您的位置：山东大学 -> 科技期刊社 -> 《山东大学学报（医学版）》

山东大学学报 (医学版) ›› 2024, Vol. 62 ›› Issue (3): 11-19.doi: 10.6040/j.issn.1671-7554.0.2023.1124

• 基础医学 • 上一篇    下一篇

NR4A1通过IκBα/NF-κB通路调控过氧化氢诱导人脐静脉内皮细胞凋亡的机制

姜子晗1,芦兴晨1,孙露2,赵蕙琛3,左丹4,马小莉3,刘元涛5,张玉超4   

  1. 1.青岛大学附属青岛临床医学院 青岛市市立医院, 山东 青岛 266011;2.山东大学齐鲁医院(青岛)临床营养科, 山东 青岛 266011;3.青岛市市立医院内分泌科, 山东 青岛 266011;4.青岛市市立医院医学杂志编辑部, 山东 青岛 266011;5.山东大学齐鲁医院(青岛)内分泌科, 山东 青岛 266011
  • 发布日期:2024-05-06
  • 通讯作者: 张玉超. E-mail:yc.zhang@hotmail.com
  • 基金资助:
    山东省自然科学基金面上项目(ZR2022MH086);山东省医药卫生科技发展计划(202103060651);2021年度青岛市医药卫生科研计划(2021-WJZD005)

Mechanism of NR4A1 regulating hydrogen peroxide-induced apoptosis in human umbilical vein endothelial cells via the IκBα/NF-κB pathway

JIANG Zihan1, LU Xingchen1, SUN Lu2, ZHAO Huichen3, ZUO Dan4, MA Xiaoli3, LIU Yuantao5, ZHANG Yuchao4   

  1. 1. Qingdao Clinical Medical School of Qingdao University, Qingdao Municipal Hospital, Qingdao 266011, Shandong, China;
    2. Department of Clinical Nutrition, Qilu Hospital of Shandong University(Qingdao), Qingdao 266011, Shandong, China;
    3. Department of Endocrinology, Qingdao Municipal Hospital, Qingdao 266011, Shandong, China;
    4. Editorial Office of Medical Journal, Qingdao Municipal Hospital, Qingdao 266011, Shandong, China;
    5. Department of Endocrinology, Qilu Hospital of Shandong University(Qingdao), Qingdao 266011, Shandong, China
  • Published:2024-05-06

PDF (PC)

19

摘要： 目的 探讨过氧化氢(H2O2)诱导的人脐静脉内皮细胞(human umbilical vein endothelial cells, HUVECs)氧化应激中孤核受体NR4A1表达变化,以及其对细胞凋亡的影响及机制。 方法 使用不同浓度及时间梯度H2O2处理HUVECs,采用CCK-8法检测细胞活性,TUNEL法检测细胞凋亡,Western blotting法检测Bcl-2、Bax与NR4A1蛋白表达;采用siRNA转染建立敲低NR4A1与对照的HUVECs,分为si-NC组及si-NR4A1组,H2O2处理后检测各组细胞抑凋亡蛋白Bcl-2、凋亡蛋白Bax表达,TUNEL法检测细胞凋亡;采用慢病毒感染建立稳定过表达NR4A1与对照的HUVECs,并进行H2O2处理,分为空载对照组(NC组)、过表达组(OV组)、NC+H2O2组及OV+H2O2组。TUNEL法检测各组细胞凋亡、Western blotting法检测各组中NR4A1、Bcl-2、Bax、总IκBα的蛋白表达、P65蛋白在细胞核/质定位。 结果 200 μmol/L H2O2处理细胞6 h,HUVECs活力显著下降,凋亡水平显著上升,Bax/Bcl-2比值升高(P<0.001);NR4A1蛋白表达量增加;与si-NC组相比,si-NR4A1组在H2O2处理后细胞凋亡率、Bax/Bcl-2比值下降(P<0.001);OV+H2O2组细胞凋亡率、Bax/Bcl-2比值较NC+H2O2组显著降低(P<0.001);相较于NC+H2O2组,P65蛋白在OV+H2O2组中细胞核内表达显著降低(P<0.05),细胞质内表达显著上调(P<0.001);与NC+H2O2组相比,OV+H2O2组总IκBα表达上调(P<0.001)。 结论 H2O2诱导HUVECs发生细胞凋亡;HUVECs内NR4A1蛋白过表达可抑制细胞凋亡,其作用机制可能与调控IκBα的表达与抑制NF-κB的核转位有关。

