Nevirapine induces apoptosis in liver (HepG2) cells

Jeerang Wongtrakul; Atchara Paemanee; Phitchayapak Wintachai; Chutima Thepparit; Sittiruk Roytrakul; Thananya Thongtan; Kanokwan Janphen; Khuanchai Supparatpinyo; Duncan R Smith

doi:10.1016/j.apjtm.2016.04.015

Nevirapine induces apoptosis in liver (HepG2) cells

Asian Pac J Trop Med. 2016 Jun;9(6):547-53. doi: 10.1016/j.apjtm.2016.04.015. Epub 2016 Apr 16.

Authors

Jeerang Wongtrakul¹, Atchara Paemanee², Phitchayapak Wintachai², Chutima Thepparit², Sittiruk Roytrakul³, Thananya Thongtan⁴, Kanokwan Janphen¹, Khuanchai Supparatpinyo⁵, Duncan R Smith⁶

Affiliations

¹ Research Institute for Health Sciences, Chiang Mai University, 110 Intavaroros Road, Sriphum, Muang District, Chiang Mai 50200, Thailand.
² Molecular Pathology Laboratory, Institute of Molecular Biosciences, Mahidol University, 25/25 Phuttamonthol Sai 4, Salaya, Nakorn Pathom 73170, Thailand.
³ Proteomics Research Laboratory, Genome Institute, National Center for Genetic Engineering and Biotechnology, 113 Thailand Science Park, Phahonyothin Road, Khlong Nueng, Khlong Luang, Pathumthani 12120, Thailand.
⁴ Department of Biochemistry, Faculty of Medicine, Chulalongkorn University, 1873 Rama 4 Road, Pathumwan, Bangkok 10330, Thailand.
⁵ Department of Medicine, Faculty of Medicine, Chiang Mai University, 239 Huay Kaew Road, Muang District, Chiang Mai 50200, Thailand.
⁶ Molecular Pathology Laboratory, Institute of Molecular Biosciences, Mahidol University, 25/25 Phuttamonthol Sai 4, Salaya, Nakorn Pathom 73170, Thailand. Electronic address: [email protected].

PMID: 27262065
DOI: 10.1016/j.apjtm.2016.04.015

Abstract

Objective: To generate insights into the mechanism of NVP induced hepatotoxicity.

Methods: Liver (HepG2) cells were cultured with various concentrations of NVP. This cell line was chosen because it has low expression of cytochrome P450, allowing evaluation of the effects of NVP rather than specific metabolites. Cytotoxicity was determined using a proliferation assay and cell numbers were monitored using trypan blue exclusion assay for long term culture experiments and apoptosis induction was determined by morphological and biochemical investigation.

Results: HepG2 cells treated with the highest concentration of NVP tested (819 μM) initially showed a rounded morphology and all cells had died by week three of exposure. Nuclear condensation and fragmentation, increased Annexin V/propidium iodide staining and caspase 9 activation all supported the induction of apoptosis in HepG2 cells in response to NVP treatment.

Conclusions: There is a clear induction of apoptosis in response to NVP which suggests that NVP has significant cytotoxicity, over and above any cytotoxicity of metabolites and may contribute directly to patient hepatotoxicity.

Keywords: Apoptosis; Cytotoxicity; Hepatotoxicity; Nevirapine.