Characterization of Acetaminophen Toxicity in Human Kidney HK-2 Cells

Abstract

Acetaminophen (APAP) overdose causes liver injury, but in some cases it is associated also with renal impairment. While several studies exist in relation to acetaminophen nephrotoxicity, no reports have been published describing intracellular changes related to APAP nephrotoxicity in vitro. Because proximal tubular cells are considered to constitute a secondary site of drug-induced injury after hepatocytes, our study's aim was to estimate the toxicity in the human HK-2 cell line. We used a range of APAP concentrations (1-10 mM) to examine toxicity in the cells (1-48 h). We evaluated cell viability using the WST-1 and LDH tests. Cells impairment was also determined by monitoring ROS production, glutathione levels. We proved that HK-2 cells are able to metabolize acetaminophen. We observed moderate impairment of cells already after 1 h of treatment based on a finding of increased ROS production and decreased cell viability. After 24 h, the results showed significant cellular impairment at all tested concentrations except for 1 mM APAP, but no glutathione depletion was found. We conclude that HK-2 cells are susceptible to acetaminophen toxicity but, unlike hepatocytes, it might be not linked to glutathione depletion.

In VitrotoIn VivoConcordance of Toxicity Using the Human Proximal Tubule Cell Line HK-2

“…Although the human proximal tubule cell line, HK-2, has been used in a plethora of studies to help establish the nephrotoxicity potential of select toxins, [11][12][13][14][15][16] there is a lack of comprehensive research focused on the use of the HK-2 cell line as a tool to effectively predict and verify in vivo toxicity on proximal tubule injury. Understanding the concordance between data generated from in vitro cell models, in vivo animal models, and human clinical studies lends credence to in vitro studies in settings where risk assessments of potential toxins are being pursued.…”

Isolated paracetamol-induced acute kidney injury: a systematic review

“…Although paracetamol requires metabolic activation to cause nephrotoxicity, 15 the specific pathways involved are unclear with data consisting largely of animal studies. Potential roles of cytochrome P450 metabolites and glutathione depletion have been identified, 16 suggesting that conditions predisposing to hepatotoxicity may similarly enhance nephrotoxicity. 1 However, evidence of cytochrome P450-independent mechanisms also exists.…”

Transient Increase in Cellular Dehydrogenase Activity After Cadmium Treatment Precedes Enhanced Production of Reactive Oxygen Species in Human Proximal Tubular Kidney Cells

“…Human kidney (HK-2) cells, a proximal tubular epithelial cell line derived from normal adult human kidney cells immortalized by transduction with human papillomavirus (HPV 16) DNA fragment (Ryan et al 1994), were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). The cells were cultured in keeping with our previous studies (Hauschke et al 2017, Vrbová et al 2016. All the experiments were conducted using the HK-2 cells (passages 4-11).…”