GLP-1通过miR-22/NLRP3途径减轻ox-LDL诱导的内皮细胞焦亡
作者:
作者单位:

(台州市中西医结合医院, 浙江省台州市317523)

作者简介:

陈小兰,主治医师,研究方向为心内科相关疾病,E-mail为15305882668@189.cn。


GLP-1 alleviates ox-LDL-induced endothelial cell pyroptosis via miR-22/NLRP3 pathway
Author:
Affiliation:

Taizhou Integrated Chinese and Western Medicine Hospital, Taizhou, Zhejiang 317523, China)

  • 摘要
  • | |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • | |
    摘要:

    目的 探讨胰高血糖素样肽1(GLP-1)对氧化型低密度脂蛋白(ox-LDL)诱导人脐静脉内皮细胞(HUVEC)焦亡的调节作用及分子机制。方法 培养HUVEC并分为对照组、ox-LDL组、不同剂量GLP-1组(10 nmol/L、100 nmol/L、1 000 nmol/L)、GLP-1+miR-22抑制物组、miR-22抑制物组、miR-22模拟物组、阴性对照(NC)抑制物组、阴性对照模拟物组。流式细胞术检测细胞凋亡率,荧光定量PCR检测miR-22表达量,Western blot检测NOD样受体蛋白3(NLRP3)、凋亡相关点样蛋白(ASC)、含半胱氨酸的天冬氨酸蛋白水解酶1(Caspase-1)的表达量,酶联免疫吸附法检测白细胞介素1β(IL-1β)、IL-18的含量。结果 与对照组比较,ox-LDL组细胞凋亡率和细胞中NLRP3、ASC、Caspase-1的表达以及培养基中IL-1β、IL-18含量均明显增加,细胞中miR-22的表达明显减少(P<0.05);与ox-LDL组比较,GLP-1组细胞凋亡率和细胞中NLRP3、ASC、Caspase-1的表达以及培养基中IL-1β、IL-18含量均明显减少,细胞中miR-22的表达明显增加(P<0.05);与GLP-1组比较,GLP-1+miR-22抑制物组细胞凋亡率和细胞中NLRP3、ASC、Caspase-1的表达以及培养基中IL-1β、IL-18的含量均明显增加,细胞中miR-22的表达明显减少(P<0.05);miR-22抑制物组细胞中NLRP3、ASC、Caspase-1的表达及培养基中IL-1β、IL-18的含量明显高于阴性对照抑制物组,miR-22模拟物组细胞中NLRP3、ASC、Caspase-1的表达及培养基中IL-1β、IL-18的含量明显低于阴性对照模拟物组(P<0.05)。结论 GLP-1通过miR-22/NLRP3途径能够减轻ox-LDL诱导的内皮细胞焦亡。

    Abstract:

    Aim To study the regulatory effect and molecular mechanism of glucagon-like peptide-1 (GLP-1) on pyroptosis of umbilical vein endothelial cells (HUVEC) induced by oxidized low density lipoprotein (ox-LDL). Methods HUVEC was cultured and divided into control group, ox-LDL group, GLP-1 group (10 nmol/L, 100 nmol/L, 1 000 nmol/L), GLP-1+miR-22 inhibitor group, miR-22 inhibitor group, miR-22 mimic group, negative control (NC) inhibitor group and NC mimic group. Flow cytometry was used to detect apoptotic rate, fluorescence quantitative PCR was used to detect the expression of microRNA-22(miR-22), Western blot was used to detect the expression of NOD-like receptor protein 3 (NLRP3), apoptosis-associated speck-like protein containing CARD (ASC) and caspase-1, enzyme-linked immunosorbent assay was used to detect the content of interleukin 1beta (IL-1β) and interleukin-18(IL-18). Results Compared with the control group, the apoptotic rate, the expression of NLRP3, ASC, caspase-1 in cells and the levels of IL-1β and IL-18 in the culture medium of ox-LDL group significantly increased, while the expression of miR-22 significantly decreased (P<0.05). Compared with ox-LDL group, the apoptotic rate, the expression of NLRP3, ASC, caspase-1 in cells and the levels of IL-1β and IL-18 in the culture medium of GLP-1 group significantly decreased, while the expression of miR-22 significantly increased (P<0.05). Compared with GLP-1 group, the apoptotic rate, the expression of NLRP3, ASC, caspase-1 in cells and the levels of IL-1β and IL-18 in the culture medium of GLP-1+miR-22 inhibitor group significantly increased, while the expression of miR-22 significantly decreased (P<0.05). The expression of NLRP3, ASC, caspase-1 in cells and the levels of IL-1β and IL-18 in the culture medium of miR-22 inhibitor group were significantly higher than those of NC inhibitor group, while the expression of NLRP3, ASC, caspase-1 in cells and the levels of IL-1β and IL-18 in the culture medium of miR-22 mimic group were significantly higher than those of NC mimic group. Conclusion GLP-1 can alleviate ox-LDL-induced endothelial cell pyroptosis through the miR-22/NLRP3 pathway.

