褪黑素抑制H2O2诱导的人脐静脉内皮细胞氧化应激及凋亡标志蛋白质的表达

doi:10.16352/j.issn.1001-6325.2024.05.0645

基础医学与临床 ›› 2024, Vol. 44 ›› Issue (5): 645-650.doi: 10.16352/j.issn.1001-6325.2024.05.0645

褪黑素抑制H₂O₂诱导的人脐静脉内皮细胞氧化应激及凋亡标志蛋白质的表达

杨瀚毅^1,2#, 宁佳怡^1,2#, 汪小兰^2#, 张益萌², 谢婷珂^1,2, 陈奕玄^1,2, 韩静^2*

1.西安医学院眼科学教研室,陕西西安 710068;
2.空军军医大学第二附属医院眼科, 陕西西安 710038

收稿日期:2023-09-26 修回日期:2024-01-24 出版日期:2024-05-05 发布日期:2024-04-23
通讯作者: ^*hanjing.cn@163.com
^#对本文有相同贡献
基金资助:
国家自然科学基金 (8217111370);陕西省科技攻关重点研发计划 (2021SF-158);唐都医院临床重点研究 (2021LCYJ019,2018LCYJ008)

Melatonin inhibits H₂O₂-induced oxidative stress and apoptosis marker protein expression in human umbilical vein endothelial cells

YANG Hanyi^1,2#, NING Jiayi^1,2#, WANG Xiaolan^2#, ZHANG Yimeng², XIE Tingke^1,2, CHEN Yixuan^1,2, HAN Jing^2*

1. Department of Ophthalmology, Xi′an Medical University, Xi′an 710068;
2. Department of Ophthalmology, the Second Affiliated Hospital of the Air Force Medical University, Xi′an 710038, China

Received:2023-09-26 Revised:2024-01-24 Online:2024-05-05 Published:2024-04-23
Contact: ^*hanjing.cn@163.com

摘要/Abstract

摘要： 目的探讨褪黑素(MT)对过氧化氢诱导的人脐静脉内皮细胞(HUVECs)氧化应激机制与凋亡标志蛋白质的表达的影响。方法将HUVECs细胞分为空白组和H₂O₂(100、200、300、400、600和700 mmol/L)组,用CCK8法筛选出造模最佳浓度;为了明确MT对H₂O₂造成的氧化应激损伤的影响,将HUVECs细胞分为空白组、H₂O₂组和MT组。Western blot检测细胞中p-p65/p65、SOD2和cleaved-caspase 3的表达;试剂盒检测细胞中超氧化物歧化酶(SOD)活性、丙二醛(MDA)和过氧化氢酶(CAT)浓度;活性氧物质(ROS)染色检测HUVECs细胞内ROS浓度。结果相较于空白组,H₂O₂组细胞内的p-p65/p65和cleaved-caspase 3表达显著上升(P＜0.001),SOD2表达明显下降(P＜0.05);SOD活性(P＜0.001)和CAT浓度(P＜0.01)明显下降,MDA浓度明显上升(P＜0.001);ROS含量显著升高(P＜0.001)。相较于H₂O₂组,MT干预组细胞内的p-p65/p65(P＜0.001)和cleaved-caspase 3表达(P＜0.01)明显下调,SOD2表达明显增高(P＜0.001);并且SOD活性(P＜0.001)和CAT浓度上升(P＜0.05),MDA浓度下降(P＜0.001);从ROS染色的结果看来ROS含量有明显下降(P＜0.001)。结论 MT通过上调SOD2的表达、下调p-p65/p65和cleaved-caspase 3表达,发挥对H₂O₂介导的HUVECs氧化应激损伤的保护作用。

