Nicotine-Induced Apoptosis in Human Renal Proximal Tubular Epithelial Cells.
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.Human proximal tubular epithelial (HK-2) cells were treated wi...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2016-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4814027?pdf=render |
| _version_ | 1828159418717437952 |
|---|---|
| author | Chang Seong Kim Joon Seok Choi Soo Yeon Joo Eun Hui Bae Seong Kwon Ma JongUn Lee Soo Wan Kim |
| author_facet | Chang Seong Kim Joon Seok Choi Soo Yeon Joo Eun Hui Bae Seong Kwon Ma JongUn Lee Soo Wan Kim |
| author_sort | Chang Seong Kim |
| collection | DOAJ |
| description | 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.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.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.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. |
| first_indexed | 2024-04-12T00:00:51Z |
| format | Article |
| id | doaj.art-8ed428de6e194cf3833edb5467c04d5f |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-12T00:00:51Z |
| publishDate | 2016-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-8ed428de6e194cf3833edb5467c04d5f2022-12-22T03:56:14ZengPublic Library of Science (PLoS)PLoS ONE1932-62032016-01-01113e015259110.1371/journal.pone.0152591Nicotine-Induced Apoptosis in Human Renal Proximal Tubular Epithelial Cells.Chang Seong KimJoon Seok ChoiSoo Yeon JooEun Hui BaeSeong Kwon MaJongUn LeeSoo Wan KimNicotine 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.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.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.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.http://europepmc.org/articles/PMC4814027?pdf=render |
| spellingShingle | Chang Seong Kim Joon Seok Choi Soo Yeon Joo Eun Hui Bae Seong Kwon Ma JongUn Lee Soo Wan Kim Nicotine-Induced Apoptosis in Human Renal Proximal Tubular Epithelial Cells. PLoS ONE |
| title | Nicotine-Induced Apoptosis in Human Renal Proximal Tubular Epithelial Cells. |
| title_full | Nicotine-Induced Apoptosis in Human Renal Proximal Tubular Epithelial Cells. |
| title_fullStr | Nicotine-Induced Apoptosis in Human Renal Proximal Tubular Epithelial Cells. |
| title_full_unstemmed | Nicotine-Induced Apoptosis in Human Renal Proximal Tubular Epithelial Cells. |
| title_short | Nicotine-Induced Apoptosis in Human Renal Proximal Tubular Epithelial Cells. |
| title_sort | nicotine induced apoptosis in human renal proximal tubular epithelial cells |
| url | http://europepmc.org/articles/PMC4814027?pdf=render |
| work_keys_str_mv | AT changseongkim nicotineinducedapoptosisinhumanrenalproximaltubularepithelialcells AT joonseokchoi nicotineinducedapoptosisinhumanrenalproximaltubularepithelialcells AT sooyeonjoo nicotineinducedapoptosisinhumanrenalproximaltubularepithelialcells AT eunhuibae nicotineinducedapoptosisinhumanrenalproximaltubularepithelialcells AT seongkwonma nicotineinducedapoptosisinhumanrenalproximaltubularepithelialcells AT jongunlee nicotineinducedapoptosisinhumanrenalproximaltubularepithelialcells AT soowankim nicotineinducedapoptosisinhumanrenalproximaltubularepithelialcells |