硫化氢对低氧性肺动脉高压大鼠FDX1-铜死亡途径的干预

doi:10.12092/j.issn.1009-2501.2026.07.002

中国临床药理学与治疗学 ›› 2026, Vol. 31 ›› Issue (7): 875-884.doi: 10.12092/j.issn.1009-2501.2026.07.002

硫化氢对低氧性肺动脉高压大鼠FDX1-铜死亡途径的干预

王淑远¹(), 黄曼²(), 陈思安², 韩丛美³, 程缘², 袁琳波³, 叶环¹^,*(), 王万铁²^,*()

1. 温州市人民医院呼吸与危重症医学科，温州 325400，浙江
2. 温州医科大学基础医学院病理学与病理生理学系，温州 325035，浙江
3. 温州医科大学基础医学院机能学系，温州 325035，浙江

收稿日期:2025-12-04 修回日期:2026-03-27 出版日期:2026-07-26 发布日期:2026-08-04
通讯作者: 叶环,王万铁 E-mail:944372370@qq.com;1440480267@qq.com;wsyyehuan@126.com;wwt@wmu.edu.cn
作者简介:王淑远，女，主治医师，研究方向：肺动脉高压及缺血再灌注损伤。E-mail：944372370@qq.com|黄曼，共同第一作者，女，硕士研究生，研究方向：肺动脉高压。E-mail：1440480267@qq.com
基金资助:
浙江省介入肺脏病重点实验室建设项目（2019E10014）

Intervention of hydrogen sulfide on FDX1-cuproptosis pathway in rats with hypoxemic pulmonary hypertension

Shuyuan WANG¹(), Man HUANG²(), Sian CHEN², Congmei HAN³, Yuan CHENG², Linbo YUAN³, Huan YE¹^,*(), Wantie WANG²^,*()

1. Department of Respiratory and Critical Care Medicine, Wenzhou People's Hospital, Wenzhou 325400, Zhejiang, China
2. Department of Pathology and Pathophysiology, School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou 325035, Zhejiang, China
3. Department of Functional Medicine, School of Basic Medicine, Wenzhou Medical University, Wenzhou 325035, Zhejiang, China

Received:2025-12-04 Revised:2026-03-27 Online:2026-07-26 Published:2026-08-04
Contact: Huan YE,Wantie WANG E-mail:944372370@qq.com;1440480267@qq.com;wsyyehuan@126.com;wwt@wmu.edu.cn

摘要/Abstract

摘要：

目的: 探究硫化氢（hydrogen sulfide，H₂S）是否通过铁氧化还原蛋白1（ferredoxin 1，FDX1）-铜死亡途径来改善大鼠低氧性肺动脉高压（hypoxic pulmonary hypertension，HPH）。方法: 雄性Sprague Dawley（SD）大鼠随机分为Control组、Hypoxia组和Hypoxia+NaHS组，每组各10只。Control组大鼠置于常氧（21% O₂）环境中，每天腹腔注射等体积生理盐水；Hypoxia组大鼠和Hypoxia+NaHS组大鼠置于低氧箱中，每天腹腔注射14 μmol/kg的NaHS。造模21 d结束后，用右心导管法检测各组大鼠的平均肺动脉压（mPAP），称重并计算右心室肥厚指数（RVHI）；ELISA法检测内皮素-1（ET-1）、一氧化氮（NO）和前列环素（PGI₂）的含量；电感耦合等离子体质谱（ICP-MS）法检测铜离子浓度；Western blot法检测铜转运蛋白1（CTR1）、铜转运ATP酶α（ATP7A）、铜转运ATP酶β（ATP7B）、FDX1、硫辛酸合成酶（LIAS）、二氢硫辛酸转乙酰基酶（DLAT）、二氢硫辛酸转乙酰基酶二聚体（DLAT Dimer）、二氢硫辛酰胺S-琥珀酰转移酶（DLST）；RT-qPCR法检测FDX1、LIAS、DLAT、二氢硫辛酰胺脱氢酶（DLD）mRNA表达水平；JC-1试剂盒检测线粒体膜电位变化；化学发光法检测ATP含量；ELISA法检测丙酮酸脱氢酶E1亚基α1（PDHA1）含量；磺基水杨酸法检测柠檬酸（CA）含量。结果: 与Control组比较，Hypoxia组中mPAP和RVHI显著上升；ET-1的浓度显著升高而NO和PGI₂的浓度显著降低；铜离子浓度显著升高；铜转运蛋白1表达含量显著升高，铜外排蛋白ATP7A、ATP7B表达含量显著降低；铜死亡相关蛋白FDX1、LIAS、DLAT、DLST表达含量均显著降低而DLAT二聚体蛋白表达含量显著升高；FDX1、LIAS、DLAT、DLD基因mRNA表达水平显著降低；线粒体功能指标JC-1和ATP表达水平显著降低；三羧酸循环指标PDHA1和CA表达含量显著降低（均P<0.05）。与Hypoxia组比较，Hypoxia+NaHS组中的上述指标的变化均可被治疗药物NaHS显著逆转。结论: H₂S可能通过调控FDX1-铜死亡途径影响肺组织细胞线粒体功能、代谢，从而缓解肺血管收缩，改善肺动脉高压。

