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[1]白玉杰,王 枭,林 宁,等.甲基胞嘧啶双加氧酶2对低氧人脐静脉内皮细胞损伤的作用研究[J].中华肺部疾病杂志,2025,(03):369-374.[doi:10.3877/cma.j.issn.1674-6902.2025.03.005]
　Bai Yujie,Wang Xiao,Lin Ning,et al.Study on the effects and mechanisms of ten-eleven translocation methylcytosine dioxygenase 2 on hypoxia-induced injury in human umbilical vein endothelial cells[J].,2025,(03):369-374.[doi:10.3877/cma.j.issn.1674-6902.2025.03.005]
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甲基胞嘧啶双加氧酶2对低氧人脐静脉内皮细胞损伤的作用研究(PDF)

《中华肺部疾病杂志》[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2025年03期
页码:
369-374
栏目:
论著
出版日期:
2025-06-25

文章信息/Info

Title:
Study on the effects and mechanisms of ten-eleven translocation methylcytosine dioxygenase 2 on hypoxia-induced injury in human umbilical vein endothelial cells
作者:
白玉杰1; 2; 王 枭3; 林 宁4; 刘佳姣5; 罗书泓6; 冯 健2; 李福祥1; 2
646000 泸州,西南医科大学附属医院呼吸内科1
610083 成都,西部战区总医院重症医学科2
610083 成都,西部战区总医院血液科3
610083 成都,西部战区总医院营养科4
400020 重庆,陆军特色医学中心神经外科5
610299 成都,成都市双流区第一人民医院呼吸内科6
Author(s):
Bai Yujie1; 2;  Wang Xiao3;  Lin Ning4;  Liu Jiajiao5;  Luo Shuhong6;  Feng Jian2;  Li Fuxiang1; 2.
1Department of Respiratory Medicine, Affiliated Hospital of Southwest Medical University, Luzhou 646000, China; 2Department of Critical Care Medicine. The General Hospital of Western Theater Command, Chengdu 610083, China; 3Department of Hematology, The General Hospital of Western Theater Command, Chengdu 610083, China; 4Department of Nutrition, The General Hospital of Western Theater Command, Chengdu 610083, China; 5Neurosurgery, Army Characteristic Medical Center, Chongqing 400020, China; 6Department of Respiratory Medicine, The First People's Hospital of Shuangliu District, Chengdu 610299, China.
关键词:
甲基胞嘧啶双加氧酶2;  氧化应激;  内皮细胞损伤;  静脉血栓栓塞;  高原低氧
Keywords:
Ten-eleven translocation methylcytosine dioxygenase 2;  Oxidative stress;  Endothelial cell injury;  Venous thromboembolism;  High altitude hypoxia
分类号:
R563
DOI:
10.3877/cma.j.issn.1674-6902.2025.03.005
摘要:
目的 分析甲基胞嘧啶双加氧酶2(ten-eleven translocation methylcytosine dioxygenase 2, TET2)对低氧人脐静脉内皮细胞(Human umbilical vein endothelial cells, HUVECs)损伤的作用及机制。方法 采用生理盐水+常氧处理HUVECs为Norm-C组,凝血酶+常氧处理为Norm-M组,生理盐水+低氧处理为Hypo-M组,低氧+凝血酶处理为Hypo-M组; 低氧+凝血酶+空载体处理为空载体组、低氧+凝血酶+携带TET2重组腺病毒处理为AdTET2组; 低氧+凝血酶+H2O2处理为H2O2组,低氧+凝血酶+携带TET2基因重组腺病毒+H2O2处理为AdTET2+H2O2组。观察细胞超微结构,监测组织型纤溶酶原激活物(tissue plasminogen activator, t-PA)、血栓调节蛋白(thrombomodulin, TM)、血管内皮生长因子(vascular endothelial growth factor, VEGF)、血管细胞黏附分子(vascular cell adhesion molecule, VCAM)、血管性血友病因子(von willebrand factor, vWF)、纤溶酶原激活物抑制剂-1(plasminogen activator inhibitor-1, PAI-1)和活性氧(reactive oxygen species, ROS)、超氧化物歧化酶(superoxide dismutase, SOD)、丙二醛(malondialdehyde, MDA)。结果 Hypo-C组较Norm-C组胞浆内细胞器肿胀和空泡化,细胞膜完整,Norm-M组和Hypo-M组细胞膜破裂和细胞质渗漏。Hypo-M组t-PA(2.03±0.04)ng/ml、TM(1.04±0.06)ng/ml、SOD活性(12.97±1.79)U/mg、TET2 mRNA(0.45±0.04)低于Norm-C组t-PA(3.29±0.25)ng/ml、TM(1.45±0.11)ng/ml、SOD活性(19.66±2.15)U/mg、TET2 mRNA(1.00±0.07); Hypo-M组VEGF(435.03±18.27)pg/ml、VCAM(74.14±4.47)ng/ml、vWF(87.22±6.57)ng/ml、PAI-1(12.32±0.28)pg/ml、ROS(81 911.90±3 011.78)、MDA(0.87±0.10)nmol/mg高于Norm-C组VEGF(328.55±21.02)pg/ml、VCAM(56.74±3.95)ng/ml、vWF(66.27±4.91)ng/ml、PAI-1(8.07±0.43)pg/ml、ROS(46 424.60±2 040.93)、MDA(0.55±0.06)nmol/mg。AdTET2组与模型组或空载体组相比,t-PA、TM、SOD升高,VEGF、VCAM、vWF、PAI-1、ROS、MDA降低。AdTET2+H2O2组TM、t-PA高于H2O2组,VEGF、VCAM、vWF、PAI-1、ROS、MDA低于H2O2组。结论 低氧和凝血酶诱导HUVECs损伤,TET2过表达减轻氧化应激,发挥保护作用。
