Gardenia jasminoides protects against cerulein-induced acute pancreatitis

Abstract

Aim: To investigate the effect of Gardenia jasminoides (GJ) on cerulein-induced acute pancreatitis (AP) in mice.

Methods: C57BL/6 mice weighing 18-20 g were divided into three groups. (1) Normal saline-treated group, (2) treatment with GJ at a dose of 0.1 g/kg, (3) treatment with GJ at a dose of 1 g/kg. GJ was administered orally (n = 6 per group) for 1 wk. Three hours later, the mice were given an intraperitoneal injection of cerulein (50 microg/kg), a stable cholecystokinin (CCK) analogue, every hour for a total of 6 h as described previously. The mice were sacrificed at 6 h after completion of cerulein injections. Blood samples were obtained to determine serum amylase, lipase and cytokine levels. The pancreas was rapidly removed for morphologic examination and scoring. A portion of pancreas was stored at -70 degree and prepared for the measurement of tissue myeloperoxidase (MPO) activity, an indicator of neutrophil sequestration, and for reverse-transcriptase PCR (RT-PCR) and real-time PCR measurements.

Results: Treatment with GJ decreased significantly the severity of pancreatitis and pancreatitis-associated lung injury. Treatment with GJ attenuated the severity of AP compared with saline-treated mice, as shown by reduction in pancreatic edema, neutrophil infiltration, serum amylase and lipase levels, serum cytokine levels, and mRNA expression of multiple inflammatory mediators.

Conclusion: These results suggest that GJ attenuated the severity of AP as well as pancreatitis-associated lung injury.

Figure 1

Effects of GJ pretreatment on the (A) PW/BW, (B) serum amylase activity, and (C) serum-lipase activity in cerulein induced AP. The study groups were treated as indicated in the experimental protocol. The mean ± SE of the six animals are shown. ^aP < 0.05 vs saline treatment; ^bP < 0.05 vs cerulein treatment alone.

Figure 2

Effect of GJ on (A) TNF-α, (B) IL-1β and (C) IL-6 secretion in cerulein-induced AP. Mice were treated as indicated in the experimental protocol. The mean ± SE of six animals are shown. The study groups were treated as indicated in the experimental protocol. ^aP < 0.05 vs saline treatment; ^bP < 0.05 vs cerulein treatment alone.

Figure 3

Effect of GJ on TNF-α, IL-6 and IL-1 mRNA levels in cerulein-induced AP. The mice were sacrificed at 1, 3 and 6 h after six injections. Levels of pancreatic mRNA were quantified by real-time PCR for (A) TNF-α, (B) IL-1β, and (C) IL-6. The mean ± SE of six animals are shown. ^aP < 0.05 vs saline treatment; ^bP < 0.05 vs cerulein treatment alone.

Figure 4

Effects of the GJ on pancreatic inflammatory changes following pancreatitis. A: Representative H&E-stained sections of pancreas in control mice (CON) who were not given cerulein, in mice given cerulein (CAE), and in mice given GJ (1 mg/kg) at the same time as the first cerulein injection; B: Histological sections of pancreas harvested 12 h after injection of saline (CON), cerulein alone, or GJ (1 or 0.1 mg/kg) given at the same time as the first injection of cerulein. The results were scored from 0 (normal) to 3 (severe) for edema, inflammation, vacuolization, and necrosis. ^aP < 0.05 vs saline treatment; ^bP < 0.05 vs cerulein treatment alone. The figure shows the results of one experiment in which 4-5 mice were tested per group. The results obtained were similar to those in three additional experiments (× 200).

Figure 5

GJ reduced the severity of AP-associated lung injury. A: Representative H&E-stained sections of the pancreas in control mice (CON) not given cerulein, in mice given cerulein (CAE), and in mice given GJ (1 mg/kg) at the same time as the first cerulein injection; B: Histology sections of the lung harvested 12 h after administration of saline (CON), cerulein alone, or GJ (1 or 0.1 mg/kg) given at the same time as the first injection of cerulein. The results were scored from 0 (normal) to 3 (severe) for edema, inflammation, vacuolization, and necrosis. ^aP < 0.05 vs saline treatment; ^bP < 0.05 vs cerulein treatment alone. The figure shows one experiment in which 4-5 mice were tested per group. The results obtained were similar to those in three additional experiments (× 200).

Figure 6

MPO activity was measured in the pancreas 6 h after completion of the cerulein injections and in saline-injected control mice (CON). The data are expressed as MPO activity (U/mg protein). ^aP < 0.05 vs saline treatment; ^bP < 0.05 vs cerulein treatment alone. The figure shows the results of one experiment in which 5-6 mice were tested per group.