关键词: 铜离子, 肺动脉高压, 铜死亡, 硫化氢

Abstract:

AIM: To investigate whether hydrogen sulfide (H₂S) improves hypoxic pulmonary hypertension (HPH) in rats through the FDX1-cuproptosis pathway. METHODS: Male Sprague-Dawley (SD) rats were randomly assigned to the Control group; Hypoxia group and Hypoxia+NaHS group, with 10 rats in each group. Rats in the Control group were housed in a normoxic environment (21% O₂) and received a daily intraperitoneal injection of an equal volume of normal saline; rats in the Hypoxia group and Hypoxia+NaHS group were placed in hypoxic chambers and received a daily intraperitoneal injection of 14 μmol/kg NaHS. After 21 days of model establishment, the mean pulmonary arterial pressure (mPAP) of each group was measured by right heart catheterization and the right ventricular hypertrophy index (RVHI) was calculated after weighing. The levels of endothelin-1 (ET-1), nitric oxide (NO) and prostacyclin (PGI₂) were measured by ELISA. Copper ion concentration was detected by inductively coupled plasma mass spectrometry (ICP-MS). The protein levels of copper transporter 1 (CTR1), copper-transporting ATPase α (ATP7A), copper-transporting ATPase β (ATP7B), ferredoxin 1 (FDX1), lipoic acid synthase (LIAS), dihydrolipoamide S-acetyltransferase (DLAT), dihydrolipoamide S-acetyltransferase dimer (DLAT Dimer) and dihydrolipoamide S-succinyltransferase (DLST) were detected by Western blot. The mRNA expression levels of FDX1, LIAS, DLAT and dihydrolipoamide dehydrogenase (DLD) were detected by RT-qPCR. Mitochondrial membrane potential was assessed using the JC-1 kit. ATP content was measured by chemiluminescence. The level of pyruvate dehydrogenase E1 subunit alpha 1 (PDHA1) was measured by ELISA. Citric acid (CA) content was measured by the sulfosalicylic acid method. RESULTS: Compared with the Control group, the Hypoxia group showed significantly increased mPAP and RVHI; significantly elevated ET-1 concentration while NO and PGI₂ concentrations were significantly decreased; significantly increased copper ion concentration; significantly elevated expression of copper transporter 1 while the expressions of copper efflux proteins ATP7A and ATP7B were significantly decreased; the expressions of cuproptosis-related proteins FDX1, LIAS, DLAT and DLST were all significantly decreased, whereas the expression of DLAT dimer is significantly increased; the mRNA expression levels of FDX1, LIAS, DLAT and DLD were significantly decreased; the mitochondrial function indicators JC-1 and ATP levels were significantly decreased and the tricarboxylic acid cycle indicators PDHA1 and CA levels were significantly decreased (all P<0.05). Compared with the Hypoxia group, all the above changes in the Hypoxia+NaHS group were significantly reversed by the therapeutic agent NaHS. CONCLUSION: H₂S may regulate the FDX1-cuproptosis pathway, thereby affecting mitochondrial function and metabolism in lung tissue cells, alleviating pulmonary vasoconstriction and ameliorating pulmonary hypertension.

