Effect of biologically active fraction of Nardostachys jatamansi on cerulein-induced acute pancreatitis

Figure 1 Fractionation of the aqueous extract of Nardostachys jatamansi. NJ: Nardostachys jatamansi; HPLC: High-performance liquid chromatography.

Figure 2 High-performance liquid chromatography findings of the aqueous extract of Nardostachys jatamansi (A) and the 4th fraction of Nardostachys jatamansi (B).

Figure 3 Effects of the 4th fraction of Nardostachys jatamansi on inflammatory events in the pancreas after pancreatitis. A: Representative hematoxylin-eosin stained sections of the pancreas in the control mice not administered cerulein, in mice given cerulein, and in mice given Nardostachys jatamansi (NJ) (1 mg/kg, 5 mg/kg, or 10 mg/kg) 1 h before the first cerulein injection; B: Neutrophil infiltration was assessed on the basis of myeloperoxidase activity. This figure shows representative images of 1 experiment that involved 6 mice. The results were similar in 3 additional experiments. ^aP < 0.05 vs the saline treatment; ^cP < 0.05 vs cerulein treatment alone. Original magnification: × 100. AP: Acute pancreatitis.

Figure 4 Effects of the 4th fraction of Nardostachys jatamansi on acute pancreatitis-associated lung injury. A: Representative hematoxylin-eosin stained sections of the lung in the control mice not given cerulein, in mice given cerulein, and in mice given Nardostachys jatamansi (NJ) (1 mg/kg, 5 mg/kg, or 10 mg/kg) 1 h before the first cerulein injection; B: Neutrophil infiltration was assessed on the basis of myeloperoxidase activity. This figure shows representative images from 1 experiment that involved 6 mice. The results were similar in 3 additional experiments. ^aP < 0.05 vs the saline treatment; ^cP < 0.05 vs cerulein treatment alone. Original magnification: × 100. AP: Acute pancreatitis.

Figure 5 Effects of the 4th fraction of Nardostachys jatamansi pretreatment on pancreatic weight/body weight (A), serum amylase activity (B), and serum lipase activity (C) during cerulein-induced acute pancreatitis. Mice pretreated with Nardostachys jatamansi (NJ) were challenged with intraperitoneal injections of cerulein (50 μg/kg). Mice were killed 6 h after the last cerulein injection. Their serum and pancreas were harvested and (A) pancreatic weight (PW)/body weight (BW) ratio and the levels of digestive enzymes such as (B) amylase and (C) lipase were measured as indicated in the experimental protocol. Data show the mean ± SE for 6 mice in 1 group. The results were similar in 3 additional experiments. ^aP < 0.05 vs the saline treatment; ^cP < 0.05 vs cerulein treatment alone. AP: Acute pancreatitis.

Figure 6 Effects of the 4th fraction of Nardostachys jatamansi on interleukin 1, interleukin 6, and tumor necrosis factor during cerulein-induced pancreatitis. Mice pretreated with Nardostachys jatamansi (NJ) were challenged with intraperitoneal injections of cerulein at a supramaximal dose (50 μg/kg). Mice were killed at 6 h after the last cerulein injection. Levels of serum cytokines were measured by enzyme-linked immunosorbent assay (A). Levels of pancreatic mRNA for interleukin (IL)-1, IL-6, and tumor necrosis factor (TNF)-α were quantified using real-time reverse transcriptase polymerase chain reaction (B). Data show the mean ± SE for 6 mice in 1 group. The results were similar in 3 additional experiments. ^aP < 0.05 vs the saline treatment; ^cP < 0.05 vs cerulein treatment alone. AP: Acute pancreatitis.

Figure 7 Effects of the 4th fraction of Nardostachys jatamansi on heme oxygenase-1 expression in the pancreas. Mice were treated with saline, or Nardostachys jatamansi (NJ) (10 mg/kg). Then, the pancreas was harvested at the indicated time. A: The protein level of heme oxygenase-1 (HO-1) in the pancreas was measured using Western blotting. β-actin was used as the loading control; B: The mRNA expression of HO-1 was measured using real-time reverse transcriptase polymerase chain reaction (RT-PCR). Mice were treated with the indicated dose of NJ4. At 6 h after NJ4 injection, the pancreas was harvested; C: The protein level of HO-1 in the pancreas was measured using Western blotting. β-actin was used as the loading control; D: The mRNA expression of HO-1 was measured using real-time RT-PCR. The results were similar in 3 additional experiments. ^aP < 0.05 vs the saline treatment. Cur: Curcumin.

Figure 8 Effects of the 4th fraction of Nardostachys jatamansi on heme oxygenase-1 expression in acinar cells and cerulein-induced acinar cell death. Pancreatic acinar cells were pretreated with Nardostachys jatamansi (NJ) (20 μg/mL), and then the cells were harvested at the indicated time. A, B: The protein level (A) and mRNA level (B) of heme oxygenase-1 (HO-1) in pancreatic acini were measured. Pancreatic acinar cells were pretreated with the indicated dose of NJ4. Then, the cells were harvested at 6 h; C, D: The protein level (C) and mRNA level (D) of HO-1 in the pancreatic acini were detected. The pancreatic acinar cells were pretreated with the indicated dose of NJ4 and then stimulated with cerulein (10 nmol/L); E: After 6 h of cerulein stimulation, cell viability was measured as described in the experimental protocol. Pancreatic acinar cells were pretreated with ZnPP (10 μmol/L), an HO-1 inhibitor, for 1 h and then treated with NJ4 (20 μg/mL), curcumin (10 μmol/L); F: At 1 h after treatment, cerulein (10 nmol/L) was added; After 6 h of cerulein stimulation, cell viability was measured. The results were similar in 3 additional experiments. ^aP < 0.05 vs the saline treatment; ^cP < 0.05 vs cerulein treatment alone. Cur: Curcumin.

Figure 9 Effects of Nardostachys jatamansi-2 on heme oxygenase-1 expression in acinar cells and cerulein-induced acinar cell death. Pancreatic acinar cells were pretreated with Nardostachys jatamansi (NJ4)-2 (10 μg/mL), and then the cells were harvested at the indicated time. A, B: The protein level (A) and mRNA level (B) of heme oxygenase-1 (HO-1) in pancreatic acini were measured. Pancreatic acinar cells were pretreated with the indicated dose of NJ4-2; C, D: Then, the cells were harvested at 6 h, the protein level (C) and mRNA level (D) of HO-1 in the pancreatic acini were detected. The pancreatic acinar cells were pretreated with the indicated dose of NJ4 and then stimulated with cerulein (10 nmol/L); E: After 6 h of cerulein stimulation, cell viability was measured as described in the experimental protocol. The results were similar in 3 additional experiments. ^aP < 0.05 vs the saline treatment; ^cP < 0.05 vs cerulein treatment alone.