Mogroside V protects against acetaminophen-induced liver injury by reducing reactive oxygen species and c-jun-N-terminal kinase activation in mice

Abstract

Background: High levels of acetaminophen (APAP) consumption can result in significant liver toxicity. Mogroside V (MV) is a bioactive, plant-derived triterpenoid known for its various pharmacological activities. However, the impact of MV on acute liver injury (ALI) is unknown.

Aim: To investigate the hepatoprotective potential of MV against liver damage caused by APAP and to examine the underlying mechanisms.

Methods: Mice were divided into three groups: Saline, APAP and APAP + MV. MV (10 mg/kg) was given intraperitoneally one hour before APAP (300 mg/kg) administration. Twenty-four hours after APAP exposure, serum transaminase levels, liver necrotic area, inflammatory responses, nitrotyrosine accumulation, and c-jun-N-terminal kinase (JNK) activation were assessed. Additionally, we analyzed reactive oxygen species (ROS) levels, JNK activation, and cell death in alpha mouse liver 12 (AML12) cells.

Results: MV pre-treatment in vivo led to a reduction in the rise of aspartate transaminase and alanine transaminase levels, mitigated liver damage, decreased nitrotyrosine accumulation, and blocked JNK phosphorylation resulting from APAP exposure, without affecting glutathione production. Similarly, MV diminished the APAP-induced increase in ROS, JNK phosphorylation, and cell death in vitro.

Conclusion: Our study suggests that MV treatment alleviates APAP-induced ALI by reducing ROS and JNK activation.

Keywords: Acetaminophen; C-jun-N-terminal kinase; Liver injury; Mogroside V; Reactive oxygen species.

Figure 1

Mogroside V attenuates acetaminophen-induced acute liver injury. A: Seven consecutive days of oral Mogroside V (MV) administration decreased serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in mice 24 hours after acetaminophen (APAP) treatment. n = 4-8 per group; B: A single dose of oral MV administration decreased serum ALT and AST levels in mice 6 and 24 hours following APAP treatment. n = 4-8 per group; C and D: A single dose of oral MV administration reduced the necrotic area and TUNEL⁺ cells in the livers of mice 12 hours after APAP exposure. n = 4 per group. Scale bar, 50 μm. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001, ^dP < 0.0001. ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; MV: Mogroside V; APAP: Acetaminophen.

Figure 2

Mogroside V reduces the infiltration of inflammatory cells in the liver of acetaminophen-treated mice. A: Representative images of immunohistochemistry (IHC) staining and the relative intensity of CD68; B: Representative images of IHC staining and the relative intensity of S100A9. Scale bar, 50 μm. n = 4-8 per group. ^aP < 0.05, ^bP < 0.01, ^dP < 0.0001. MV: Mogroside V; APAP: Acetaminophen.

Figure 3

Mogroside V has no effect on glutathione depletion in the liver following acetaminophen-induced acute liver injury. Mice were pretreated with or without Mogroside V for one hour prior to acetaminophen (APAP) administration, and glutathione levels were measured 2 and 6 hours after APAP exposure. n = 4-8 per group. MV: Mogroside V; APAP: Acetaminophen; GSH: Glutathione.

Figure 4

Mogroside V inhibits acetaminophen-induced c-jun-N-terminal kinase activation in the liver of acetaminophen-treated mice. Mice were pretreated with or without Mogroside V for one hour prior to acetaminophen administration, and the protein level of phosphorylated c-jun-N-terminal kinase (JNK) and total JNK were measured six hours later. n = 3-5 per group. ^aP < 0.05, ^bP < 0.01. MV: Mogroside V; APAP: Acetaminophen; GSH: Glutathione.

Figure 5

Mogroside V reduces nitrotyrosine levels in the liver of acetaminophen-treated mice. Representative images of immunohistochemistry staining and the corresponding relative intensity of nitrotyrosine are presented. Scale bar, 50 μm. n = 4 per group. ^bP < 0.01. MV: Mogroside V; APAP: Acetaminophen; GSH: Glutathione.

Figure 6

Mogroside V attenuates acetaminophen-induced hepatocytes damage in AML12 cells. AML 12 cells were treated with acetaminophen (APAP) (20 mM) supplemented with or without mogroside V (100 μM). A: Western blotting was used to detect the protein expression of p-c-jun-N-terminal kinase (JNK) and JNK after APAP exposure for six hours; B: Reactive oxygen species levels were detected 3 hours after APAP exposure; C: Cell death was measured by PI/Hoechst 33342 staining 24 hours after APAP exposure. Scale bar, 100 μm. MV: Mogroside V; APAP: Acetaminophen; GSH: Glutathione.