Key words: copper ion, pulmonary arterial hypertension, cuproptosis, hydrogen sulfide

中图分类号:

王淑远, 黄曼, 陈思安, 韩丛美, 程缘, 袁琳波, 叶环, 王万铁. 硫化氢对低氧性肺动脉高压大鼠FDX1-铜死亡途径的干预[J]. 中国临床药理学与治疗学, 2026, 31(7): 875-884.

Shuyuan WANG, Man HUANG, Sian CHEN, Congmei HAN, Yuan CHENG, Linbo YUAN, Huan YE, Wantie WANG. Intervention of hydrogen sulfide on FDX1-cuproptosis pathway in rats with hypoxemic pulmonary hypertension[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2026, 31(7): 875-884.

图/表 8

表 1

Table 1 Sequences of the primers for RT-qPCR

Name	Primer sequence (5'-3')
β-actin	Forward: GAGAGGGAAATCGTGCGTA
	Reverse: GGAGGAAGAGGATGCGG
FDX1	Forward: CAGTCCACTTCAAGAACCGAGATG
	Reverse: GATGACAGGTAGAGCAAGCCAAAG
LIAS	Forward: ACAGTGTCATGCTTGAAGGAAAGG
	Reverse: ATTCTGGAACCGTCTCTACATTGTG
DLAT	Forward: GGTGTTCGTTAGCCCTCTTGC
	Reverse: TGTCCTTCTTGATGATTCTGCCTTC
DLD	Forward: CCAGGTGCTGGAGAAATGGT
	Reverse: TGGTTTGCCAAAAGACGCAG

图 1

Fig.1 Effect of H2S on hemodynamic indexes in hypoxia pulmonary hypertension (HPH) rats ($ \overline{x} $±s, n=6) A: mean pulmonary artery pressure; B: mean pulmonary artery pressure waveform diagram; C: right ventricular hypertrophy index. cP<0.01, compared with control group; fP<0.01, compared with Hypoxia group.

图 2

Fig.2 Effect of H2S on ET-1, NO and PGI2 levels in the lung tissues of rats in each group ($ \overline{x} $±s, n=5) A: statistical chart of ET-1 concentration; B: statistical chart of NO concentration; C: statistical chart of PGI2 concentration. cP<0.01, compared with control group; eP<0.05, fP<0.01, compared with Hypoxia group.

图 3

Fig.3 Effect of H2S on copper ions in lung tissues of rats in different groups and on the expression of copper transport proteins ($ \overline{x} $±s, n=6) A: copper ion content in lung tissue of rats; B: protein expression levels of copper transporters proteins in lung tissue of rats; C: copper transporters proteins Western blot statistical chart. bP<0.05, cP<0.01, compared with control group; eP<0.05, fP<0.01, compared with Hypoxia group.

图 4

Fig.4 Effect of H2S on the expression of cuproptosis related proteins in the lung tissues of rats in each group（$ \overline{x} $±s，n=6) A: protein expression levels of cuproptosis related proteins in lung tissue of rats; B: cuproptosis related proteins Western blot statistical chart. cP<0.01, compared with control group; eP<0.05, fP<0.01, compared with Hypoxia group.

图 5

Fig.5 mRNA levels of FDX1, LIAS, DLAT and DLD in lung tissues of each group ($ \overline{x} $±s, n=6) A: relative mRNA level of FDX1; B: relative mRNA level of LIAS; C: relative mRNA level of DLAT; D: relative mRNA level of DLD. cP<0.01, compared with control group; eP<0.05, fP<0.01, compared with Hypoxia group.

图 6

Fig.6 Expression levels of JC-1 and ATP content in lung tissues of eachgroup ($ \overline{x} $±s, n=5) A: relative JC-1 level of each group; B: ATP concentration in each group. cP<0.01, compared with control group; fP<0.01, compared with Hypoxia group.