Abstract:
Objective To investigate the effects and mechanisms of ten-eleven translocation methylcytosine dioxygenase 2(TET2)on hypoxia-induced injury in human umbilical vein endothelial cells(HUVECs). Methods Human umbilical vein endothelial cells(HUVECs)were treated as follows: HUVECs treated with normal saline under normoxia were defined as the Norm-C group; HUVECs treated with thrombin under normoxia were defined as the Norm-M group; HUVECs treated with normal saline under hypoxia were defined as the Hypo-M group; HUVECs treated with hypoxia and thrombin were also defined as the Hypo-M group. Additionally, HUVECs treated with hypoxia, thrombin, and an adenovirus empty vector without TET2 were defined as the empty vector group; HUVECs treated with hypoxia, thrombin, and a recombinant adenovirus carrying the TET2 gene were defined as the AdTET2 group; HUVECs treated with hypoxia, thrombin, and H2O2 were defined as the H2O2 group; HUVECs treated with hypoxia, thrombin, a recombinant adenovirus carrying the TET2 gene, and H2O2 were defined as the AdTET2+H2O2 group. The ultrastructure of the cells was observed, and the levels of tissue plasminogen activator(t-PA), thrombomodulin(TM), vascular endothelial growth factor(VEGF), vascular cell adhesion molecule(VCAM), von Willebrand factor(vWF), plasminogen activator inhibitor-1(PAI-1), reactive oxygen species(ROS), superoxide dismutase(SOD), and malondialdehyde(MDA)were monitored. Results Compared with the Norm-C group, the Hypo-C group exhibited swollen and vacuolated organelles in the cytoplasm, but the cell membranes remained intact. In contrast, the Norm-M and Hypo-M groups showed ruptured cell membranes and cytoplasmic leakage. In the Hypo-M group, the levels of t-PA(2.03±0.04)ng/ml, TM(1.04±0.06)ng/ml, SOD activity(12.97±1.79)U/mg, and TET2 mRNA(0.45±0.04)were lower than those in the Norm-C group t-PA(3.29±0.25)ng/ml, TM(1.45±0.11)ng/ml, SOD activity(19.66±2.15)U/mg, TET2 mRNA(1.00±0.07). Conversely, the levels of VEGF(435.03±18.27)pg/ml, VCAM(74.14±4.47)ng/ml, vWF(87.22±6.57)ng/ml, PAI-1(12.32±0.28)pg/ml, ROS(81 911.90±3 011.78), and MDA(0.87±0.10)nmol/mg were higher in the Hypo-M group than in the Norm-C group VEGF(328.55±21.02)pg/ml, VCAM(56.74±3.95)ng/ml, vWF(66.27±4.91)ng/ml, PAI-1(8.07±0.43)pg/ml, ROS(46 424.60±2 040.93), MDA(0.55±0.06)nmol/mg. Compared with the model group or the empty vector group, the AdTET2 group showed increased levels of t-PA, TM, and SOD activity, while VEGF, VCAM, vWF, PAI-1, ROS, and MDA were decreased. In the AdTET2+H2O2 group, the levels of TM and t-PA were higher than those in the H2O2 group, while VEGF, VCAM, vWF, PAI-1, ROS, and MDA were lower than those in the H2O2 group. Conclusion Hypoxia and thrombin induce injury in HUVECs, while overexpression of TET2 alleviates oxidative stress and exerts a protective effect.