关键词: 生物医学工程, 褪黑素(MT), 分子生物学, 人脐静脉内皮细胞(HUVECs), 氧化应激

Abstract: Objective To investigate the effects of melatonin (MT) on oxidative stress and expression of apoptosis marker protein in human umbilical vein endothelial cells (HUVECs) induced by hydrogen peroxide (H₂O₂). Methods HUVECs were divided into blank group and H₂O₂ (100, 200, 300, 400, 600, 700 mmol/L) groups.CCK8 method was used to select the best concentration for modeling. To clarify the effect of melatonin on oxidative stress injury caused by H₂O₂, HUVECs were divided into control group, H₂O₂ group and melatonin group. Western blot was used to detect the expression of p-p65/p65, SOD2 and cleaved-caspase 3. The activity of super-oxide dismutase (SOD), the concentration of malondialdehyde (MDA) and catalase (CAT) were measred by commercially available kits. Reactive oxygen species (ROS) staining was used to detect ROS content in HUVECs. Results H₂O₂ increased the expression of p-p65/p65 and cleaved-caspase 3(P＜0.001), decreased the expression of SOD2(P＜0.05)its biological activity(P＜0.001) and the concentration of CAT(P＜0.01), elevated the concentration of MDA(P＜0.001) and the level of ROS(P＜0.001)in HUVECs. Compared with H₂O₂ group, MT treatment decreased the expression of p-p65/p65(P＜0.001), SOD2(P＜0.001)and cleaved-caspase 3(P＜0.01), increased the expression of SOD2(P＜0.001)as well as biological activity of SOD(P＜0.001) and the concentration of CAT(P＜0.05) reduced the concentration of MDA(P＜0.001) and the level of ROS(P＜0.001)in HUVECs. Conclusions Melatonin plays a protective role in oxidative stress injury of HUVECs induced by H₂O₂ through up-regulating the expression of SOD2 and down-regulating the expression of p-p65/p65 and cleaved-caspase 3.

Key words: biomedical engineering, melatonin(MT), molecular biology, human umbilical vein endothelial cells (HUVECs), oxidative stress

中图分类号:

杨瀚毅, 宁佳怡, 汪小兰, 张益萌, 谢婷珂, 陈奕玄, 韩静. 褪黑素抑制H₂O₂诱导的人脐静脉内皮细胞氧化应激及凋亡标志蛋白质的表达[J]. 基础医学与临床, 2024, 44(5): 645-650.

YANG Hanyi, NING Jiayi, WANG Xiaolan, ZHANG Yimeng, XIE Tingke, CHEN Yixuan, HAN Jing. Melatonin inhibits H₂O₂-induced oxidative stress and apoptosis marker protein expression in human umbilical vein endothelial cells[J]. Basic & Clinical Medicine, 2024, 44(5): 645-650.

参考文献

[1] Association DBOCM 中国2型糖尿病防治指南(2020年版)(下)[J]. 中国实用内科杂志, 2021, 41: 757-784. doi:10.19538/j.nk2021090106.
[2] Yang J, Liu Z. Mechanistic pathogenesis of endothelial dysfunction in diabetic nephropathy and retinopathy[J]. Front Endocrinol (Lausanne), 2022, 13: 816400.doi:10.3389/fendo.2022.816400.
[3] Kim YM, Kim SJ, Tatsunami R, et al. Ros-induced ros release orchestrated by nox4, nox2, and mitochondria in vegf signaling and angiogenesis[J]. Am J Physiol Cell Physiol, 2017, 312: C749-C764. doi:10.1152/ajpcell.00346.2016.
[4] Sies H. Hydrogen peroxide as a central redox signaling molecule in physiological oxidative stress: oxidative eustress[J]. Redox Biol, 2017, 11: 613-619. doi:10.1016/j.redox.2016.12.035.
[5] Zenin V, Ivanova J, Pugovkina N, et al. Resistance to h(2)o(2)-induced oxidative stress in human cells of different phenotypes[J]. Redox Biol, 2022, 50: 102245. doi:10.1016/j.redox.2022.102245.
[6] Zhao D, Wang H, Chen S, et al. Phytomelatonin: an emerging regulator of plant biotic stress resistance[J]. Trends Plant Sci, 2021, 26: 70-82. doi:10.1016/j.tplants.2020.08.009.
[7] Yan F, Zhao Q, Li Y, et al. The role of oxidative stress in ovarian aging: A review[J]. J Ovarian Res, 2022, 15: 100.doi:10.1186/s13048-022-01032-x.
[8] Zhang X, Zhang D, Li H, et al. Melatonin-mediated suppression of mtROS-JNK-FOXO1 pathway alleviates hypoxia-induced apoptosis in porcine granulosa cells[J]. Antioxidants (Basel), 2023, 12: doi:10.3390/antiox12101881.
[9] Wang J, Hao H, Yao L, et al. Melatonin suppresses migration and invasion via inhibition of oxidative stress pathway in glioma cells[J]. J Pineal Res, 2012, 53: 180-7. doi:10.1111/j.1600-079X.2012.00985.x.
[10] Abdullah SE, Perez-Soler R. Mechanisms of resistance to vascular endothelial growth factor blockade[J]. Cancer, 2012, 118: 3455-67. doi:10.1002/cncr.26540.
[11] Rodríguez ML, Pérez S, Mena-Mollá S, et al. Oxidative stress and microvascular alterations in diabetic retino-pathy: future therapies[J]. Oxid Med Cell Longev, 2019, 2019: 4940825. doi:10.1155/2019/4940825.
[12] Duarte MB, Medeiros BZ, Da Silva Lemos I, et al. Melatonin improves behavioral parameters and oxidative stress in zebrafish submitted to a leucine-induced msud protocol[J]. Metab Brain Dis, 2023, 38: 2105-2114. doi:10.1007/s11011-023-01220-8.
[13] Reiter RJ, Mayo JC, Tan DX, et al. Melatonin as an antioxidant: under promises but over delivers[J]. J Pineal Res, 2016, 61: 253-278. doi:10.1111/jpi.12360.
[14] Nennig SE, Schank JR. The role of NF-κB in drug addiction: beyond inflammation[J]. Alcohol Alcohol, 2017, 52: 172-179. doi:10.1093/alcalc/agw098.