关键词: 孤核受体NR4A1, 人脐静脉内皮细胞, IκBα蛋白, NF-κB核转位, 细胞凋亡

Abstract: Objective To investigate the expression of orphan nuclear receptor 4A1(NR4A1)in human umbilical vein endothelial cells(HUVECs)induced by hydrogen peroxide(H2O2)oxidative stress, and its effects and mechanism in cell apoptosis. Methods After HUVECs were treated with different concentrations and exposure times of H2O2, cell activity, apoptosis and protein expressions of Bcl-2, Bax and NR4A1 were detected with CCK-8, TUNEL and Western blotting, respectively. siRNA was transfected into HUVECs to obtain the knocked down NR4A1 cells(si-NR4A1)and control(si-NC). After H2O2 treatment, the levels of cell apoptosis, and the Bax/Bcl-2 ratio were detected. Lentivirus transfection was used to establish stably overexpressed NR4A1 HUVECs, which were treated with H2O2 and divided into empty vector control group(NC), NR4A1 overexpressed group(OV), NC+H2O2 group, and OV+H2O2 group. The cell apoptosis was determined with TUNEL, and the protein expressions of NR4A1, Bcl-2, Bax, total IκBα, and the nuclear/cytoplasmic localization of P65 protein in each group were determined with Western blotting. Results After the cells were treated with 200 μmol/L H2O2 for 6 hours, the vitality of HUVECs significantly decreased, the rate of apoptosis significantly increased, the Bax/Bcl-2 ratio increased(P<0.001), and the protein expression of NR4A1 increased. Compared with the si-NC group, the si-NR4A1 group had decreased Bax/Bcl-2 ratio and cell apoptosis after H2O2 treatment(P<0.001). Compared with the NC+H2O2 group, the OV+H2O2 group had significantly decreased cell apoptosis rate and Bax/Bcl-2 ratio(P<0.05). Compared with the NC+ H2O2 group, the OV+ H2O2 group had significantly decreased P65 protein expression in the nucleus(P<0.05), but significantly increased expression in the cytoplasm(P<0.001). Compared with the NC+ H2O2 group, the OV+ H2O2 group had significantly upregulated total IκBα expression(P<0.001). Conclusion H2O2 induces apoptosis in HUVECs; NR4A1 inhibits cell apoptosis by regulating the expression of IκBα and inhibiting the nuclear translocation of NF-κB in HUVECs.

Key words: Orphan nuclear receptor NR4A1, Human umbilical vein endothelial cells, IκBα protein, NF-κB nuclear translocation, Apoptosis

中图分类号: 

