Korean J Pain.  2019 Jul;32(3):168-177. 10.3344/kjp.2019.32.3.168.

Diverse characters of Brennan’s paw incision model regarding certain parameters in the rat

• Kumar R
• Gupta S
• Gautam M
• Jhajhria S
• Ray SB

• KMID: 2455612
• DOI: http://doi.org/10.3344/kjp.2019.32.3.168

Abstract

BACKGROUND
Brennan's rodent paw incision model has been extensively used for understanding mechanisms underlying postoperative pain in humans. However, alterations of physiological parameters like blood pressure and heart rate, or even feeding and drinking patterns after the incision have not been documented as yet. Moreover, though eicosanoids like prostaglandins and leukotrienes contribute to inflammation, tissue levels of these inflammatory mediators have never been studied. This work further investigates the antinociceptive effect of protein C after intra-wound administration.
METHODS
Separate groups of Sprague-Dawley rats were used for quantitation of cyclooxygenase (COX) activity and leukotriene B4 level by enzyme-linked immunosorbent assay, as well as estimation of cardiovascular parameters and feeding and drinking behavior after paw incision. In the next part, rats were subjected to incision and 10 μg of protein C was locally administered by a micropipette. Both evoked and non-evoked pain parameters were then estimated.
RESULTS
COX, particularly COX-2 activity and leukotriene B4 levels increased after incision. Hemodynamic parameters were normal. Feeding and drinking were affected on days 1 and 3, and on day 1, respectively. Protein C attenuated non-evoked pain behavior alone up to day 2.
CONCLUSIONS
Based upon current observations, Brennan's rodent paw incision model appears to exhibit a prolonged period of nociception similar to that after surgery, with minimal interference of physiological parameters. Protein C, which is likely converted to activated protein C in the wound, attenuated the guarding score, which probably represents pain at rest after surgery in humans.

Keyword

Blood Pressure; Eicosanoids; Heart Rate; Inflammation; Nociception; Pain, Postoperative; Protein C; Rats; Wounds and Injuries

MeSH Terms

Animals
Blood Pressure
Drinking
Drinking Behavior
Eicosanoids
Enzyme-Linked Immunosorbent Assay
Heart Rate
Hemodynamics
Humans
Inflammation
Leukotriene B4
Leukotrienes
Nociception
Pain, Postoperative
Prostaglandin-Endoperoxide Synthases
Prostaglandins
Protein C
Rats*
Rodentia
Wounds and Injuries
Eicosanoids
Leukotriene B4
Leukotrienes
Prostaglandin-Endoperoxide Synthases
Prostaglandins
Protein C

Figure

• 

  Fig. 1 Activated protein C (10 μg) was administered by intra-wound route using a sterile micropipette, only once in each rat during the experiment.

• 

  Fig. 2 Estimation of peroxidase component of cyclooxygenase (COX) enzymes by calorimetric method. Control animals (n = 4) were sacrificed, before (basal) and at specific time points after incision (between 2 hr and day 3) for estimating COX activity. Maximum increase was noted at 8 hours, when COX-2 activity was greater than COX-1. Pooled tissues (n = 4) was evaluated at each time point and thus statistical analysis could not be performed.

• 

  Fig. 3 Biochemical estimation of leukotriene B4 (LTB4) by competitive ELISA in control group of animals before (basal) and after incision (between 2 hr and day 3). Maximum increase was observed at 8 hours compared to basal level. The increase persisted up to day 3. Values (n = 4) are represented as mean ± standard error of mean. *P < 0.05, **P < 0.01, ***P < 0.001.

• 

  Fig. 4 Estimation of hemodynamic parameters in control group of animals, before (baseline) and after incision (between 2 hr and day 5). Blood pressure (A) and heart rate (B) were not affected compared to baseline data. Values are expressed as mean ± standard error of mean. n = 6 at each time point.

• 

  Fig. 5 Assessment of metabolic function in control animals. Time of incision is indicated (line arrow). Feeding behaviour was affected at days 1 and 3 (A) and drinking on day 1 (B) after incision. All values were compared to baseline intake. Values are represented as mean ± standard error of mean. Food consumption was expressed as g/day. n = 6 at each time point. *P < 0.05, **P < 0.01, ***P < 0.001.

• 

  Fig. 6 Intra-wound administration of protein C (10 μg) only once, resulted in a decrease in guarding score (A) but did not affect mechanical allodynia (B) or thermal hyperalgesia (C). Treatment with activated protein C reduced guarding score from 2 hours to day 2 in comparison to control group. Values (n = 6) are represented as mean ± standard error of mean. Day 1 to 7 refers to days 1 to 7 (***P < 0.001).

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