Alteration of some cellular function in amikacin resistant Pseudomonas aeruginosa transfected macrophages: a time
Subhankari Prasad Chakraborty1, Santanu Karmahapatra, Sabyasachi Das
1Immunology and Microbiology Laboratory, Department of Human Physiology with Community Health, Vidyasagar University, Midnapore-721 102, West Bengal, India.
Objective:
To evaluate the free radical generation and antioxidant enzymes status in murine peritoneal macrophage during in vitro amikacin resistant Pseudomonas aeruginosa (ARPA) treatment with different time interval.
Methods:
Peritoneal macrophages were treated with 1×10(8) CFU/mL ARPA cell suspension in vitro for different time interval (1, 2, 3, 6, 12, and 24 h) and super oxide anion generation, NO generation, reduced glutathione level and antioxidant enzymes status were analyzed.
Results:
Super oxide anion generation and NO generation got peak at 12 h, indicating maximal free radical generation through activation of NADPH oxidase in murine peritoneal macrophages during ARPA transfection. Reduced glutathione level and antioxidant enzymes status were decreased significantly (P<0.05) with increasing time of ARPA transfection. All the changes in peritoneal macrophages after 12 h in vitro ARPA transfection had significant difference (P<0.05).
Conclusions:
From this study, it may be summarized that in vitro ARPA infection not only generates excess free radical but also affects the antioxidant system and glutathione cycle in murine peritoneal macrophage.
Insights
Amikacin-resistant Pseudomonas aeruginosa (ARPA) infection in murine macrophages generates excessive free radicals and depletes antioxidant defenses, peaking at 12 hours. This highlights ARPA
Area of Science:
- Immunology
- Microbiology
- Biochemistry
Background:
- Pseudomonas aeruginosa is an opportunistic pathogen known for its resistance to antibiotics.
- Macrophages play a crucial role in the innate immune response against bacterial infections.
- Oxidative stress is implicated in the pathogenesis of various infections.
Purpose of the Study:
- To investigate the impact of amikacin-resistant Pseudomonas aeruginosa (ARPA) on free radical generation in murine peritoneal macrophages.
- To assess the status of antioxidant enzymes and glutathione levels during ARPA treatment in vitro.
- To determine the time-dependent effects of ARPA on macrophage oxidative stress.
Main Methods:
- Murine peritoneal macrophages were exposed to ARPA (1×10^8 CFU/mL) in vitro for varying durations (1-24 hours).
- Analysis included superoxide anion generation, nitric oxide (NO) generation, and reduced glutathione (GSH) levels.
- Antioxidant enzyme activities were also measured to evaluate cellular defense mechanisms.
Main Results:
- Superoxide anion and NO generation peaked at 12 hours post-ARPA exposure, indicating significant free radical production.
- Reduced glutathione levels and antioxidant enzyme activities significantly decreased over time with ARPA treatment (P<0.05).
- Macrophages exhibited significant alterations after 12 hours of in vitro ARPA transfection (P<0.05).
Conclusions:
- In vitro ARPA infection induces substantial free radical generation in murine peritoneal macrophages.
- ARPA significantly impairs the macrophage antioxidant system and glutathione cycle.
- These findings underscore the detrimental effects of ARPA on cellular oxidative balance.