    参考文献
    [1] Gallo G, Pierelli G, Forte M, et al.Role of oxidative stress in the process of vascular remodeling following coronary revascularization.Int J Cardiol, 8,1(268):27-33.
    [2] Sun MY, Zhang M, Chen SL, et al.The influence of hyperlipidemia on endothelial function of FPN1 Tek-Cre mice and the intervention effect of tetramethylpyrazine.Cell Physiol Biochem, 8,7(1):119-128.
    [3] Karki P, Birukov KG.Lipid mediators in the regulation of endothelial barriers.Tissue Barriers, 8,6(1):e1385573.
    [4] Lin F, Pei L, Zhang Q, et al.Ox-LDL induces endothelial cell apoptosis and macrophage migration by regulating caveolin-1 phosphorylation.J Cell Physiol, 8,3(10):6683-6692.
    [5] Zhang M, Jiang L.Oxidized low-density lipoprotein decreases VEGFR2 expression in HUVECs and impairs angiogenesis.Exp Ther Med, 6,2(6):3742-3748.
    [6] Cai X, She M, Xu M, et al.GLP-1 treatment protects endothelial cells from oxidative stress-induced autophagy and endothelialdysfunction.Int J Biol Sci, 8,4(12):1696-1708.
    [7] 高维鸿, 蔡祥胜, 谭照光, 等.胰高血糖素样肽-1改善过氧化氢诱导的血管内皮氧化损伤.暨南大学学报(自然科学与医学版), 7,8(1):56-62.
    [8] 孙慧琳, 黄志秋, 曾海龙, 等.GLP-1通过NOX4信号通路拮抗AGE诱导的人脐静脉内皮细胞氧化损伤.中山大学学报(医学科学版), 6,7(2):197-1,9.
    [9] Lara-Guzmán OJ, Gil-Izquierdo , Medina S, et al.Oxidized LDL triggers changes in oxidative stress and inflammatory biomarkers in human macrophages.Redox Biol, 8,5:1-11.
    [10] Fu Z, Zhou E, Wang X, et al.Oxidized low-density lipoprotein-induced microparticles promote endothelial monocyte adhesion via intercellular adhesion molecule 1.Am J Physiol Cell Physiol, 7,3(5):567-574.
    [11] Cheang JY, Moyle PM.Glucagon-like peptide-1 (GLP-1)-based therapeutics:current status and future opportunities beyond type 2 diabetes.Chem Med Chem, 8,3(7):662-671.
    [12] Lim DM, Park KY, Hwang WM, et al.Difference in protective effects of GIP and GLP-1 on endothelial cells according to cyclic adenosine monophosphate response.Exp Ther Med, 7,3(5):2558-2564.
    [13] Wang Y, Liu J, Chen X, et al.Dysfunctional endothelial-derived microparticles promote inflammatory macrophage formation via NF-кB and IL-1β signal pathways.J Cell Mol Med, 9,3(1):476-486.
    [14] Kaniewska-Bednarczuk E, Mielcarek M, Chester AH, et al.Oxidized low-density lipoproteins enhance expression and activity of CD39 and CD73 in the human aortic valve endothelium.Nucleosides Nucleotides Nucleic Acids, 6,5(10-12):713-719.
    [15] Huang WQ, Wei P, Lin RQ, et al.Protective effects of microRNA-22 against endothelial cell injury by targeting NLRP3 through suppression of the inflammasome signaling pathway in a rat model of coronary heart disease.Cell Physiol Biochem, 7,3(4):1346-1358.
    [16] Grebe A, Hoss F, Latz E.NLRP3 inflammasome and the IL-1 pathway in atherosclerosis.Circ Res, 8,2(12):1722-1740.
    [17] Koka S, Xia M, Chen Y, et al.Endothelial NLRP3 inflammasome activation and arterial neointima formation associated with acid sphingomyelinase during hypercholesterolemia.Redox Biol, 7,3:336-344.
    [18] 胡波, 张晓刚, 李德才.miR-22靶向抑制NLRP3基因对冠心病内皮细胞炎症损伤的保护作用.安徽医科大学学报, 8,3(5):668-675.
    引证文献
引用本文

陈小兰,陶福正,郑道国,吴利云. GLP-1通过miR-22/NLRP3途径减轻ox-LDL诱导的内皮细胞焦亡[J].中国动脉硬化杂志,2019,27(5):410~416.

复制
分享
文章指标
  • 点击次数:1578
  • 下载次数: 874
历史
  • 收稿日期:2019-01-16
  • 最后修改日期:2019-03-05
  • 在线发布日期: 2019-04-08