  • R587
[1] Sun H, Saeedi P, Karuranga S, et al. IDF diabetes atlas: global, regional and country-level diabetes prevalence estimates for 2021 and projections for 2045[J]. Diabetes Res Clin Pract, 2022, 183: 109119. doi: 10.1016/j.diabres.2021.109119.
[2] Viigimaa M, Sachinidis A, Toumpourleka M, et al. Macrovascular complications of type 2 diabetes mellitus[J]. Curr Vasc Pharmacol, 2020, 18(2): 110-116.
[3] Vorotnikov AV, Khapchaev AY, Nickashin AV, et al. In vitro modeling of diabetes impact on vascular endothelium: are essentials engaged to tune metabolism?[J]. Biomedicines, 2022, 10(12): 3181. doi: 10.3390/biomedicines10123181.
[4] Baaten CCFMJ, Vondenhoff S, Noels H. Endothelial cell dysfunction and increased cardiovascular risk in patients with chronic kidney disease[J]. Circ Res, 2023, 132(8): 970-992.
[5] Wang H, Zhang M, Fang F, et al. The nuclear receptor subfamily 4 group A1 in human disease[J]. Biochem Cell Biol, 2023, 101(2): 148-159.
[6] Hamers AA, Hanna RN, Nowyhed H, et al. NR4A nuclear receptors in immunity and atherosclerosis[J]. Curr Opin Lipidol, 2013, 24(5): 381-385.
[7] Cui M, Liu J, Geng L, et al. Let-7a Targeting TNFAPI3 promotes vascular endothelial cell apoptosis of pediatric patients with henoch-schönlein purpura via NF-κB signaling pathway[J]. J Healthc Eng, 2022, 2022: 3889318. doi: 10.1155/2022/3889318.
[8] Hu Y, Zhan Q, Liu HX, et al. Accelerated partial hepatectomy-induced liver cell proliferation is associated with liver injury in Nur77 knockout mice[J]. Am J Pathol, 2014, 184(12): 3272-3283.
[9] Tian H, Chen F, Wang Y, et al. Nur77 prevents osteoporosis by inhibiting the NF-κB signalling pathway and osteoclast differentiation[J]. J Cell Mol Med, 2022, 26(8): 2163-2176.
[10] Li L, Liu Y, Chen HZ, et al. Impeding the interaction between nur77 and p38 reduces LPS-induced inflammation[J]. Nat Chem Biol, 2015, 11(5): 339-346.
[11] Jing Cao, Zhang G, Liu Z, et al. Peroxidasin promotes diabetic vascular endothelial dysfunction induced by advanced glycation end products via NOX2/HOCl/Akt/eNOS pathway[J]. Redox Biol, 2021, 45: 102031. doi: 10.1016/j.redox.2021.102031.
[12] 扈艳雯, 赵蕙琛, 马小莉, 等. GLP-1通过细胞色素P450表氧化酶途径抑制氧化应激[J]. 山东大学学报(医学版), 2023, 61(8): 10-16. HU Yanwen, ZHAO Huichen, MA Xiaoli, et al. GLP-1 inhibits oxidative stress damage through cytochrome P450 surface oxidase pathway[J]. Journal of Shandong University(Health Science), 2023, 61(8): 10-16.
