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Bioluminescence Imaging of NADPH Oxidase Activity in Different Animal Models
Published on: October 22, 2012
Zymosan-induced luminol-dependent chemiluminescence response of circulating and extravasated leukocytes in
1Research Center, Riyadh Al Kharj Hospital Program, Saudi Arabia. khan_haseeb@yahoo.com
Abstract:
This study examines a concurrent profiling of circulating and extravasated polymorphonuclear leukocytes (PMNs) in a rat model of experimental sepsis. Fecal peritonitis was induced in Wistar male rats by intraperitoneal instillation of a fecal suspension in saline (1:1 w/v). Blood and peritoneal fluid were collected 8 h following fecal inoculation for the evaluation of inflammatory response of PMNs using zymosan-induced luminol-dependent chemiluminescence. Fifty microliters of pre-diluted blood or peritoneal fluid samples were mixed with 150 microl of reaction mixture (4 x 10(-4) M luminol+50 microg opsonized zymosan+0.1% gelatin in Hank's balanced salt solution) and the chemiluminescence signal was measured in a luminometer at 37 degrees C. Fecal peritonitis caused a significant leukocytopenia (3540+/-297 mm(-3) versus control value of 7525+/-711 mm(-3), p < 0.001) accompanied by massive infiltration of PMNs in the peritoneal cavity (34700+/-4006 versus 7325+/-425 mm(-3), p < 0.001). The phagocytic activity of circulating blood PMNs was down-regulated whereas a significant up-regulation was observed in the activity of PMNs from peritoneal fluid. In conclusion, this study clearly demonstrates sepsis-induced alterations in both blood and peritoneal fluid PMNs and their quantitative assessment may be helpful in disease evaluation and designing effective therapies.
Insights
Sepsis significantly alters polymorphonuclear leukocytes (PMNs) in rats, reducing circulating PMN activity while increasing peritoneal PMN activity. This profiling aids in evaluating sepsis and developing therapies.
Area of Science:
- Immunology
- Pathophysiology
- Animal Models
Background:
- Sepsis involves complex inflammatory responses.
- Polymorphonuclear leukocytes (PMNs) are crucial in innate immunity and sepsis pathogenesis.
- Understanding PMN behavior in different body compartments during sepsis is vital.
Purpose of the Study:
- To concurrently profile circulating and extravasated polymorphonuclear leukocytes (PMNs) in a rat model of experimental sepsis.
- To evaluate the inflammatory response and phagocytic activity of PMNs in blood and peritoneal fluid during sepsis.
Main Methods:
- Experimental sepsis induced via fecal peritonitis in Wistar male rats.
- Collection of blood and peritoneal fluid 8 hours post-inoculation.
- Measurement of PMN inflammatory response using zymosan-induced luminol-dependent chemiluminescence.
Main Results:
- Sepsis induced significant leukocytopenia in circulating blood.
- Massive infiltration of PMNs into the peritoneal cavity was observed.
- Circulating PMN phagocytic activity was down-regulated, while peritoneal PMN activity was significantly up-regulated.
Conclusions:
- Sepsis causes distinct alterations in both circulating and extravasated PMNs.
- Quantitative assessment of PMNs in different compartments can aid in sepsis evaluation.
- Findings may inform the development of effective sepsis therapies.
