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Updated: Sep 27, 2026
Isolation of Murine Peritoneal Macrophages to Carry Out Gene Expression Analysis Upon Toll-like Receptors Stimulation
Published on: April 29, 2015
Peritoneal macrophage activity after laparoscopy or laparotomy
Carmelo Romeo1, Pietro Impellizzeri, Pietro Antonuccio
1Unit of Paediatric Surgery, Department of Medical and Paediatric Surgical Sciences, University of Messina, Messina, Italy.
Background/Purpose:
The beneficial effect of laparoscopy on the immune system seems not to be present at the peritoneal level at which the local immunity appears to be impaired by CO2. The aim of this study was to investigate the peritoneal macrophages (MPhis) activity after laparoscopy and laparotomy.
Methods:
Thirty rats (300 to 350 g) were used and divided into 3 groups. In the laparotomy group, the peritoneum was opened and the abdominal wall exposed to air for 60'. In the laparoscopy group, pneumoperitoneum was created and maintained at 4 to 6 mm Hg for 60'. In the control group, anaesthesia was maintained for 60' without any other manipulations. Twenty-four hours after operation, peritoneal MPhis were harvested and cultured. The authors investigated nitrite/nitrate (NOx) production as well as the message for the inducible nitric oxide (iNOS), with the reverse transcriptase polymerase chain reaction under basal condition and after Lipopolysaccharide (LPS) stimulation.
Results:
Basal MPhi NOx release was significantly higher after laparoscopy than either after laparotomy or control. LPS stimulation produced a strong increase in NOx production in laparotomy MPhis and control MPhis. In contrast, the increase in NOx production was markedly reduced in MPhi harvested after laparoscopy. Similar results were obtained for macrophage mRNA for iNOS; indeed, the increase in iNOS mRNA after LPS stimulation was blunted severely in MPhi from laparoscopy group.
Conclusions:
MPhis after laparoscopy display an higher basal immune performance. After a second insult (LPS), they display a state of tolerance or desensitisation (blunted NOx production and reduced mRNA expression). This observation could have important implications in considering laparoscopy in patients with malignancy or sepsis.
Insights
Laparoscopy impairs peritoneal macrophage immune response, showing reduced nitric oxide production after stimulation. This suggests potential implications for patients with sepsis or malignancy undergoing laparoscopic procedures.
Area of Science:
- Immunology
- Surgical Innovation
- Cellular Biology
Background:
- Laparoscopy's systemic immune benefits may not extend to the peritoneal cavity.
- Carbon dioxide insufflation during laparoscopy may impair local peritoneal immunity.
Purpose of the Study:
- To investigate the activity of peritoneal macrophages (MPhis) following laparoscopic surgery.
- To compare peritoneal MPhis activity after laparoscopy versus traditional laparotomy.
Main Methods:
- Thirty rats were divided into laparoscopy, laparotomy, and control groups.
- Peritoneal macrophages were harvested 24 hours post-procedure.
- Nitrite/nitrate (NOx) production and inducible nitric oxide synthase (iNOS) mRNA expression were assessed before and after lipopolysaccharide (LPS) stimulation.
Main Results:
- Basal NOx release was higher in the laparoscopy group compared to laparotomy and control groups.
- LPS stimulation significantly increased NOx production in laparotomy and control groups.
- LPS stimulation markedly reduced NOx production and iNOS mRNA expression in the laparoscopy group, indicating blunted response.
Conclusions:
- Peritoneal macrophages exhibit enhanced basal immune activity post-laparoscopy.
- Macrophages display a desensitized state after a secondary inflammatory challenge (LPS).
- These findings suggest potential risks for patients with malignancy or sepsis undergoing laparoscopy.
