黄嘌呤氧化还原酶在血管内皮细胞缺氧损伤硝酸盐一氧化氮转化中的作用探讨

doi:10.19438/j.cjoms.2021.04.001

中国口腔颌面外科杂志 ›› 2021, Vol. 19 ›› Issue (4): 289-294.doi: 10.19438/j.cjoms.2021.04.001

黄嘌呤氧化还原酶在血管内皮细胞缺氧损伤硝酸盐一氧化氮转化中的作用探讨

杨扬, 牛其芳, 李德龙, 张姝, 秦力铮, 韩正学

首都医科大学附属北京口腔医院口腔颌面-头颈肿瘤科,北京 100050

收稿日期:2020-12-28 修回日期:2021-03-11 出版日期:2021-07-20 发布日期:2021-08-05
通讯作者: 韩正学,E-mail：hanf1989@hotmail.com
作者简介:杨扬（1991-）,男,博士,医师,E-mail：yangsun910@ccmu.edu.cn
基金资助:
国家自然科学基金（81974144、81570957）; 首都医科大学附属北京口腔医院青年科技创新人才培育计划（YSP202008）; 北京口腔医院学科建设基金（18-09-21）

The role of xanthine oxidoreductase in nitrate-nitric oxide transformation in vascular endothelial cells of hypoxia injury

YANG Yang, NIU Qi-fang, LI Delong, ZHANG Shu, QIN Li-zheng, HAN Zheng-xue

Department of Oral and Maxillofacial Head and Neck Oncology, Beijing Stomatological Hospital, Capital Medical University. Beijing 100050, China

Received:2020-12-28 Revised:2021-03-11 Online:2021-07-20 Published:2021-08-05

摘要/Abstract

摘要： 目的： 探讨缺氧状态下的血管内皮细胞硝酸盐转化生成一氧化氮（NO）的机制及黄嘌呤氧化还原酶(XOR)在其中的作用。方法： 通过人血管内皮细胞缺氧复氧损伤模型,细胞使用含不同浓度的硝酸钠和亚硝酸钠培养基培养,检测血管内皮细胞内硝酸钠含量,使用NO探针检测NO水平,实时定量PCR及免疫印迹实验检测内皮源性一氧化氮合成酶(eNOS)和XOR的表达。使用相关抑制剂L-NMMA和别嘌呤醇（allopurinol）后,再次检测NO生成水平。采用GraphPad Prism 8.0软件包进行统计学分析。结果： 较高浓度的细胞外硝酸盐在常氧和缺氧条件下都会增加细胞内硝酸盐含量,细胞外一定浓度的亚硝酸盐和硝酸盐仅在缺氧条件下会增加细胞内NO水平。使用XOR阻断剂别嘌呤醇后,细胞外硝酸盐不再使细胞内NO水平升高。结论： 缺氧状态下,血管内皮细胞启动硝酸盐NO转化,XOR可能是这一过程中的关键因素。

关键词: 血管内皮细胞, 缺氧损伤, 硝酸盐-亚硝酸盐-一氧化氮途径, 黄嘌呤氧化还原酶

Abstract: PURPOSE: To investigate the mechanism of nitrate-nitric oxide (NO) transformation in vascular endothelial cells under hypoxia injury and the role of xanthine oxidoreductase(XOR). METHODS: Model of hypoxia reoxygenation injury of human vascular endothelial cells was established. The level of nitric oxide and the content of sodium nitrate were detected in cells cultured in medium with different concentrations of sodium nitrate and sodium nitrite. The expression of endothelial nitric oxide synthase(eNOS) and XOR were detected by real-time quantitative PCR and Western blot. After eNOS inhibitor L-NMMA and XOR inhibitor allopurinol were used respectively, the levels of NO in cells cultured in medium with sodium nitrate were detected again. The data were analyzed with GraphPad Prism 8.0 software package. RESULTS: Higher concentration of extracellular nitrate increased intracellular nitrate content in both normoxia and hypoxia conditions. Nitrite and nitrate extracellular only increased levels of intracellular NO in hypoxia condition, and with allopurinol, XOR inhibitor, extracellular nitrate did not increase intracellular NO level. CONCLUSIONS: Nitrate-NO transformation in human vascular endothelial cells occurs in condition of hypoxia, which is mainly dependent on XOR.

Key words: Vascular endothelial cells, Hypoxia injury, Nitrate-nitrite-nitric oxide pathway, Xanthine oxidoreductase

中图分类号:

杨扬, 牛其芳, 李德龙, 张姝, 秦力铮, 韩正学. 黄嘌呤氧化还原酶在血管内皮细胞缺氧损伤硝酸盐一氧化氮转化中的作用探讨[J]. 中国口腔颌面外科杂志, 2021, 19(4): 289-294.

YANG Yang, NIU Qi-fang, LI Delong, ZHANG Shu, QIN Li-zheng, HAN Zheng-xue. The role of xanthine oxidoreductase in nitrate-nitric oxide transformation in vascular endothelial cells of hypoxia injury[J]. China J Oral Maxillofac Surg, 2021, 19(4): 289-294.

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黄嘌呤氧化还原酶在血管内皮细胞缺氧损伤硝酸盐一氧化氮转化中的作用探讨

The role of xanthine oxidoreductase in nitrate-nitric oxide transformation in vascular endothelial cells of hypoxia injury

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