Procalcitonin Impairs Liver Cell Viability and Function In Vitro: A Potential New Mechanism of Liver Dysfunction and Failure during Sepsis?

Martin Sauer,Nana-Maria Wagner,Sandra Doß,Johannes Ehler,Thomas Mencke

doi:10.1155/2017/6130725

Abstract

Purpose. Liver dysfunction and failure are severe complications of sepsis and result in poor outcome and increased mortality. The underlying pathologic mechanisms of hepatocyte dysfunction and necrosis during sepsis are only incompletely understood. Here, we investigated whether procalcitonin, a biomarker of sepsis, modulates liver cell function and viability. Materials and Methods. Employing a previously characterized and patented biosensor system evaluating hepatocyte toxicity in vitro, human hepatocellular carcinoma cells (HepG2/C3A) were exposed to 0.01–50 ng/mL procalcitonin for 2 × 72 h and evaluated for proliferation, necrosis, metabolic activity, cellular integrity, microalbumin synthesis, and detoxification capacity. Acetaminophen served as positive control. For further standardization, procalcitonin effects were confirmed in a cellular toxicology assay panel employing L929 fibroblasts. Data were analyzed using ANOVA/Tukey's test. Results. Already at concentrations as low as 0.25 ng/mL, procalcitonin induced HepG2/C3A necrosis (P < 0.05) and reduced metabolic activity, cellular integrity, synthesis, and detoxification capacity (all P < 0.001). Comparable effects were obtained employing L929 fibroblasts. Conclusion. We provide evidence for procalcitonin to directly impair function and viability of human hepatocytes and exert general cytotoxicity in vitro. Therapeutical targeting of procalcitonin could thus display a novel approach to reduce incidence of liver dysfunction and failure during sepsis and lower morbidity and mortality of septic patients.

Highlights

Sepsis is one of the leading causes of death in critically ill patients admitted to intensive care units [1]
A human hepatocyte cell line HepG2/C3A was obtained from the American Type Culture Collection (ATCC CRL-10741, WCB 25022009) and cultivated in Dulbecco’s modified Eagle’s Medium (Dulbecco’s DMEM, GIBCO Life Technologies, Eggenstein, Germany), supplemented with 10% fetal calf serum (PAA Laboratories, Colbe, Germany), 1% of glutamine solution (PAA), and 1% of antibiotics solution (Penicillin G: 10,000 IE/mL/Streptomycin: 10 mg/mL; PAA)
250,000 hepatocytes were seeded per well of a 24-well plate, incubated in 1 mL medium for 72 h, and exposed to procalcitonin (Sigma-Aldrich, Steinheim am Albuch, Germany) ranging from 0.01 to 50 ng/mL or phosphate buffered saline (PBS) only serving as vehicle control

Summary

Introduction

Sepsis is one of the leading causes of death in critically ill patients admitted to intensive care units [1]. Hepatic dysfunction is a key component of MODS and liver failure occurs in nearly 19% of patients with septic shock. Hepatic dysfunction thereby crucially contributes to poor prognosis and outcome [3]. In the pathophysiology of sepsis, the liver modulates several aspects of disease initiation and progression. It clears bacteria and endotoxin from the splanchnic area and tailors host immune responses by secreting inflammatory mediators [4, 5]. Impairment of liver cell function is one of the most crucial amplifiers of MODS during sepsis, its pathophysiology during sepsis is still incompletely understood [6]

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