Nardostachys jatamansi inhibits severe acute pancreatitis via mitogen-activated protein kinases

Abstract

Previously, we reported that Nardostachys jatamansi (NJ) attenuated cerulein-induced mild acute pancreatitis (AP). In the present study, we investigated the ability of NJ to ameliorate severe acute pancreatitis (SAP) induced by a choline-deficient diet supplemented with ethionine (CDE). An NJ extract was orally administered ad libitum via the water during administration of the CDE. After three days, the CDE was replaced with a normal diet. After four days of normal feeding the mice were sacrificed and the blood and pancreas were obtained for further investigation. NJ treatment reduced SAP-induced pancreatic damage, as shown by histology. NJ treatment also inhibited neutrophil infiltration into the pancreas. NJ also inhibited the secretion of digestive enzymes and cytokine production, and inhibited the activation of mitogen-activated protein kinases (MAPKs) in the SAP-challenged pancreas. These data suggest that NJ protects against pancreatic injury in CDE-induced SAP by deactivating MAPKs.

Figure 1

Effect of NJ on CDE-induced SAP. (A) Magnification (×100) of representative hematoxylin and eosin (H&E)-stained pancreas sections of control mice and mice pre-treated with NJ (5 or 10 mg/ml). (B) The treatment method. (C) MPO activity was measured in the pancreas. Data are represented as the means ± SEM (n=6 in each group). ^*P<0.05 vs. control group and ^†P<0.05 vs. CDE-induced SAP mice. P<0.05 was considered to indicate a statistically significant result. The results were similar in 3 additional experiments. NJ, Nardostachys jatamansi; CDE, choline-deficient diet supplemented with ethionine; SAP, severe acute pancreatitis; MPO, myeloperoxidase.

Figure 2

Effect of NJ on the production of digestive enzymes. Serum samples were used to measure (A) amylase and (B) lipase. Data are represented as the means ± SEM (n=6 in each group). ^*P<0.05 vs. control group and ^†P<0.05 vs. CDE-induced SAP mice. P<0.05 was considered to indicate a statistically significant result. The results were similar in 3 additional experiments. NJ, Nardostachus jatamansi; CDE, choline-deficient diet supplemented with ethionine; SAP, severe acute pancreatitis.

Figure 3

Effect of NJ on TNF-α, IL-1β and IL-6. To examine production of TNF-α, IL-1β and IL-6 (A) serum levels and (B) mRNA levels in the pancreas were measured by ELISA and real-time RT-PCR, respectively. Data are represented as the means ± SEM (n=6 in each group). ^*P<0.05 vs. control group and ^†P<0.05 vs. CDE-induced SAP mice. P<0.05 was considered to indicate a statistically significant result. The results were similar in 3 additional experiments. NJ, Nardostachys jatamansi; CDE, choline-deficient diet supplemented with ethionine; TNF, tumor necrosis factor; IL, interleukin; mRNA, messenger RNA; ELISA, enzyme-linked immunosorbent assay; RT-PCR, reverse-transcription polymerase chain reaction; SAP, severe acute pancreatitis.

Figure 4

Effect of NJ on NF-κB and MAPK activation. Pancreata were obtained for western blotting to detect (A) Iκ-Bα and (B) MAPKs. Representative data from 1 of at least 3 separate experiments are shown. NJ, Nardostachys jatamansi; Iκ-Bα, inhibitory κ-Bα; MAPK, mitogen-activated protein kinase; CDE, choline-deficient diet supplemented with ethionine; (p)JNK, (phospho-)c-Jun N-terminal kinases; (p)ERK, (phospho-) extracellular signal-regulated kinases.