图 7

Fig.7 Expression levels of PDHA1 and CA concentrations in lung tissues of each group ($ \overline{x} $±s,n=5) A: expression level of PDHA1; B: expression level of CA. cP<0.01, comparedwith control group; eP<0.05, fP<0.01, compared with Hypoxia group.

参考文献 28

1	李彦霖, 王一川, 吴剑华, 等. 伴肺动脉高压的系统性红斑狼疮患者肝损伤1例报告[J]. 临床肝胆病杂志, 2022, 38 (2): 365- 368. doi: 10.3969/j.issn.1001-5256.2022.02.031
2	Nakamura K, Akagi S, Ejiri K, et al. Pathophysiology of group 3 pulmonary hypertension associated with lung diseases and/or hypoxia[J]. Int J Mol Sci, 2025, 26 (2): 835. doi: 10.3390/ijms26020835
3	Bisserier M, Salami A, Zalavadia S, et al. Hypoxia-induced pulmonary hypertension in adults and newborns: implications for drug development[J]. Drug Discov Today, 2024, 29 (6): 103978. doi: 10.1016/j.drudis.2024.104015
4	Bogaard HJ, Mizuno S, Guignabert C, et al. Copper dependence of angioproliferation in pulmonary arterial hypertension in rats and humans[J]. Am J Respir Cell Mol Biol, 2012, 46 (5): 582- 591. doi: 10.1165/rcmb.2011-0296OC
5	Tsvetkov P, Coy S, Petrova B, et al. Copper induces cell death by targeting lipoylated TCA cycle proteins[J]. Science, 2022, 375 (6586): 1254- 1261. doi: 10.1126/science.abf0529
6	Cirino G, Szabo C, Papapetropoulos A. Physiological roles of hydrogen sulfide in mammalian cells, tissues, and organs[J]. Physiol Rev, 2023, 103 (1): 31- 276. doi: 10.1152/physrev.00028.2021
7	Sun HJ, Lu Y, Zhu Y, et al. Pharmacology of hydrogen sulfide and its donors in cardiometabolic diseases[J]. Pharmacol Rev, 2024, 76 (5): 846- 895. doi: 10.1124/pharmrev.123.000928
8	Cobine PA, Brady DC. Cuproptosis: cellular and molecular mechanisms underlying copper-induced cell death[J]. Mol Cell, 2022, 82 (10): 1786- 1787. doi: 10.1016/j.molcel.2022.05.001
9	Hanna DA, Chen B, Shah YM, et al. H₂S remodels mitochondrial ultrastructure and destabilizes respiratory supercomplexes[J]. J Biol Chem, 2025, 301 (5): 108433. doi: 10.1016/j.jbc.2025.108433
10	Nomani L, Zehra A, Choudhary S, et al. Exogenous hydrogen sulphide alleviates copper stress impacts in Artemisia annua L: growth, antioxidant metabolism, glandular trichome development and artemisinin biosynthesis[J]. Plant Biol, 2022, 24 (4): 642- 651. doi: 10.1111/plb.13242
11	Rai P, Singh VP, Peralta-Videa J, et al. Hydrogen sulfide (H₂S) underpins the beneficial silicon effects against the copper oxide nanoparticles (CuO NPs) phytotoxicity in Oryza sativa seedlings[J]. J Hazard Mater, 2021, 415, 124907. doi: 10.1016/j.jhazmat.2020.124907
12	程缘, 田云娜, 黄曼, 等. 硫化氢通过抑制有氧糖酵解-细胞焦亡改善大鼠低氧性肺动脉高压[J]. 生理学报, 2025, 77 (3): 465- 471. doi: 10.13294/j.aps.2025.0015
13	刘军杰, 黄国雄, 黎静, 等. 硫化氢对高肺血流性肺动脉高压大鼠平滑肌细胞线粒体损伤的保护作用及其机制[J]. 吉林大学学报(医学版), 2014, 40 (3): 593- 597. doi: 10.13481/j.1671-587x.20140326
