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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

程缘¹, 田云娜¹, 黄曼¹, 徐俊鹏¹, 曹文傑¹, 贾旭广², 尤利益³, 王万铁^1,*

¹温州医科大学缺血/再灌注损伤研究所，温州 325035；²宜宾学院体育与大健康学院，宜宾 644000；³温州医科大学温州市第三临床学院超声科，温州 325400

摘要

本文旨在探讨硫化氢(hydrogen sulfide, H2S)是否通过抑制有氧糖酵解-细胞焦亡而改善大鼠低氧性肺动脉高压(hypoxic pulmonary hypertension, HPH)。雄性Sprague Dawley (SD)大鼠随机分为正常对照组、正常+NaHS组、低氧组和低氧+NaHS组，每组各6只。正常对照组大鼠置于常氧(21% O2)环境中，每天腹腔注射等体积生理盐水；正常+NaHS组大鼠置于常氧环境中，每天腹腔注射14 μmol/kg NaHS；低氧组大鼠置于缺氧箱中，箱内氧气控制器通过控制N2流速使氧浓度维持在9%~10%，大鼠每天腹腔注射等体积生理盐水；低氧+NaHS组大鼠同样置于缺氧箱中，每天腹腔注射14 μmol/kg NaHS。四周造模结束后，用右心导管插管技术检测各组大鼠的平均肺动脉压(mean pulmonary artery pressure, mPAP)，称重并计算右心室肥厚指数(right ventricular hypertrophy index, RVHI)，用HE染色观察肺组织病理改变程度，用Masson染色观察肺组织纤维化情况，用Western blot检测肺组织己糖激酶2 (hexokinase 2, HK2)、丙酮酸脱氢酶(pyruvate dehydrogenase, PDH)、丙酮酸激酶M2 (pyruvate kinase isozyme type M2, PKM2)、核苷酸结合寡聚化结构域样受体蛋白3 (nucleotide-binding oligomerizationdomain-like receptor protein 3, NLRP3)、GSDMD蛋白N-端片段(GSDMD-N-terminal domain, GSDMD-N)、Caspase-1、白细胞介素-1β (interleukin-1β, IL-1β)和IL-18蛋白表达水平，用ELISA检测肺组织IL-1β和IL-18含量。结果显示，与正常对照组相比，正常+NaHS组各项指标无明显变化，而低氧组mPAP和RVHI显著上升，肺血管壁明显增厚、管腔狭窄，胶原纤维增多，肺组织有氧糖酵解相关蛋白HK2、PKM2蛋白表达显著上调，细胞焦亡相关蛋白NLRP3、GSDMD-N、Caspase-1、IL-1β、IL-18蛋白表达显著上调，IL-1β、IL-18含量增多；低氧组的上述指标的变化均被NaHS显著逆转。上述结果提示，H2S可通过抑制有氧糖酵解-细胞焦亡改善大鼠HPH。

关键词： 硫化氢; 肺动脉高压; 糖酵解; 细胞焦亡; 低氧

Hydrogen sulfide ameliorates hypoxic pulmonary hypertension in rats by inhibiting aerobic glycolysis-pyroptosis

CHENG Yuan¹, TIAN Yun-Na¹, HUANG Man¹, XU Jun-Peng¹, CAO Wen-Jie¹, JIA Xu-Guang², YOU Li-Yi³, WANG Wan-Tie^1,*

¹Institute of Ischemia/Reperfusion Injury, Wenzhou Medical University, Wenzhou 325035, China；²College of Physical Education and Health, Yibin University, Yibin 644000, China；³Department of Ultrasound, Wenzhou Third Clinical Institute Affiliated to Wenzhou Medical University, Wenzhou 325400, China

Abstract

The present study aimed to explore whether hydrogen sulfide (H2S) improved hypoxic pulmonary hypertension (HPH) in rats by inhibiting aerobic glycolysis-pyroptosis. Male Sprague-Dawley (SD) rats were randomly divided into normal group, normal+ NaHS group, hypoxia group, and hypoxia+NaHS group, with 6 rats in each group. The control group rats were placed in a normoxic (21% O2) environment and received daily intraperitoneal injections of an equal volume of normal saline. The normal+NaHS group rats were placed in a normoxic environment and intraperitoneally injected with 14 μmol/kg NaHS daily. The hypoxia group rats were placed in a hypoxia chamber, and the oxygen controller inside the chamber maintained the oxygen concentration at 9% to 10% by controlling the N2 flow rate. An equal volume of normal saline was injected intraperitoneally every day. The hypoxia+NaHS group rats were also placed in an hypoxia chamber and intraperitoneally injected with 14 μmol/kg NaHS daily. After the completion of the four-week modeling, the mean pulmonary artery pressure (mPAP) of each group was measured using right heart catheterization technique, and the right ventricular hypertrophy index (RVHI) was weighed and calculated. HE staining was used to observe pathological changes in lung tissue, Masson staining was used to observe fibrosis of lung tissue, and Western blot was used to detect protein expression levels of hexokinase 2 (HK2), pyruvate dehydrogenase (PDH), pyruvate kinase isozyme type M2 (PKM2), nucleotidebinding oligomerization domain-like receptor protein 3 (NLRP3), GSDMD-N-terminal domain (GSDMD-N), Caspase-1, interleukin- 1β (IL-1β) and IL-18 in lung tissue. ELISA was used to detect contents of IL-1β and IL-18 in lung tissue. The results showed that, compared with the normal control group, there were no significant changes in all indexes in the normal+NaHS group, while the hypoxia group exhibited significantly increased mPAP and RVHI, thickened pulmonary vascular wall, narrowed lumen, increased collagen fibers, up-regulated expression levels of aerobic glycolysis-related proteins (HK2 and PKM2), up-regulated expression levels of pyroptosis-related proteins (NLRP3, GSDMD-N, Caspase-1, IL-1β, and IL-18), and increased contents of IL-1β and IL-18. These changes of the above indexes in the hypoxia group were significantly reversed by NaHS. These results suggest that H2S can improve rat HPH by inhibiting aerobic glycolysis-pyroptosis.

Key words: hydrogen sulfide; pulmonary hypertension; glycolysis; pyroptosis; hypoxia

收稿日期：　　录用日期：

通讯作者：王万铁　　E-mail:

DOI: 10.13294/j.aps.2025.0015

引用本文：

程缘, 田云娜, 黄曼, 徐俊鹏, 曹文傑, 贾旭广, 尤利益, 王万铁. 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.

CHENG Yuan, TIAN Yun-Na, HUANG Man, XU Jun-Peng, CAO Wen-Jie, JIA Xu-Guang, YOU Li-Yi, WANG Wan-Tie. Hydrogen sulfide ameliorates hypoxic pulmonary hypertension in rats by inhibiting aerobic glycolysis-pyroptosis. Acta Physiol Sin 2025; 77 (3): 465-471 (in Chinese with English abstract).