Acute pancreatitis is usually diagnosed by the presence of acute abdominal pain and increased serum amylase and lipase levels. Acute pancreatitis may cause damage to other organs because of systemic inflammation and cytokines. In particular, severe acute pancreatitis (SAP) causes other organ damage and has a high mortality rate (range, 15%–40%) [
Because of these inflammatory cytokines, SAP causes systemic inflammation that results in damage to other organs, such as the lung, liver, kidney, and stomach [
We obtained approval from the local ethical committee of the Research and Application Center for Experimental Animals at our university. A total of 58 male BALB-C mice (25 g) were divided into four control and three treatment groups. The control groups were group 1, saline (6 mice); group 2, acute pancreatitis (with intraperitoneal caerulein; 9 mice); group 3, octreotide (6 mice); and group 4, diclofenac sodium (6 mice). The treatment groups were group 5, octreotide with acute pancreatitis (9 mice); group 6, diclofenac sodium with acute pancreatitis (10 mice); and group 7, octreotide plus diclofenac sodium with acute pancreatitis (12 mice). The animals were not used in another study before our model, given standard laboratory foods and drinking water, and housed with a 12 : 12 h light/dark cycle at 21°C–24°C. The animals received only water for 12 h before the beginning of the experiment. All experiments were performed under anesthesia, and the mice were scarified 6 h after the last injection.
The anesthesia protocol consisted of 2.5 mg xylazine plus 7.5 mg ketamine per 100 g body weight in 200
All tissues were fixed in 10% formaldehyde solution for 24 h, processed using the autotechnicon instrument (Leica ASP300; Leica Biosystems, Wetzlar, Germany), and embedded in paraffin blocks. A microtome was used to cut 5
H&E-stained preparations were evaluated by a single experienced pathologist in a blinded fashion using an Olympus BX51 light microscope. Histopathologic criteria were evaluated for the kidney (edema, congestion, intraparenchymal inflammation, perirenal fat tissue inflammation, tubular injury, and tubular stasis), lung (edema, congestion, neutrophil infiltration, mononuclear infiltration, alveolary wall thickness, and emphysematous changes), stomach (edema, congestion, neutrophil infiltration, mononuclear infiltration, and erosion), and liver (congestion, hydropic degeneration, necrosis, portal inflammation, and portal fibrosis). Scores were graded as grades 0 (no pathology), 1 (mild), 2 (moderate), and 3 (severe).
Data were analyzed using IBM SPSS Statistics version 17.0 software (IBM Corporation, Armonk, NY, USA). Descriptive statistics are shown as the median (25th–75th) percentile for ordinal data. Statistical significance in the differences in histopathologic scores among the groups was evaluated with the Kruskal–Wallis test. When
Histopathologic investigation showed statistically significant differences between the groups in mean congestion, edema, tubular injury, perirenal fat tissue inflammation, and tubular stasis scores in kidney tissue (
Histopathological results obtained from the kidney tissue according to groups.
Group 1 ( |
Group 2 ( |
Group 3 ( |
Group 4 ( |
Group 5 ( |
Group 6 ( |
Group 7 ( |
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Congestion | 0 (0–1) | 2 (1–2.5)a | 0 (0–1)b | 0.5 (0–1)b | 1 (1–2)a,c,d | 1 (1–2)a,c,d | 1.5 (1–2)a,c,d | |
Edema | 0 (0–1) | 1 (1–2.5)a | 0 (0–0.25)b | 0 (0–0)b | 1 (1–2)a,c,d | 1 (1–2)a,c,d | 1 (1–2)a,c,d | |
Tubular injury | 0 (0–0) | 1 (1–2)a | 0 (0–0.25)b | 0 (0–1)b | 1 (1–2)a,c,d | 1 (1–2)a,c,d | 1 (1–2)a,c,d | |
Parenchymal inflammation | 0 (0–0) | 0 (0–0) | 0 (0–0) | 0 (0–0) | 0 (0–0) | 0 (0–0) | 0 (0–0) | 0.253 |
Perirenal fat tissue inflammation | 0 (0–0) | 2 (1.5–3)a | 0 (0–0)b | 0 (0–0)b | 2 (1–2)a,c,d | 1.5 (1–2)a,c,d | 1 (1–2)a,c,d | |
Tubular stasis | 0 (0–0) | 1 (0–1.5)a | 0 (0–0)b | 0 (0–0)b | 1 (0–1)a,c,d | 0 (0–1) | 0 (0–1) |
Data were shown as median (25th–75th) percentiles, the Kruskal–Wallis test. aThe difference between the considered group and group 1 was found as statistically significant (
Histopathological results obtained from the lung tissue according to groups.
