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

doi:10.3877/cma.j.issn.1674-6902.2025.03.005

中华肺部疾病杂志(电子版) ›› 2025, Vol. 18 ›› Issue (03) : 369 -374. doi: 10.3877/cma.j.issn.1674-6902.2025.03.005

论著

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

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^,^†(

)

^1. 646000 泸州，西南医科大学附属医院呼吸内科
^2. 610083 成都，西部战区总医院重症医学科
^3. 610083 成都，西部战区总医院血液科
^4. 610083 成都，西部战区总医院营养科
^5. 400020 重庆，陆军特色医学中心神经外科
^6. 610299 成都，成都市双流区第一人民医院呼吸内科

收稿日期:2025-03-26 出版日期:2025-06-25
通信作者: 冯健, 李福祥
基金资助:
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）

Study on the effects and mechanisms of ten-eleven translocation methylcytosine dioxygenase 2 on hypoxia-induced injury in human umbilical vein endothelial cells

Yujie Bai¹^,², Xiao Wang³, Ning Lin⁴, Jiajiao Liu⁵, Shuhong Luo⁶, Jian Feng²^,^†(), Fuxiang Li¹^,²^,^†()

^1. Department of Respiratory Medicine，Affiliated Hospital of Southwest Medical University，Luzhou 646000，China
^2. Department of Critical Care Medicine. The General Hospital of Western Theater Command，Chengdu 610083，China
^3. Department of Hematology，The General Hospital of Western Theater Command，Chengdu 610083，China
^4. Department of Nutrition，The General Hospital of Western Theater Command，Chengdu 610083，China
^5. Neurosurgery，Army Characteristic Medical Center，Chongqing 400020，China
^6. Department of Respiratory Medicine，The First People's Hospital of Shuangliu District，Chengdu 610299，China

Received:2025-03-26 Published:2025-06-25
Corresponding author: Jian Feng, Fuxiang Li

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引用本文：

白玉杰, 王枭, 林宁, 刘佳姣, 罗书泓, 冯健, 李福祥. 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/OL]. 中华肺部疾病杂志(电子版), 2025, 18(03): 369-374.

Yujie Bai, Xiao Wang, Ning Lin, Jiajiao Liu, Shuhong Luo, Jian Feng, Fuxiang Li. Study on the effects and mechanisms of ten-eleven translocation methylcytosine dioxygenase 2 on hypoxia-induced injury in human umbilical vein endothelial cells[J/OL]. Chinese Journal of Lung Diseases(Electronic Edition), 2025, 18(03): 369-374.

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。

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

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.

Key words: Ten-eleven translocation methylcytosine dioxygenase 2, Oxidative stress, Endothelial cell injury, Venous thromboembolism, High altitude hypoxia

表1 每组HUVECs 细胞因子及氧化应激标志物

监测指标	模型组	空载体组	AdTET2 组	P 值
TM[ng/ml,(x̄ ± s)]	1.07±0.06	1.11±0.02	1.42±0.12	0.002
t-PA[ng/ml,(x̄ ± s)]	1.97±0.12	2.03±0.11	3.29±0.24	0.000
PAI-1[ng/ml,(x̄ ± s)]	12.32±0.28	12.99±0.71	8.47±0.63	0.000
VCAM[ng/ml,(x̄ ± s)]	74.61±4.94	75.77±5.50	57.40±5.89	0.008
vWF[ng/ml,(x̄ ± s)]	94.22±9.26	87.70±6.60	67.27±7.89	0.006
VEGF[ng/ml,(x̄ ± s)]	435.04±18.28	450.62±25.27	342.55±40.69	0.009
ROS(x̄ ± s)	81 380.87±2 031.72	82 379.03±2 060.31	42 695.33±2 261.16	0.000
MDA[nmol/mg,(x̄ ± s)]	0.81±0.05	0.84±0.06	0.62±0.03	0.003
SOD(x̄ ± s)	13.35±0.59	13.57±0.02	20.17±1.21	0.000

表1 每组HUVECs 细胞因子及氧化应激标志物

监测指标	模型组	空载体组	AdTET2 组	P 值
TM[ng/ml,(x̄ ± s)]	1.07±0.06	1.11±0.02	1.42±0.12	0.002
t-PA[ng/ml,(x̄ ± s)]	1.97±0.12	2.03±0.11	3.29±0.24	0.000
PAI-1[ng/ml,(x̄ ± s)]	12.32±0.28	12.99±0.71	8.47±0.63	0.000
VCAM[ng/ml,(x̄ ± s)]	74.61±4.94	75.77±5.50	57.40±5.89	0.008
vWF[ng/ml,(x̄ ± s)]	94.22±9.26	87.70±6.60	67.27±7.89	0.006
VEGF[ng/ml,(x̄ ± s)]	435.04±18.28	450.62±25.27	342.55±40.69	0.009
ROS(x̄ ± s)	81 380.87±2 031.72	82 379.03±2 060.31	42 695.33±2 261.16	0.000
MDA[nmol/mg,(x̄ ± s)]	0.81±0.05	0.84±0.06	0.62±0.03	0.003
SOD(x̄ ± s)	13.35±0.59	13.57±0.02	20.17±1.21	0.000

图1 HUVECs 中TM、t-PA、PAI-1、VCAM、vWF、VEGF 水平。图A 为细胞上清液TM 浓度；图B 为细胞上清液t-PA 浓度；图C为细胞上清液PAI-1 浓度；图D 为细胞上清液VCAM 浓度；图E 为细胞上清液vWF 浓度；图F 为细胞上清液VEGF 浓度。与模型组相比：*P＜0.05，**P＜0.01，***P＜0.001；与AdTET2 组相比：&P＜0.05，&&P＜0.01，&&&P＜0.001；与H₂O₂ 组相比：#P＜0.05，##P＜0.01，###P＜0.001

1	Imiela AM，Mikołajczyk 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 hypoxiainduced 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 oxLDLInduced 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 lowdensity 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.

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Study on the effects and mechanisms of ten-eleven translocation methylcytosine dioxygenase 2 on hypoxia-induced injury in human umbilical vein endothelial cells

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Study on the effects and mechanisms of ten-eleven translocation methylcytosine dioxygenase 2 on hypoxia-induced injury in human umbilical vein endothelial cells