Related Experiment Video
Updated: Jun 20, 2026
Using Nicotine in a Silica-Exposed Mouse Model to Promote Lung Epithelial-Mesenchymal Transition
Published on: March 3, 2023
In vitro nicotine-induced oxidative stress in mice peritoneal macrophages: a dose-dependent approach
Santanu Kar Mahapatra1, Subhasis Das, Surajit Bhattacharjee
1Department of Human Physiology with Community Health, Immunology and Microbiology Laboratory, Vidyasagar University, West Bengal, India.
Abstract:
The immune cells use reactive oxygen species (ROS) for carrying out their normal functions while an excess amount of ROS can attack cellular components that lead to cell damage. In the present study, peritoneal macrophages (6 x 10(6) cells, >95% viable) isolated from male Swiss mice were treated with nicotine (1 mM, 5 mM, 10 mM, 25 mM, and 50 mM) in vitro for 12 h and the superoxide anion generation, lipid peroxidation, protein oxidation and antioxidant enzymes status were monitored. Maximum superoxide radical generation was found at the dose of 10 mM nicotine. The lipid peroxidation and protein oxidation were increased significantly (p < 0.05) along with the increasing dose of nicotine. The reduced glutathione level, catalase, superoxide dismutase, glutathione peroxidase, and glutathione reductase activities were decreased significantly (p < 0.05), and oxidized glutathione level was increased significantly (p < 0.05) with the increasing dose of the nicotine. From these experiments, it was also observed that all the changes in peritoneal macrophages with 10 mM, 25 mM, and 50 mM nicotine had no significant difference. To observe the effect of nicotine in vivo, this study examined the liver and spleen antioxidant status after nicotine administration (1 mg/kg BW) intraperitoneally in mice and found the diminished SOD activity and GSH level. It may be concluded that nicotine is able to enhance the production of ROS that produced oxidative stress in murine peritoneal macrophages. It also suggested that, 10 mM in vitro nicotine treatment for 12 h is the effective dose.
Insights
Nicotine exposure increases reactive oxygen species (ROS) and oxidative stress in mouse immune cells. A 10 mM nicotine dose in vitro for 12 hours proved most effective in inducing these damaging effects.
Area of Science:
- Immunology
- Biochemistry
- Toxicology
Background:
- Immune cells utilize reactive oxygen species (ROS) for essential functions.
- Excessive ROS can cause cellular damage, leading to oxidative stress.
- Nicotine's impact on immune cell oxidative stress requires investigation.
Purpose of the Study:
- To investigate the effects of nicotine on oxidative stress markers in murine peritoneal macrophages.
- To determine the effective dose of nicotine for inducing oxidative stress in vitro.
- To assess the in vivo impact of nicotine on antioxidant status in mice.
Main Methods:
- Murine peritoneal macrophages were treated with varying nicotine concentrations (1-50 mM) in vitro for 12 hours.
- Assessed superoxide anion generation, lipid peroxidation, and protein oxidation.
- Monitored antioxidant enzyme status (glutathione, catalase, SOD, GPx, GR) and in vivo liver/spleen antioxidant levels.
Main Results:
- Nicotine significantly increased superoxide radical generation, lipid peroxidation, and protein oxidation in a dose-dependent manner, with maximum effect at 10 mM.
- Reduced glutathione levels and antioxidant enzyme activities (catalase, SOD, GPx, GR) decreased significantly with increasing nicotine doses.
- In vivo nicotine administration diminished superoxide dismutase (SOD) activity and reduced glutathione (GSH) levels in mouse liver and spleen.
Conclusions:
- Nicotine enhances ROS production, inducing significant oxidative stress in murine peritoneal macrophages.
- A 10 mM in vitro nicotine treatment for 12 hours is identified as an effective dose for inducing oxidative stress.
- Nicotine exposure negatively impacts antioxidant status both in vitro and in vivo.