Group 1 ( |
Group 2 ( |
Group 3 ( |
Group 4 ( |
Group 5 ( |
Group 6 ( |
Group 7 ( |
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Congestion | 0 (0–0.25) | 2 (2–2)a | 1 (0–1.25)b | 1 (1–2)a,b | 2 (1–2)a,c | 1 (1–1)a,b,d | 1 (1–2)a,b | |
Edema | 0 (0–1) | 1 (1–1)a | 0 (0–1)b | 0 (0–1)b | 1 (0–1) | 0 (0–0)b,d | 1 (1–1)a,c,e,f | |
Neutrophil infiltration | 0 (0–0) | 0 (0–1) | 0 (0–0) | 0 (0–0) | 0 (0–0) | 0 (0–0) | 0 (0–0)b | |
Mononuclear infiltration | 0 (0–0) | 0 (0–1.5) | 0 (0–0) | 0 (0–0) | 0 (0–0.5) | 0 (0–0)b | 0 (0–0) | |
Alveolary wall thickness | 0 (0–0) | 0 (0–0) | 0 (0–0) | 0 (0–0) | 0 (0–0) | 0 (0–0) | 0 (0–0) | >0.999 |
Emphysematous changes | 0 (0–1) | 1 (1–1)a | 0 (0–0)b | 1 (0.75–1)c | 1 (1–1)a,c | 1 (1–1)a,c | 1 (1–1)a,c |
Data were shown as median (25th–75th) percentiles, the Kruskal–Wallis test. aThe difference between the considered group and group 1 was found as statistically significant (
Histopathological results obtained from the stomach tissue according to groups.
Group 1 ( |
Group 2 ( |
Group 3 ( |
Group 4 ( |
Group 5 ( |
Group 6 ( |
Group 7 ( |
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Congestion | 0 (0–0.25) | 1 (1–1.5)a | 0 (0–1)b | 1 (0–1) | 1 (0–1)a | 1 (1–1.25)a,c | 1 (0.25–1)a | |
Edema | 0 (0–1) | 1 (1–1.5)a | 0 (0–1)b | 0.5 (0–1) | 1 (0.5–1) | 1 (1–1.25)a,c,d | 1 (0.25–1) | |
Neutropil infiltration | 0 (0–0.25) | 1 (1–2)a | 0 (0–0)b | 0 (0–0)b | 0 (0–1)b | 0 (0–1)b | 1 (1–1)a,c,d,e,f | |
Erosion | 0 (0–0) | 0 (0–0) | 0 (0–0) | 0 (0–0) | 0 (0–0) | 0 (0–0) | 0 (0–0) | >0.999 |
Mononuclear infiltration | 0.5 (0–1) | 1 (0.5–1) | 0.5 (0–1) | 1 (0–1) | 1 (0–1.5) | 1 (1–1) | 1 (1–1) | 0.272 |
Data were shown as median (25th–75th) percentiles, the Kruskal–Wallis test. aThe difference between the considered group and group 1 was found as statistically significant (
Histopathological results obtained from the liver tissue according to groups.
