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Published on: January 7, 2013
Nicotine-Induced Apoptosis in Human Renal Proximal Tubular Epithelial Cells
Chang Seong Kim1, Joon Seok Choi1, Soo Yeon Joo1,2
1Department of Internal Medicine, Chonnam National University Medical School, Gwangju, Korea.
Background:
Nicotine is, to a large extent, responsible for smoking-mediated renal dysfunction. This study investigated nicotine's effects on renal tubular epithelial cell apoptosis in vitro and it explored the mechanisms underlying its effects.
Methods:
Human proximal tubular epithelial (HK-2) cells were treated with nicotine. Cell viability was examined by using the WST-1 assay. Intracellular levels of reactive oxygen species (ROS) and the expression of mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) proteins were determined. The messenger ribonucleic acid and the protein expression associated with the nicotine acetylcholine receptors (nAChRs) in HK-2 cells was examined, and apoptosis was detected using flow cytometry, cell cycle analysis, and immunoblot analysis.
Results:
The HK-2 cells were endowed with nAChRs. Nicotine treatment reduced cell viability dose dependently, increased ROS levels, and increased extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 MAPK expression. Nicotine increased NF-κB activation, which was attenuated by N-acetyl-L-cysteine, and ERK and JNK inhibitors, but was not affected by a p38 MAPK inhibitor. Nicotine increased the Bax/Bcl-2 ratio, which was attenuated by N-acetyl-L-cysteine, the NF-κB inhibitor, Bay 11-7082, and hexamethonium, a non-specific nAChR blocker. Flow cytometry revealed nicotine-induced G2/M phase arrest. While nicotine treatment increased the expression of phosphorylated cdc2 and histone H3, a marker of G2/M phase arrest, hexamethonium and Bay 11-7082 pretreatment reduced their expression.
Conclusions:
Nicotine caused apoptosis in HK-2 cells by inducing ROS generation that activated the NF-κB signaling pathway via the MAPK pathway and it arrested the cell cycle at the G2/M phase. Nicotine-induced apoptosis in HK-2 cells involves the nAChRs.
Insights
Nicotine induces kidney cell apoptosis by increasing reactive oxygen species and activating signaling pathways. This leads to cell cycle arrest, highlighting nicotine
Area of Science:
- Nephrology
- Cell Biology
- Toxicology
Background:
- Smoking is a major cause of kidney dysfunction.
- Nicotine is the primary component of tobacco smoke responsible for these effects.
- This study examines nicotine's impact on kidney cells.
Purpose of the Study:
- To investigate the effects of nicotine on renal tubular epithelial cell apoptosis in vitro.
- To elucidate the underlying mechanisms of nicotine-induced renal cell apoptosis.
Main Methods:
- Human proximal tubular epithelial (HK-2) cells were treated with nicotine.
- Assessed cell viability, reactive oxygen species (ROS) levels, and MAPK/NF-κB signaling.
- Examined nicotine acetylcholine receptors (nAChRs) and apoptosis markers.
Main Results:
- Nicotine reduced cell viability and increased ROS generation.
- Nicotine activated MAPK (ERK, JNK, p38) and NF-κB pathways.
- Nicotine induced G2/M cell cycle arrest and apoptosis, involving nAChRs.
Conclusions:
- Nicotine induces apoptosis in kidney cells via ROS generation and subsequent activation of NF-κB and MAPK signaling pathways.
- Nicotine causes G2/M cell cycle arrest in renal tubular epithelial cells.
- Nicotine acetylcholine receptors play a crucial role in nicotine-induced renal cell apoptosis.
