Pharmacological inhibition of ENaC or NCX can attenuate hepatic ischemia-reperfusion injury exacerbated by hypernatremia

Yabin CHEN; Hao LI; Peihao WEN; Jiakai ZHANG; Zhihui WANG; Shengli CAO; Wenzhi GUO

doi:10.1631/jzus.B2300825

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Research Article | Updated：2025-05-28

Pharmacological inhibition of ENaC or NCX can attenuate hepatic ischemia-reperfusion injury exacerbated by hypernatremia
Pharmacological inhibition of ENaC or NCX can attenuate hepatic ischemia-reperfusion injury exacerbated by hypernatremia
药物抑制ENaC或NCX可减轻高钠血症加重的肝脏缺血再灌注损伤
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology) Vol. 26, Issue 5, Pages: 461-476(2025)
Affiliations：

1.Department of Hepatobiliary and Pancreatic Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China
2.Henan Organ Transplantation Centre, Zhengzhou 450052, China
Funds：
DOI：10.1631/jzus.B2300825
CLC：
CSTR：
Received：14 November 2023，

Revised：22 April 2024，

Published：28 May 2025
Accepted：

陈亚斌,李豪,温培豪等.药物抑制ENaC或NCX可减轻高钠血症加重的肝脏缺血再灌注损伤[J].浙江大学学报（英文版）（B辑：生物医学和生物技术）,2025,26(05):461-476.

Yabin CHEN, Hao LI, Peihao WEN, et al. Pharmacological inhibition of ENaC or NCX can attenuate hepatic ischemia-reperfusion injury exacerbated by hypernatremia[J]. Journal of zhejiang university-science b (biomedicine & biotechnology), 2025, 26(5): 461-476.
陈亚斌,李豪,温培豪等.药物抑制ENaC或NCX可减轻高钠血症加重的肝脏缺血再灌注损伤[J].浙江大学学报（英文版）（B辑：生物医学和生物技术）,2025,26(05):461-476. DOI： 10.1631/jzus.B2300825.

Yabin CHEN, Hao LI, Peihao WEN, et al. Pharmacological inhibition of ENaC or NCX can attenuate hepatic ischemia-reperfusion injury exacerbated by hypernatremia[J]. Journal of zhejiang university-science b (biomedicine & biotechnology), 2025, 26(5): 461-476. DOI： 10.1631/jzus.B2300825.

摘要

血清钠离子浓度

155 mmol/L的供体是肝移植（LT）的扩展标准供体。供体血钠升高将导致肝移植术后早期肝功能不良的发生率增加，但其确切机制尚未见报道。本研究建立了70%肝脏缺血再灌注（I/R）的Lewis大鼠高钠血症模型和高钠（HS）培养基预处理的BRL-3A细胞缺氧复氧（H/R）模型。为了确定损伤程度，本研究进行了生物化学分析、组织学分析以及氧化应激和细胞凋亡指标的检测。本研究应用上皮钠离子通道（ENaC）和Na

/Ca

交换体（NCX）的特异性抑制剂，在体内和体外实验中验证二者在损伤中的作用。HS+I/R组血清丙氨酸氨基转移酶（ALT）、天门冬氨酸氨基转移酶（AST）、乳酸脱氢酶（LDH）水平和肝脏坏死面积均显著升高，活性氧（ROS）生成增多，MPO阳性细胞增多，细胞凋亡加重。与H/R组相比，HS+H/R组BRL-3A细胞凋亡和ROS产生显著增加。应用ENaC的特异性抑制剂阿米洛利（amiloride），在体内和体外均可减轻HS加重的缺血再灌注损伤，具体表现为血清转氨酶、炎性细胞因子、细胞凋亡和氧化应激的降低。NCX的特异性抑制剂SN-6也有类似的作用。综上所述，高钠血症加重肝脏缺血再灌注损伤，但可以通过药物抑制ENaC或NCX来减轻这种损伤作用。

Abstract

Donors with a serum sodium concentration of

155 mmol/L are extended criteria donors for liver transplantation (LT). Elevated serum sodium of donors leads to an increased incidence of hepatic dysfunction in the early postoperative period of LT; however

the exact mechanism has not been reported. We constructed a Lewis rat model of 70% hepatic parenchymal area subjected to ischemia-reperfusion (I/R) with hypernatremia and a BRL-3A cell model of hypoxia-reoxygenation (H/R) with high-sodium (HS) culture medium precondition. To determine the degree of injury

biochemical analysis

histological analysis

and oxidative stress and apoptosis detection were performed. We applied specific inhibitors of the epithelial sodium channel (ENaC) and Na

/Ca

exchanger (NCX) in vivo and in vitro to verify their roles in injury. Serum alanine aminotransferase (ALT)

aspartate aminotransferase (AST)

and lactate dehydrogenase (LDH) levels and the area of hepatic necrosis were significantly e

levated in the HS+I/R group. Increased reactive oxygen species (ROS) production

myeloperoxidase (MPO)‍-positive cells

and aggravated cellular apoptosis were detected in the HS+I/R group. The HS+H/R group of BRL-3A cells showed significantly increased cellular apoptosis and ROS production compared to the H/R group. The application of amiloride (Amil)

a specific inhibitor of ENaC

reduced ischemia-reperfusion injury (IRI) aggravated by HS both in vivo and in vitro

as evidenced by decreased serum transaminases

inflammatory cytokines

apoptosis

and oxidative stress. SN-6

a specific inhibitor of NCX

had a similar effect to Amil. In summary

hypernatremia aggravates hepatic IRI

which can be attenuated by pharmacological inhibition of ENaC or NCX.

关键词

Keywords

references

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