[13] Herring JA, Elison WS, Tessem JS. Function of Nr4a orphan nuclear receptors in proliferation, apoptosis and fuel utilization across tissues[J]. Cells, 2019, 8(11): 1373.
[14] Chen L, Fan F, Wu L, et al. The nuclear receptor 4A family members: mediators in human disease and autophagy[J]. Cell Mol Biol Lett, 2020, 25(1): 48.
[15] Crean D, Murphy EP. Targeting NR4A nuclear receptors to control stromal cell inflammation, metabolism, angiogenesis, and tumorigenesis[J]. Front Cell Dev Biol, 2021, 9: 589770. doi: 10.3389/fcell.2021.589770.
[16] Deng S, Chen B, Huo J, et al. Therapeutic potential of NR4A1 in cancer: focus on metabolism[J]. Front Oncol, 2022, 12: 972984. doi: 10.3389/fonc.2022.972984.
[17] Bending D, Zikherman J. Nr4a nuclear receptors: markers and modulators of antigen receptor signaling[J]. Curr Opin Immunol, 2023, 81: 102285. doi: 10.1016/j.coi.2023.102285.
[18] Zheng Y, Tao Y, Zhan X, et al. Nuclear receptor 4A1(NR4A1)silencing protects hepatocyte against hypoxia-reperfusion injury in vitro by activating liver kinase B1(LKB1)/AMP-activated protein kinase(AMPK)signaling[J]. Bioengineered, 2022, 13(4): 8349-8359.
[19] Yu C, Cui S, Zong C, et al. The orphan nuclear receptor NR4A1 protects pancreatic β-Cells from endoplasmic reticulum(ER)stress-mediated apoptosis[J]. J Biol Chem, 2015, 290(34): 20687-20699.
[20] Ding R, Sun X, Yi B, et al. Nur77 Attenuates inflammasome activation by inhibiting caspase-1 expression in pulmonary vascular endothelial cells[J]. Am J Respir Cell Mol Biol, 2021, 65(3): 288-299.
[21] Popichak KA, Hammond SL, Moreno JA, et al. Compensatory expression of nur77 and nurr1 regulates nf-κb-dependent inflammatory signaling in astrocytes[J]. Mol Pharmacol, 2018, 94(4): 1174-1186.
[22] Freire PR, Conneely OM. NR4A1 and NR4A3 restrict HSC proliferation via reciprocal regulation of C/EBPα and inflammatory signaling[J]. Blood, 2018, 131(10): 1081-1093.
[23] Barnabei L, Laplantine E, Mbongo W, et al. NF-κB: at the borders of autoimmunity and inflammation[J]. Front Immunol, 2021, 12: 716469. doi: 10.3389/fimmu.2021.716469.
[24] Bian W, Jing X, Yang Z, et al. Downregulation of LncRNA NORAD promotes Ox-LDL-induced vascular endothelial cell injury and atherosclerosis[J]. Aging, 2020, 12(7): 6385-6400.
[25] Liang WJ, Yang HW, Liu HN, et al. HMGB1 upregulates NF-kB by inhibiting IKB-α and associates with diabetic retinopathy[J]. Life Sci, 2020, 241: 117146. doi: 10.1016/j.lfs.2019.117146.
