Digoxin improves steatohepatitis with differential involvement of liver cell subsets in mice through inhibition of PKM2 transactivation

Peng Zhao, Sheng Na Han, Suyavaran Arumugam, Muhammad Nadeem Yousaf, Yanqin Qin, Joy X. Jiang, Natalie Julia Torok, Yonglin Chen, Mohd Salah Mankash, Junbao Liu, Jiansheng Li, Yasuko Iwakiri, Xinshou Ouyang

Research output: Contribution to journalArticle

Abstract

The cardiac glycoside digoxin was identified as a potent suppressor of pyruvate kinase isoform 2-hypoxia-inducible factor-α (PKM2-HIF-1α) pathway activation in liver injury mouse models via intraperitoneal injection. We have assessed the therapeutic effects of digoxin to reduce nonalcoholic steatohepatitis (NASH) by the clinically relevant oral route in mice and analyzed the cellular basis for this effect with differential involvement of liver cell subsets. C57BL/6J male mice were placed on a high-fat diet (HFD) for 10 wk and started concurrently with the gavage of digoxin (2.5, 0.5, 0.125 mg/kg twice a week) for 5 wk. Digoxin significantly reduced HFD-induced hepatic damage, steatosis, and liver inflammation across a wide dosage range. The lowest dose of digoxin (0.125 mg/kg) showed significant protective effects against liver injury and sterile inflammation. Consistently, digoxin attenuated HIF-1α sustained NLRP3 inflammasome activation in macrophages. We have reported for the first time that PKM2 is upregulated in hepatocytes with hepatic steatosis, and digoxin directly improved hepatocyte mitochondrial dysfunction and steatosis. Mechanistically, digoxin directly bound to PKM2 and inhibited PKM2 targeting HIF-1α transactivation without affecting PKM2 enzyme activation. Thus, oral digoxin showed potential to therapeutically inhibit liver injury in NASH through the regulation of PKM2-HIF-1α pathway activation with involvement of multiple cell types. Because of the large clinical experience with oral digoxin, this may have significant clinical applicability in human NASH.NEW & NOTEWORTHY This study is the first to assess the therapeutic efficacy of oral digoxin on nonalcoholic steatohepatitis (NASH) in a high-fat diet (HFD) mouse model and to determine the divergent of cell type-specific effects. Oral digoxin reduced liver damage, steatosis, and inflammation in HFD mice. Digoxin attenuated hypoxia-inducible factor (HIF)-1α axis-sustained inflammasome activity in macrophages and hepatic oxidative stress response in hepatocytes via the regulation of PKM2-HIF-1α axis pathway activation. Oral digoxin may have significant clinical applicability in human NASH.

Original languageEnglish (US)
Pages (from-to)G387-G397
JournalAmerican journal of physiology. Gastrointestinal and liver physiology
Volume317
Issue number4
DOIs
StatePublished - Oct 1 2019

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Digoxin
Fatty Liver
Transcriptional Activation
Liver
Hypoxia-Inducible Factor 1
High Fat Diet
Inflammasomes
Hepatocytes
Inflammation
Wounds and Injuries
Cardiac Glycosides
Enzyme Activation
Pyruvate Kinase
Macrophage Activation
Therapeutic Uses
Intraperitoneal Injections
Protein Isoforms
Oxidative Stress

Keywords

  • liver
  • oral digoxin
  • pyruvate kinase M2-HIF-1α axis
  • steatohepatitis
  • sterile inflammation

ASJC Scopus subject areas

  • Physiology
  • Hepatology
  • Gastroenterology
  • Physiology (medical)

Cite this

Digoxin improves steatohepatitis with differential involvement of liver cell subsets in mice through inhibition of PKM2 transactivation. / Zhao, Peng; Han, Sheng Na; Arumugam, Suyavaran; Yousaf, Muhammad Nadeem; Qin, Yanqin; Jiang, Joy X.; Torok, Natalie Julia; Chen, Yonglin; Mankash, Mohd Salah; Liu, Junbao; Li, Jiansheng; Iwakiri, Yasuko; Ouyang, Xinshou.

In: American journal of physiology. Gastrointestinal and liver physiology, Vol. 317, No. 4, 01.10.2019, p. G387-G397.

Research output: Contribution to journalArticle

Zhao, Peng ; Han, Sheng Na ; Arumugam, Suyavaran ; Yousaf, Muhammad Nadeem ; Qin, Yanqin ; Jiang, Joy X. ; Torok, Natalie Julia ; Chen, Yonglin ; Mankash, Mohd Salah ; Liu, Junbao ; Li, Jiansheng ; Iwakiri, Yasuko ; Ouyang, Xinshou. / Digoxin improves steatohepatitis with differential involvement of liver cell subsets in mice through inhibition of PKM2 transactivation. In: American journal of physiology. Gastrointestinal and liver physiology. 2019 ; Vol. 317, No. 4. pp. G387-G397.
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AU - Han, Sheng Na

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AU - Qin, Yanqin

AU - Jiang, Joy X.

AU - Torok, Natalie Julia

AU - Chen, Yonglin

AU - Mankash, Mohd Salah

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AU - Iwakiri, Yasuko

AU - Ouyang, Xinshou

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N2 - The cardiac glycoside digoxin was identified as a potent suppressor of pyruvate kinase isoform 2-hypoxia-inducible factor-α (PKM2-HIF-1α) pathway activation in liver injury mouse models via intraperitoneal injection. We have assessed the therapeutic effects of digoxin to reduce nonalcoholic steatohepatitis (NASH) by the clinically relevant oral route in mice and analyzed the cellular basis for this effect with differential involvement of liver cell subsets. C57BL/6J male mice were placed on a high-fat diet (HFD) for 10 wk and started concurrently with the gavage of digoxin (2.5, 0.5, 0.125 mg/kg twice a week) for 5 wk. Digoxin significantly reduced HFD-induced hepatic damage, steatosis, and liver inflammation across a wide dosage range. The lowest dose of digoxin (0.125 mg/kg) showed significant protective effects against liver injury and sterile inflammation. Consistently, digoxin attenuated HIF-1α sustained NLRP3 inflammasome activation in macrophages. We have reported for the first time that PKM2 is upregulated in hepatocytes with hepatic steatosis, and digoxin directly improved hepatocyte mitochondrial dysfunction and steatosis. Mechanistically, digoxin directly bound to PKM2 and inhibited PKM2 targeting HIF-1α transactivation without affecting PKM2 enzyme activation. Thus, oral digoxin showed potential to therapeutically inhibit liver injury in NASH through the regulation of PKM2-HIF-1α pathway activation with involvement of multiple cell types. Because of the large clinical experience with oral digoxin, this may have significant clinical applicability in human NASH.NEW & NOTEWORTHY This study is the first to assess the therapeutic efficacy of oral digoxin on nonalcoholic steatohepatitis (NASH) in a high-fat diet (HFD) mouse model and to determine the divergent of cell type-specific effects. Oral digoxin reduced liver damage, steatosis, and inflammation in HFD mice. Digoxin attenuated hypoxia-inducible factor (HIF)-1α axis-sustained inflammasome activity in macrophages and hepatic oxidative stress response in hepatocytes via the regulation of PKM2-HIF-1α axis pathway activation. Oral digoxin may have significant clinical applicability in human NASH.

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