参考文献/References:

1 Imiela AM, Mikoajczyk TP, Guzik TJ, et al. Acute pulmonary embolism and immunity in animal models[J]. Arch Immunol Ther Exp(Warsz), 2024, 72(1): doi: 10.2478/aite-2024-0003.
2 Xia YQ, Tang L, Hu Y. Advances in the genetics of venous thromboembolic disease[J]. Zhonghua Xue Ye Xue Za Zhi, 2024, 45(12): 1144-1147.
3 Simonneau G, Dorfmüller P, Guignabert C, et al. Chronic thromboembolic pulmonary hypertension: the magic of pathophysiology[J]. Ann Cardiothorac Surg, 2022, 11(2): 106-119.
4 Zhou C, Zhou Y, Ma W, et al. Revisiting Virchow's triad: exploring the cellular and molecular alterations in cerebral venous congestion[J]. Cell Biosci, 2024, 14(1): 131.
5 Guo J, Wang Z, Wu J, et al. Endothelial SIRT6 is vital to prevent hypertension and associated cardiorenal injury through targeting Nkx3.2-GATA5 signaling[J]. Circ Res, 2019, 124(10): 1448-1461.
6 Ashry NA, Abdelaziz RR, Suddek GM. The potential effect of imatinib against hypercholesterolemia induced atherosclerosis, endothelial dysfunction and hepatic injury in rabbits[J]. Life Sci, 2020, 243: 117275.
7 Chen Q, Li X, Kong L, et al. miR-101-3p induces vascular endothelial cell dysfunction by targeting tet methylcytosine dioxygenase 2[J]. Acta Biochim Biophys Sin(Shanghai), 2020, 52(2): 180-191.
8 Schönichen C, Sun S, Middelveld H, et al. Functionally distinct anticoagulant mechanisms of endothelial cells[J]. Thromb Res, 2024, 244: 109208.
9 Ninivaggi M, Swieringa F, Middelveld H, et al. Exercise and hypoxia-induced hypercoagulability is counterbalanced in women in part by decreased platelet reactivity[J]. Thromb Res, 2024, 234: 142-150.
10 Vijay A, Jha PK, Parveen S, et al. Aberrant promoter hypermethylation regulates thrombomodulin in high altitude induced deep vein thrombosis[J]. Thromb Res, 2022, 215: 5-13.
11 Tourn J, Crescence L, Bruzzese L, et al. Cellular and molecular mechanisms leading to air travel-induced thrombosis[J]. Circ Res, 2025, 136(1): 115-134.
12 Shaydakov M E, Diaz J A, Eklöf B, et al. Venous valve hypoxia as a possible mechanism of deep vein thrombosis: a scoping review[J]. Int Angiol, 2024, 43(3): 309-322.
13 Liu X, Li Z. The role and mechanism of epigenetics in anticancer drug-induced cardiotoxicity[J]. Basic Res Cardiol, 2025, 120(1): 11-24.
14 Li X, Dong X, Lu W, et al. Integrated analysis of gene expression and methylation data to identify potential biomarkers related to atherosclerosis onset[J]. Oxid Med Cell Longev, 2022, 2022: 5493051.
15 Kumar A, Misra S, Nair P, et al. Epigenetics mechanisms in ischemic stroke: A promising avenue?[J]. J Stroke Cerebrovasc Dis, 2021, 30(5): 105690.
16 He J, Lin M, Zhang X, et al. TET2 is required to suppress mTORC1 signaling through urea cycle with therapeutic potential[J]. Cell Discov, 2023, 9(1): 84.
17 Okashita N, Kuroki S, Maeda R, et al. TET2 catalyzes active DNA demethylation of the Sry promoter and enhances its expression[J]. Sci Rep, 2019, 9(1): 13462.
18 Wang Z, Liu W, Wang D, et al. TET2 mutation may be more valuable in predicting thrombosis in ET patients compared to PV patients: A preliminary report[J]. J Clin Med, 2022, 11(22): 6615.
19 Wang J, Uryga AK, Reinhold J, et al. Vascular smooth muscle cell senescence promotes atherosclerosis and features of plaque vulnerability[J]. Circulation, 2015, 132(20): 1909-1919.