[1] 王晓磊 张海涛 张辉 郭成浩. 舒血宁注射液对高碘致培养血管内皮细胞损伤的保护作用[J]. 山东大学学报(医学版), 2209, 47(6): 38-.
[2] 鹿向东 杨伟 徐广明 曲元明. 脑膜瘤中PPAR-γ的表达及曲格列酮对脑膜瘤培养细胞生长的影响[J]. 山东大学学报(医学版), 2209, 47(6): 65-.
[3] 刘金波,刘凯文,向崇鑫,程雷. 西红花苷对椎间盘退变的保护作用[J]. 山东大学学报 (医学版), 2023, 61(9): 84-93.
[4] 扈艳雯,赵蕙琛,马小莉,刘元涛,张玉超. GLP-1通过细胞色素P450表氧化酶途径抑制氧化应激[J]. 山东大学学报 (医学版), 2023, 61(8): 10-16.
[5] 杨元凤,熊高才,黎豫川,罗玉玲,张敬杰. 鹿苓安肾颗粒对慢性肾功能衰竭大鼠炎症反应及细胞凋亡的影响[J]. 山东大学学报 (医学版), 2023, 61(10): 9-16.
[6] 赵舸,邹存华,宋冬冬,赵淑萍. 丹参酮IIA对子宫内膜癌细胞增殖与凋亡的影响[J]. 山东大学学报 (医学版), 2022, 60(9): 53-58.
[7] 刘敏,张玉超,马小莉,刘昕宇,孙露,左丹,刘元涛. 孤核受体NR4A1在H2O2诱导小鼠肾脏足细胞损伤中的作用[J]. 山东大学学报 (医学版), 2022, 60(5): 16-21.
[8] 封海岗,刘国文,曹洪. 干扰MAD2L1基因表达对乳腺癌细胞凋亡的影响及机制[J]. 山东大学学报 (医学版), 2022, 60(10): 9-16.
[9] 卢游,且迪,伍晋辉,杨凡. 干预Sonic Hedgehog信号通路对宫内发育迟缓新生大鼠学习记忆能力的影响[J]. 山东大学学报 (医学版), 2021, 59(5): 82-89.
[10] 薛源,林雪艳,徐歌,田永杰. 低氧诱导因子-1α在子宫内膜异位症患者血清中的表达和对在位子宫内膜间质细胞上皮-间质转化的影响[J]. 山东大学学报 (医学版), 2021, 59(2): 41-47.
[11] 张晓璐,王丽莉,陈凯明,娄宪芝,张曼. 组蛋白去乙酰化酶SIRT1经Toll样受体4途径对巨噬细胞凋亡的调控[J]. 山东大学学报 (医学版), 2020, 58(12): 8-14.
[12] 史丽,马静,赵喜娃,关英霞,赵连梅,单保恩. miR-25-3p在40例子宫内膜腺癌组织中的表达及对KLE细胞生物学功能的影响[J]. 山东大学学报 (医学版), 2020, 58(12): 86-91.
[13] 毕涛,卢倩倩,孙传东. 腺病毒介导HBx对人淋巴细胞影响的体外实验[J]. 山东大学学报 (医学版), 2018, 56(7): 15-20.
[14] 李孝峰,杜晓益,刘海南,刘承,范医东. 小檗碱对人肾细胞癌细胞增殖、凋亡、DNA断裂及损伤修复的影响[J]. 山东大学学报 (医学版), 2018, 56(3): 54-59.
[15] 陈希彦,王琪,顾伟亭,文勇. 干扰TAZ基因对舌鳞癌细胞CAL27增殖凋亡的影响及其机制[J]. 山东大学学报 (医学版), 2018, 56(10): 79-85.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
[1] 罗昕,何兵,聂清生,侯震波,董军,李玉花,曾祥芹,刘伟,孔德民,曹金凤. 磁共振扩散加权成像单指数模型与扩散峰度成像模型在61例肾透明细胞癌分级中的对比[J]. 山东大学学报 (医学版), 2020, 1(7): 89 -95 .
[2] 张迪,于猛,刘霞. 神经调控技术简述[J]. 山东大学学报 (医学版), 2020, 1(8): 50 -60 .
[3] 白洁,刘玥,张宁宁,温洋,彭芸,程华. 中枢神经肿瘤样脱髓鞘病2例[J]. 山东大学学报 (医学版), 2020, 1(9): 103 -105 .
[4] 王稳,董绍华,田翔宇,崔秀娟. 盆腹腔多发脾种植误诊为妇科肿瘤1例[J]. 山东大学学报 (医学版), 2020, 1(9): 116 -118 .
[5] 主余华,张春清,任万华,马艳丽,赵幼安. 发卡样CTGF特异性RNA干扰重组体的构建及筛选[J]. 山东大学学报(医学版), 2006, 44(4): 351 -356 .
[6] 孙维彤,邹伟伟,李爱国,席延伟,张娜. 脂质体粒径对促进托氟啶口服吸收的影响[J]. 山东大学学报(医学版), 2007, 45(6): 639 -642 .
[7] 汤树海,潘晓军. 阻塞性睡眠呼吸暂停综合征患者体内氧自由基代谢的作用[J]. 山东大学学报(医学版), 2006, 44(5): 475 -477 .
[8] 敏,李延青,刘传勇,谢冬萍,郭玉婷,左秀丽,曾娟. 肠易激综合征大鼠模型外周血免疫调节性T细胞的研究[J]. 山东大学学报(医学版), 2006, 44(5): 514 -518 .
[9] 慈倩倩，李梅，吴克良，马水英，王苗苗，刘辉，高芹，石玉华，陈子江. 玻璃化冷冻对人体外成熟卵母细胞微丝分布及发育潜能的影响[J]. 山东大学学报（医学版）, 2014, 52(4): 26 -29 .
[10] 杨洪超, 王健, 张文同, 李爱武. 先天性巨结肠症中Neuroligin2基因的表达及意义[J]. 山东大学学报（医学版）, 2014, 52(9): 15 -19 .
版权所有 © 2018 《山东大学学报(医学版)》编辑部　
地址:山东大学科技期刊社（济南市历城区山大南路27号山东大学中心校区明德楼B座721室） 电话: 0531-88366918 E-mail: xbyxb@sdu.edu.cn
本系统由北京玛格泰克科技发展有限公司设计开发