Genistein attenuated oxidative stress, inflammation, and apoptosis in L-arginine induced acute pancreatitis in mice

Abstract

Aim: Acute pancreatitis is a common and potentially serious condition. However, a specific treatment for this condition is still lacking. Genistein, with its anti-oxidant and anti-inflammatory effects, could possibly be used to tackle the underlying pathophysiology of acute pancreatitis. Therefore, the aim of this study was to investigate the effects of genistein on oxidative stress, inflammation, and apoptosis in acute pancreatitis induced by L-arginine in mice.

Methods: Twenty-four male ICR mice were equally divided into 4 groups: Control (Con); Acute pancreatitis (AP) group: Two doses of i.p. 350 mg/100 g body weight (BW) of L-arginine were administered 1 h apart; AP and low-dose genistein (LG) group: mice were given i.p. injection of 10 mg/kg genistein 2 h prior to L-arginine injection followed by once-daily dosing for 3 days; and AP and high-dose genistein (HG) group: mice were given 100 mg/kg genistein with the similar protocol as the LG group. Pancreatic tissue was evaluated for histopathological changes and acinar cell apoptosis, malondialdehyde (MDA) levels, immunohistochemical staining for myeloperoxidase (MPO), nuclear factor-kappa beta (NF-kB), and 4-hydroxynonenal (4-HNE). Serum levels of amylase (AMY), c-reactive protein (CRP), and interleukin (IL)-6 were measured.

Results: Significant increases in the degree of acinar cell apoptosis, pancreatic MDA, serum IL-6 and amylase, MPO, NF-kB and 4-HNE positivity were observed in the AP group. All these parameters declined after low- and high-dose genistein treatment. Severe pancreatic inflammation, edema, and acinar cell necrosis were observed in the AP group. Significant improvement of histopathological changes was seen in both low- and high-dose genistein groups. There were no significant differences in any parameters between low and high doses of genistein.

Conclusion: Genistein could attenuate the severity of histopathological changes in acute pancreatitis through its anti-oxidant, anti-inflammatory, and anti-apoptotic properties.

Keywords: Acute pancreatitis; Anti-apoptosis; Anti-inflammation; Anti-oxidant; Genistein; Mice.

Fig. 1

Effects of genistein on A body weight change, B serum amylase, C serum IL-6, D serum CRP, and E pancreatic MDA in mice with acute pancreatitis. Data are expressed as mean ± SEM. ^a1p < 0.01, ^a2p < 0.05 vs. Con group; ^b1p < 0.01, ^b2p < 0.05 vs. AP group (n = 6 mice per group). Con, Control group; AP, Acute pancreatitis group; LG, Low-dose genistein group; HG, High-dose genistein group

Fig. 2

Pancreatic histopathology of hematoxylin–eosin staining at a magnification of 400, scale bar 100 µm. A Control group showed normal pancreatic architecture without any evidence of tissue damage; B Acute pancreatitis group showed severe tissue damage characterized by neutrophil infiltration (red arrow), edema, and acinar cell necrosis; C Low-dose genistein group showed reduced neutrophil infiltration, edema, and acinar cell necrosis; D High-dose genistein group showed reduced neutrophil infiltration, edema, and acinar cell necrosis

Fig. 3

Representative images of TUNEL staining for the evaluation of apoptotic acinar cells at a magnification of 400, scale bar 100 µm. A Control group; B Acute pancreatitis group; C Low-dose genistein group; D High-dose genistein group. Red arrows indicate positive TUNEL staining cells

Fig. 4

Representative images of immunohistochemistry for MPO expression in the pancreas at a magnification of 400, scale bar 100 µm. A Control group; B Acute pancreatitis group; C Low-dose genistein group; D High-dose genistein group. Red arrows indicate positive MPO staining cells

Fig. 5

Representative images of immunohistochemistry for NF-kB expression in the pancreas at a magnification of 400, scale bar 100 µm. A Control group; B Acute pancreatitis group; C Low-dose genistein group; D High-dose genistein group. Red arrows indicate positive NF-kB staining cells

Fig. 6

Representative images of immunohistochemistry for 4-HNE expression in the pancreas at a magnification of 400, scale bar 100 µm. A Control group; B Acute pancreatitis group; C Low-dose genistein group; D High-dose genistein group. Red arrows indicate positive 4-HNE staining cells