20 Zeng J, Tao J, Xia L, et al. Melatonin inhibits vascular endothelial cell pyroptosis by improving mitochondrial function via up-regulation and demethylation of UQCRC1[J]. Biochem Cell Biol, 2021, 99(3): 339-347.
21 Shi Y, Li B, Huang X, et al. Loss of TET2 impairs endothelial angiogenesis via downregulating STAT3 target genes[J]. Cell Biosci, 2023, 13(1): 12.
22 Yang X, Xiang Y, Wang F, et al. Expressions and relationship of Krüppel-like factor 15 and endothelial nitric oxide synthase in experimental deep venous thrombosis[J]. Ann Transl Med, 2020, 8(17): 1090.
23 Qiao X, Cao S, Chen S, et al. Salvianolic acid A alleviates H(2)O(2)-induced endothelial oxidative injury via miR-204-5p[J]. Sci Rep, 2024, 14(1): 11931.
24 Peng J, Tang Z, Ren Z, et al. TET2 Protects against oxLDL-Induced HUVEC Dysfunction by Upregulating the CSE/H(2)S System[J]. Front Pharmacol, 2017, 8: 486.
25 Iba T, Helms J, Levi M, et al. Thromboinflammation in acute injury: infections, heatstroke, and trauma[J]. J Thromb Haemost, 2024, 22(1): 7-22.
26 Srivastava S, Kumari B, Garg I, et al. Targeted gene expression study using TaqMan low density array to gain insights into venous thrombo-embolism(VTE)pathogenesis at high altitude[J]. Blood Cells Mol Dis, 2020, 82: 102421.
27 He JZ, Ho JJD, Gingerich S, et al. Enhanced translation of heme oxygenase-2 preserves human endothelial cell viability during hypoxia[J]. J Biol Chem, 2010, 285(13): 9452-9461.
28 Incalza MA, D'Oria R, Natalicchio A, et al. Oxidative stress and reactive oxygen species in endothelial dysfunction associated with cardiovascular and metabolic diseases[J]. Vascul Pharmacol, 2018, 100: 1-19.
29 Sum H, Brewer AC. Epigenetic modifications as therapeutic targets in atherosclerosis: a focus on DNA methylation and non-coding RNAs[J]. Front Cardiovasc Med, 2023, 10: 1183181.
30 Li A, Tan L, Zhang S, et al. Low shear stress-induced endothelial mesenchymal transformation via the down-regulation of TET2[J]. Biochem Biophys Res Commun, 2021, 545: 20-26.
31 Yu Y, Yan R, Chen X, et al. Paeonol suppresses the effect of ox-LDL on mice vascular endothelial cells by regulating miR-338-3p/TET2 axis in atherosclerosis[J]. Mol Cell Biochem, 2020, 475(1-2): 127-135.
32 Rauch PJ, Gopakumar J, Silver AJ, et al. Loss-of-function mutations in Dnmt3a and Tet2 lead to accelerated atherosclerosis and concordant macrophage phenotypes[J]. Nat Cardiovasc Res, 2023, 2(9): 805-818.
33 Gao Q, Shen K, Xiao M. TET2 mutation in acute myeloid leukemia:biology, clinical significance, and therapeutic insights[J]. Clin Epigenetics, 2024, 16(1): 155.
34 Wen H, Liu X, Zhu Z, et al. TET2 is downregulated in early esophageal squamous cell carcinoma and promotes esophageal squamous cell malignant behaviors[J]. Dig Dis Sci, 2024, 69(7): 2462-2476.
35 Hu F, Chen X, Gao J, et al. CircDIP2C ameliorates oxidized low-density lipoprotein-induced cell dysfunction by binding to miR-556-5p to induce TET2 in human umbilical vein endothelial cells[J]. Vascul Pharmacol, 2021, 139: 106887.

备注/Memo

备注/Memo:
基金项目: 西部战区总医院应用基础研究项目重点项目(2021-XZYG-A08)
四川省干部保健科研课题(川干研2022-1303)
通信作者: 李福祥, Email: lfx98@163.com
冯 健, Email: ironvon@foxmail.com
更新日期/Last Update: 2025-06-25