[1]	王瑞, 李蕊, 李运龙, 张玉萍, 于康, 刘燕萍. 锶与妊娠及妊娠期疾病的研究进展[J]. 基础医学与临床, 2024, 44(4): 428-433.
[2]	张威威, 谢婧, 李丽华. 帕立骨化醇抑制氧化应激减轻小鼠肝细胞紧密连接受损[J]. 基础医学与临床, 2024, 44(3): 317-324.
[3]	张秀丽, 胡孝刚, 莫俊, 陈铃. 冠心宁片减轻大鼠颈动脉粥样硬化斑块损伤[J]. 基础医学与临床, 2024, 44(2): 192-198.
[4]	牟杨, 杨志文. 右美托咪定通过激活AMPK减轻TNF-α诱导人成神经细胞瘤细胞系SH-SY5Y损伤[J]. 基础医学与临床, 2024, 44(2): 147-153.
[5]	陈新军, 王卿语, 陈乐, 古春昱, 武卓. 紫檀茋减轻百草枯诱导的模型大鼠肺纤维化[J]. 基础医学与临床, 2023, 43(9): 1383-1389.
[6]	王琼, 董倩兰, 朱燕亭, 金刚, 张琳萍. 桔梗皂苷D减轻大鼠肾缺血/再灌注损伤[J]. 基础医学与临床, 2023, 43(8): 1222-1228.
[7]	杨为祝, 刘忠, 张峰林, 黄延林, 孙瑾, 田新华. NOX4介导的氧化应激参与垂体腺瘤出血性卒中的发生[J]. 基础医学与临床, 2023, 43(7): 1040-1044.
[8]	汪博, 张明焕. CIRC_0044235靶向miR-574-5p减轻IL-1β诱导的人软骨细胞损伤[J]. 基础医学与临床, 2023, 43(3): 419-426.
[9]	张梦瑶, 牛姝, 蔡静, 张立香, 赵志刚. 丹参酮ⅡA减轻高糖诱导的小鼠足细胞系PMC-5凋亡和氧化应激反应[J]. 基础医学与临床, 2023, 43(2): 277-282.
[10]	何綦琪, 吴晃, 李昱卓, 包军胜. 线粒体功能障碍在肾结石成石中作用的研究进展[J]. 基础医学与临床, 2023, 43(2): 306-310.
[11]	袁培根, 陈顺利, 单鸳露, 薛彬彬, 叶玉柱, 林丽娜. 艾司氯胺酮减轻大鼠骨骼肌缺血再灌注损伤[J]. 基础医学与临床, 2023, 43(12): 1822-1826.
[12]	顾炜, 祝剑平, 吴品雯. 丙泊酚减轻低氧诱导的大鼠肾上腺嗜铬细胞瘤细胞系PC12炎性反应和细胞凋亡[J]. 基础医学与临床, 2023, 43(1): 137-143.
[13]	郭丽苹, 苏娟, 田新雁, 陆丽, 胡亚荣, 秦燕. 骨桥蛋白在大鼠肝缺血/再灌注损伤中的作用[J]. 基础医学与临床, 2023, 43(1): 83-86.
[14]	胡博, 阎敬, 冯晨, 王华芹. 国际学生生物化学实验中的云端混合教学模式[J]. 基础医学与临床, 2022, 42(9): 1464-1469.
[15]	余子强, 徐久平, 詹长生, 邵明峰, 邹建安. 输尿管软镜碎石与经皮肾镜碎石对3 cm以下肾结石疗效和炎性反应的影响[J]. 基础医学与临床, 2022, 42(8): 1259-1262.

褪黑素抑制H₂O₂诱导的人脐静脉内皮细胞氧化应激及凋亡标志蛋白质的表达

Melatonin inhibits H₂O₂-induced oxidative stress and apoptosis marker protein expression in human umbilical vein endothelial cells

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价