Group 1 ( |
Group 2 ( |
Group 3 ( |
Group 4 ( |
Group 5 ( |
Group 6 ( |
Group 7 ( |
||
---|---|---|---|---|---|---|---|---|
Congestion | 0 (0–1) | 1 (1–1.5)a | 0 (0–0.25)b | 0 (0–1)b | 1 (0–1)b | 1 (0–1)b | 1 (0–1)b | |
Hydropic degeneration | 0 (0–1) | 2 (1–2)a | 0 (0–0.25)b | 0 (0–1)b | 1 (0–1.5)b,c | 1 (0.75–1.25)a,c,d | 1 (0–1)b | |
Focal necrosis | 0 (0–0) | 0 (0–1) | 0 (0–0.25) | 0 (0–0) | 0 (0–0.5) | 0 (0–0.25) | 0 (0–0) | 0.640 |
Portal inflammation | 0 (0–0) | 0 (0–1) | 0 (0–0) | 0 (0–0) | 0 (0–0.5) | 0 (0–1) | 0 (0–0) | 0.259 |
Portal fibrosis | 0 (0–0) | 0 (0–0) | 0 (0–0) | 0 (0–0) | 0 (0–0) | 0 (0–0) | 0 (0–0) | >0.999 |
Data were shown as median (25th–75th) percentiles, the Kruskal–Wallis test. aThe difference between the considered group and group 1 was found as statistically significant (
Conover’s multiple comparison test after the Kruskal–Wallis test in multiple comparisons revealed statistically significant differences in mean congestion scores between group 1 and groups 2 and 5–7 (
There were significant differences between group 2 and groups 1, 3, and 4 (
There were significant differences between group 1 and groups 2 and 5–7 (
Significant differences were found between group 1 and groups 2 and 5–7 (
Comparing group 1 and the other groups showed significant differences between group 1 and groups 2 and 5 (
Significant differences were found between group 2 and groups 1, 3, 4, 6, and 7 (
There were statistically significant differences between group 2 and groups 1, 3, 4, and 6 (
A significant difference was found only between groups 2 and 7 (
There was no significant difference between group 1 and all other groups (
There were statistically significant differences between group 1 and groups 2 and 5–7 (
There were significant differences between group 1 and groups 2 and 5–7 (
Significant differences were noted between group 1 and groups 2 and 6 (
Statistically significant differences were noted between group 2 and groups 1 and 3–6 (
Significant differences were noted between groups 1 and 2 (
There were significant differences between group 2 and groups 3–5 and 7 (
Acute pancreatitis is a common inflammatory condition of the pancreas and begins as local inflammation. Spread of the inflammation can lead to systemic multiple organ dysfunction and death [
A previous study suggested that N-acetylcysteine treatment ameliorated pulmonary and renal tissue damages in all microscopic parameters [
Heparin administration has been suggested to have a positive influence on lung, liver, kidney, colon, and stomach microcirculatory disturbances accompanying experimental caerulin-induced acute pancreatitis [
In our study, lung congestion scores in groups 6 (diclofenac sodium treatment) and 7 (diclofenac sodium plus octreotide combination) were significantly lower than those in group 2 (acute pancreatitis). Group 6 also showed significantly lower lung edema scores than group 2. Therefore, diclofenac sodium as an anti-inflammatory drug ameliorated lung injury in experimental acute pancreatitis. Neutrophil inflammation of the lung in group 7 was significantly lower than that in group 2 in our study. Also, mononuclear inflammation of the lung was only lower in group 6 than in group 2.
Dobosz et al. [
Our study showed that congestion and edema scores for the stomach in all treatment groups were similar to those of group 2. Although neutrophil inflammation of the stomach in group 7 was similar to that in group 2, it was significantly higher than those in the other treatment groups (groups 5 and 6). We concluded that there might be a counteraction between diclofenac sodium and octreotide.
A previous study showed that gadolinium chloride reduced hepatic or systemic cytokine levels and lung injury in experimental pancreatitis [
In our study, liver congestion in all treatment groups was significantly lower than that in the acute pancreatitis group. In terms of hydropic degeneration of the liver, the diclofenac sodium treatment group was similar to the acute pancreatitis group and had significantly higher levels than the other treatment groups. However, the octreotide and combination treatment groups showed significantly lower levels than the acute pancreatitis group in terms of hydropic degeneration of the liver. The prognosis of acute pancreatitis is associated with extrapancreatic organ injuries. Also, the treatment of this situation is not clear. We showed the effects of diclofenac sodium, octreotide, and their combination on extrapancreatic organ injuries (kidney, lung, stomach, and liver) in caerulein-induced acute pancreatitis in mice. Our study demonstrated that diclofenac sodium improved experimental acute pancreatitis–associated lung edema. Diclofenac sodium may ameliorate the prognosis of acute pancreatitis by reducing lung edema.
In conclusion, diclofenac sodium is a well-known drug that may be a good choice for extrapancreatic organ injuries in cases of acute pancreatitis. Diclofenac sodium improved lung edema–associated experimental acute pancreatitis in our study. Therefore, diclofenac sodium may improve the prognosis of acute pancreatitis by reducing lung edema. Further studies are needed for clarity.
The data used to support the findings of this study are available from the corresponding author upon request.
The authors declare that they have no conflicts of interest.