The right choice of antihypertensives protects primary human hepatocytes from ethanol- and recombinant human TGF-β1-induced cellular damage.
BACKGROUND
Patients with alcoholic liver disease (ALD) often suffer from high blood pressure and rely on antihypertensive treatment. Certain antihypertensives may influence progression of chronic liver disease. Therefore, the aim of this study is to investigate the impact of the commonly used antihypertensives amlodipine, captopril, furosemide, metoprolol, propranolol, and spironolactone on alcohol-induced damage toward human hepatocytes (hHeps).


METHODS
hHeps were isolated by collagenase perfusion. Reactive oxygen species (ROS) were measured by fluorescence-based assays. Cellular damage was determined by lactate-dehydrogenase (LDH)-leakage. Expression analysis was performed by reverse-transcription polymerase chain reaction and Western blot. Transforming growth factor (TGF)-β signaling was investigated by a Smad3/4-responsive luciferase-reporter assay.


RESULTS
Ethanol and TGF-β1 rapidly increased ROS in hHeps, causing a release of 40%-60% of total LDH after 72 hours. All antihypertensives dose dependently reduced ethanol-mediated oxidative stress and cellular damage. Similar results were observed for TGF-β1-dependent damage, except for furosemide, which had no effect. As a common mechanism, all antihypertensives increased heme-oxygenase-1 (HO-1) expression, and inhibition of HO-1 activity reversed the protective effect of the drugs. Interestingly, Smad3/4 signaling was reduced by all compounds except furosemide, which even enhanced this profibrotic signaling. This effect was mediated by expressional changes of Smad3 and/or Smad4.


CONCLUSIONS
Our results suggest that antihypertensives may both positively and negatively influence chronic liver disease progression. Therefore, we propose that in future patients with ALD and high blood pressure, they could benefit from an adjusted antihypertensive therapy with additional antifibrotic effects.
Eberhard Karls Universität Tübingen, BG Trauma Center, Tübingen, Germany.
Sabrina
S
Ehnert
Mol Hepatology - Alcohol Associated Diseases, Department of Medicine II, Medical Faculty, Mannheim, Germany.
Teresa
T
Lukoschek
Mol Hepatology - Alcohol Associated Diseases, Department of Medicine II, Medical Faculty, Mannheim, Germany.
Anastasia
A
Bachmann
Eberhard Karls Universität Tübingen, BG Trauma Center, Tübingen, Germany.
Juan J
JJ
Martínez Sánchez
Department of General, Visceral, and Transplantation Surgery, Charité University Medicine, Berlin, Germany.
Georg
G
Damm
Clinic for General, Visceral, Endocrine Surgery and Coloproctology, Clinic Neuperlach, Städtisches Klinikum München GmbH, Munich, Germany.
Natascha C
NC
Nussler
Department of Diabetology, Klinikum Traunstein, Kliniken Südostbayern AG, Traunstein, Germany.
Stefan
S
Pscherer
Eberhard Karls Universität Tübingen, BG Trauma Center, Tübingen, Germany.
Ulrich
U
Stöckle
Mol Hepatology - Alcohol Associated Diseases, Department of Medicine II, Medical Faculty, Mannheim, Germany.
Steven
S
Dooley
Department of Medicine, Salem Medical Center, Ruprecht-Karls-Universität, Heidelberg, Germany.
Sebastian
S
Mueller
Eberhard Karls Universität Tübingen, BG Trauma Center, Tübingen, Germany.
Andreas K
AK
Nussler
TGF-β1; alcoholic liver disease; antihypertensives; ethanol; primary human hepatocytes