.issn.1006-2157.2021.05.005

• 中药药理 • 上一篇    下一篇

白藜芦醇抑制Gly-LDL诱导人脐静脉内皮细胞损伤的作用机制研究*

刘美志1, 高洁2, 沙雯君2, 孙杜桑1, 邱军辉3, 雷涛1,2#   

  1. 1 上海中医药大学附属普陀医院 上海 200062;
    2 上海市普陀区中心医院内分泌科;
    3 安徽医科大学
  • 收稿日期:2020-10-27 出版日期:2021-05-30 发布日期:2021-06-01
  • 通讯作者: #雷涛,男,博士,教授,主任医师,博士生导师,主要研究方向:中西医结合防治糖尿病血管病变,E-mail:leitao5899@126.com
  • 作者简介:刘美志,女,在读硕士生
  • 基金资助:
    *国家自然科学基金项目(No.81774083),上海市医学重点专科建设项目(No.ZK2019B16),上海市普陀区卫生健康系统临床特色专科建设项目(No.2020tszk01)

Study on the inhibitory effect of resveratrol on injury of human umbilical vein endothelial cells*

Liu Meizhi1, Gao Jie2, Sha Wenjun2, Sun Dusang1, Qiu Junhui3, Lei Tao1,2#   

  1. 1 Putuo Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200062,China;
    2 Department of Endocrinology,Shanghai Putuo District Central Hospital,Shanghai 200062, China;
    3 Anhui Medical University, Anhui 230032,China
  • Received:2020-10-27 Online:2021-05-30 Published:2021-06-01
  • Contact: Prof. Lei Tao,Ph.D.,Chief Physician,Doctoral Supervisor. Putuo Hospital, Shanghai University of Traditional Chinese Medicine,164 Lanxi Road,Shanghai 200062. E-mail:leitao5899@126.com
  • Supported by:
    National Natural Science Foundation of China(No.81774083), Shanghai Key Medical Specialities (No.ZK2019B16),Clinical Characteristics of Health System in Putuo District, Shanghai(No.2020tszk01)

摘要: 目的 探究白藜芦醇(RSV)对糖化低密度脂蛋白(Gly-LDL)致人脐静脉内皮细胞(HUVECs)损伤的影响及作用机制。方法 体外培养人脐静脉内皮细胞,Gly-LDL诱导其损伤,设置为模型组;用不同浓度(5、10、20 μmol/L)白藜芦醇干预内皮细胞24 h,分别设置为白藜芦醇低浓度组、白藜芦醇中浓度组和白藜芦醇高浓度组;另设置空白组。通过CCK-8法检测细胞增殖能力,划痕实验观察细胞迁移能力,流式细胞术测定细胞凋亡水平,ELISA法检测内皮细胞中白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)的含量,Western blot法检测Toll样受体4(TLR4)、缺氧诱导因子-1α(HIF-1α)、IL-6和TNF-α的蛋白表达,实时荧光定量PCR法检测TLR4、HIF-1α、IL-6和TNF-α的mRNA表达。结果 与空白组比较,模型组细胞增殖和迁移能力下降(P<0.05, P<0.01),细胞凋亡率增加(P<0.01),血清IL-6和TNF-α含量增加(P<0.01),内皮细胞中TLR4、HIF-1α、IL-6和TNF-α的蛋白和mRNA表达均增加(P<0.05, P<0.01)。与模型组相比,白藜芦醇干预后,Gly-LDL损伤人脐静脉内皮细胞情况减轻,白藜芦醇中浓度组和白藜芦醇高浓度组的细胞增殖、迁移能力增强(P<0.05),细胞凋亡率下降(P<0.05),血清IL-6和TNF-α含量下降(P<0.05),内皮细胞中TLR4、HIF-1α、IL-6和TNF-α的蛋白表达和mRNA表达均减少(P<0.05)。结论 白藜芦醇能有效减轻Gly-LDL对人脐静脉内皮细胞的炎症损伤,增强细胞增殖、迁移能力,抑制细胞凋亡,其机制可能与白藜芦醇下调TLR4/HIF-1α信号通路有关。

关键词: 白藜芦醇, 糖化低密度脂蛋白, 人脐静脉内皮细胞, 信号通路

Abstract: Objective To explore the effect and mechanism of resveratrol on human umbilical vein endothelial cells injury induced by glycosylated low density lipoprotein (Gly-LDL).Methods Human umbilical vein endothelial cells were cultured in vitro.The model group was induced by Gly-LDL.The low-concentration, medium-concentration and high-concentration RSV groups were established by treating with 5 μmol/L, 10 μmol/L and 20 μmol/L resveratrol respectively for 24 hours.The cell proliferative capacity was detected by Cell Counting Kit-8 (CCK-8), the ability of cell migration was observed by scratch essay, the level of apoptosis was measured by flow cytometry, the contents of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in endothelial cells were detected by ELISA, and the protein expressions of Toll-like receptor 4 (TLR4), hypoxia-inducible factor-1 (HIF-1α), IL-6 and TNF-α were detected by Western-blot.The mRNA expressions of TLR4, HIF-1 α, IL-6 and TNF-α were detected by real-time fluorescence quantitative PCR.Results Compared with the blank group, the cell proliferative capacity and migration ability decreased (P<0.05, P<0.01),the rate of apoptosis increased (P<0.01),the contents of IL-6 and TNF-α in serum increased (P<0.01),and the protein expression and mRNA expression of TLR4, HIF-1α, IL-6 and TNF-α in endothelial cells increased in the model group (P<0.05, P<0.01). Compared with the model group, the injury of human umbilical vein endothelial cells induced by Gly-LDL was alleviated after the intervention of resveratrol. The ability of cell proliferation and migration was enhanced (P<0.05), the apoptosis rate was decreased (P<0.05), the contents of serum IL-6 and TNF-α were decreased (P<0.05), and the protein expression and mRNA expression of TLR4, HIF-1α, IL-6 and TNF-α in endothelial cells were decreased in the medium-concentration and high-concentration RSV groups(P<0.05).Conclusion Resveratrol seems to reduce the inflammatory injury of human umbilical vein endothelial cells induced by Gly-LDL, enhance the ability of cell proliferation and migration, and inhibit cellapoptosis, which may be related to the down-regulation of TLR4/HIF-1α signal pathway by resveratrol.

Key words: resveratrol, Gly-LDL, human umbilical vein endothelial cells, signal pathway

中图分类号: 

  • R285.5