Yonsei Med J.  2016 Jan;57(1):238-246. 10.3349/ymj.2016.57.1.238.

Effect of Pneumoperitoneum on Oxidative Stress and Inflammation via the Arginase Pathway in Rats

• Shin S
• Na S
• Kim OS
• Choi YS
• Kim SH
• Oh YJ

• KMID: 2466376
• DOI: http://doi.org/10.3349/ymj.2016.57.1.238

Abstract

PURPOSE
Oxidative stress during CO2 pneumoperitoneum is reported to be associated with decreased bioactivity of nitric oxide (NO). However, the changes in endothelial nitric oxide synthase (eNOS), inducible nitric oxide synthase (iNOS), and arginase during CO2 pneumoperitoneum have not been elucidated.
MATERIALS AND METHODS
Thirty male Sprague-Dawley rats were randomized into three groups. After anesthesia induction, the abdominal cavities of the rats of groups intra-abdominal pressure (IAP)-10 and IAP-20 were insufflated with CO2 at pressures of 10 mm Hg and 20 mm Hg, respectively, for 2 hours. The rats of group IAP-0 were not insufflated. After deflation, plasma NO was measured, while protein expression levels and activity of eNOS, iNOS, arginase (Arg) I, and Arg II were analyzed with aorta and lung tissue samples.
RESULTS
Plasma nitrite concentration and eNOS expression were significantly suppressed in groups IAP-10 and IAP-20 compared to IAP-0. While expression of iNOS and Arg I were comparable between the three groups, Arg II expression was significantly greater in group IAP-20 than in group IAP-0. Activity of eNOS was significantly lower in groups IAP-10 and IAP-20 than in group IAP-0, while iNOS activity was significantly greater in group IAP-20 than in groups IAP-0 and IAP-10. Arginase activity was significantly greater in group IAP-20 than in groups IAP-0 and IAP-10.
CONCLUSION
The activity of eNOS decreases during CO2 pneumoperitoneum, while iNOS activity is significantly increased, a change that contributes to increased oxidative stress and inflammation. Moreover, arginase expression and activity is increased during CO2 pneumoperitoneum, which seems to act inversely to the NO system.

Keyword

Arginase; nitric oxide synthase; oxidative stress; pneumoperitoneum

MeSH Terms

Animals
Aorta/*physiology
Arginase/*antagonists & inhibitors
Enzyme Inhibitors/administration & dosage/pharmacology
Inflammation/etiology/*prevention & control
Injections, Subcutaneous
Lung Injury/etiology/prevention & control
Male
Nitric Oxide/metabolism
Nitric Oxide Synthase Type II/*metabolism
Nitric Oxide Synthase Type III/*metabolism
Oxidative Stress/*drug effects
Pneumoperitoneum/*complications/drug therapy
Rats
Rats, Sprague-Dawley
Arginase
Enzyme Inhibitors
Nitric Oxide
Nitric Oxide Synthase Type II
Nitric Oxide Synthase Type III

Figure

• 

  Fig. 1 Effect of CO2 pneumoperitoneum at different IAPs on plasma nitrite and tissue MDA concentrations. (A) Plasma nitrite levels were significantly lower in groups IAP-10 and IAP-20 than in group IAP-0. There was no difference between groups IAP-10 and IAP-20. (B) MDA levels were significantly higher in group IAP-20 than in groups IAP-0 and IAP-10. There was no difference between groups IAP-0 and IAP-10. *p<0.01, **p<0.001. MDA, malondialdehyde; IAP, intra-abdominal pressure.

• 

  Fig. 2 Effect of CO2 pneumoperitoneum at different IAPs on eNOS, iNOS, Arg I, and Arg II protein expression. (A) Western blot for eNOS, iNOS, Arg I, Arg II, and β-actin (internal reference) in rat aorta tissue after different intra-abdominal pressures. (B) The expression of eNOS was significantly more suppressed in groups IAP-10 and IAP-20 than in group IAP-0, while there was no difference between groups IAP-10 and IAP-20. (C) There were no significant differences in iNOS expression among the three groups. (D) There were no significant differences in Arg I expression among the three groups. (E) Arg II expression was significantly higher in group IAP-20 than in group IAP-0. *p<0.05, **p<0.01. IAP, intra-abdominal pressure; NOS, nitric oxide synthase; eNOS, endothelial NOS; iNOS, inducible NOS; Arg, arginase.

• 

  Fig. 3 Effect of CO2 pneumoperitoneum at different IAPs on eNOS, iNOS, and arginase activity. (A) The activity of eNOS was significantly lower in groups IAP-10 and IAP-20 than in group IAP-0, with no difference between groups IAP-10 and IAP-20. (B) The activity of iNOS was significantly higher in group IAP-20 than in groups IAP-0 and IAP-10. (C) Arginase activity was significantly higher in group IAP-20 than in groups IAP-0 and IAP-10. No difference was seen between groups IAP-0 and IAP-10. *p<0.05, **p<0.01, ***p<0.001. IAP, intra-abdominal pressure; NOS, nitric oxide synthase; eNOS, endothelial NOS; iNOS, inducible NOS.

• 

  Fig. 4 Effect of CO2 pneumoperitoneum at different IAPs on proinflammatory cytokine concentrations. (A) IL-1β was significantly higher in group IAP-20 than in group IAP-0. (B) IL-6 was significantly higher in group IAP-20 than in groups IAP-0 and IAP-10; however, there was no difference between groups IAP-0 and IAP-10. (C) TNF-α was significantly higher in groups IAP-10 and IAP-20 than in IAP-0, while there was no difference between groups IAP-10 and IAP-20. *p<0.05, **p<0.01. IAP, intra-abdominal pressure; IL, interleukin; TNF, tumor necrosis factor.

• 

  Fig. 5 The effects of different IAPs on histopathological changes in CO2 pneumoperitoneum-induced lung injury in rats. (A) Total scores of lung injury were significantly higher in group IAP-20 than in groups IAP-0 and IAP-10. (B) Normal lung tissue is seen in the IAP-0 group. (C) A slight increase in cellularity (arrows) is seen in the IAP-10 group. (D) Inflammatory cell infiltration (arrows) with protein exudation (asterisks) and alveolus collapse is seen in the IAP-20 group. *p<0.001. IAP, intra-abdominal pressure.

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