14	Zafeiropoulos S, Ahmed U, Bekiaridou A, et al. Ultrasound neuromodulation of an anti-inflammatory pathway at the spleen improves experimental pulmonary hypertension[J]. Circ Res, 2024, 135 (1): 41- 56. doi: 10.1161/CIRCRESAHA.123.323679
15	罗勤. 中国肺动脉高压诊治临床路径[J]. 中国循环杂志, 2023, 38 (7): 691- 703.
16	Vonk NA, Channick R, Cottreel E, et al. The REPAIR study: effects of macitentan on RV structure and function in pulmonary arterial hypertension[J]. JACC Cardiovasc Imaging, 2022, 15 (2): 240- 253.
17	Gutiérrez A, Gómez DA, Contreras C, et al. Calcium handling coupled to the endothelin ETA and ETB receptor-mediated vasoconstriction in resistance arteries: differential regulation by PI3K, PKC and RhoK[J]. Eur J Pharmacol, 2023, 956, 175948. doi: 10.1016/j.ejphar.2023.175948
18	Zhou B, Gitschier J. hCTR1: a human gene for copper uptake identified by complementation in yeast[J]. Proc Natl Acad Sci U S A, 1997, 94 (14): 7481- 7486.
19	Wen MH, Chen HH, Yan GJ, et al. Elevated intracellular copper induces CTR1 monomerization and prevents copper uptake[J]. Nat Commun, 2025, 16 (1): 11500. doi: 10.1038/s41467-025-66283-w
20	朱明强, 谢星, 廖启成, 等. 铜死亡的发生机制及在肝脏疾病中的作用[J]. 临床肝胆病杂志, 2024, 40 (11): 2332- 2337.
21	Sheftel AD, Stehling O, Pierik AJ, et al. Humans possess two mitochondrial ferredoxins, FDX1 and FDX2, with distinct roles in steroidogenesis, heme, and Fe/S cluster biosynthesis[J]. Proc Natl Acad Sci U S A, 2010, 107 (26): 11775- 11780. doi: 10.1073/pnas.1004250107
22	Dreishpoon MB, Bick NR, Petrova B, et al. FDX1 regulates cellular protein lipoylation through direct binding to LIAS[J]. J Biol Chem, 2023, 299 (9): 105046. doi: 10.1016/j.jbc.2023.105046
23	任俏慧, 朱欣悦, 吕伟, 等. 伊利司莫-铜离子诱导肝癌细胞系PLC/PRF/5和Huh-7发生铜死亡[J]. 安徽医科大学学报, 2025, 60 (8): 1470- 1477. doi: 10.19405/j.cnki.issn1000-1492.2025.08.015
24	Lairion F, Saporito-Magrina C, Musacco-Sebio R, et al. Nitric oxide, chronic iron and copper overloads and regulation of redox homeostasis in rat liver[J]. J Biol Inorg Chem, 2022, 27 (1): 23- 36. doi: 10.1007/s00775-021-01908-1
25	Klecker T, Westermann B. Pathways shaping the mitochondrial inner membrane[J]. Open Biol, 2021, 11 (12): 210238. doi: 10.1098/rsob.210238
26	Thurakkal B, Hari K, Marwaha R, et al. Collective heterogeneity of mitochondrial potential in contact inhibition of proliferation[J]. Biophys J, 2023, 122 (19): 3909- 3923. doi: 10.1016/j.bpj.2023.08.014
27	汪鸣, 赵信科, 刘生发, 等. 当归红芪超滤物含药血清对放射性H9C2心肌细胞铜死亡的干预效应[J]. 四川中医, 2025, 43 (3): 138- 143.
28	Zhuang R, Zhou Q, Cheng B, et al. PDHA1-acetylation signaling suppresses cuproptosis to attenuate anti-androgen effect in prostate cancer[J]. Cell Death Dis, 2026, 17 (2): 245. doi: 10.1038/s41419-026-08462-1

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硫化氢对低氧性肺动脉高压大鼠FDX1-铜死亡途径的干预

Intervention of hydrogen sulfide on FDX1-cuproptosis pathway in rats with hypoxemic pulmonary hypertension

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