Ameliorative effects of umbelliferone against acetaminophen-induced hepatic oxidative stress and inflammation in mice

Abstract

Background and purpose: Acetaminophen (APAP) is a commonly used antipyretic and pain reliever that its overdose causes acute liver toxicity. Umbelliferone (UMB) has many pharmacological effects. In this study, the hepatoprotective effect of UMB on acute hepatotoxicity induced by APAP was investigated.

Experimental approach: Forty-nine male mice were separated into seven groups. The control received vehicle (i.p.), UMB group received UMB (120 mg/kg, i.p.), APAP group was treated with a single dose of APAP (350 mg/kg, i.p.), and pretreated groups received N-acetylcysteine (NAC, 200 mg/kg, i.p.) or different doses of UMB (30, 60, and 120 mg/kg, i.p.), respectively before APAP. Twenty-four hours after APAP injection, mice were sacrificed and blood and liver samples were collected. Then, serum and tissue samples were investigated for biochemical and histological studies.

Findings/results: A single dose of APAP caused elevation in the serum liver enzymes, including alanine aminotransferase, aspartate transaminase, and alkaline phosphatase. The amounts of thiobarbituric acid reactive substances, tumor necrosis factor-alpha, and nitric oxide increased in the mice's liver tissue. Moreover, the amount of total thiol and the activity of antioxidant enzymes (catalase, superoxide dismutase, and glutathione peroxidase) significantly diminished in the APAP group. Histological results confirmed the hepatotoxicity induced by APAP. However, UMB (more effective at 60 and 120 mg/kg) lessened APAP-induced hepatic injuries, which is comparable with NAC effects.

Conclusion and implications: The findings of this study provided evidence that UMB ameliorates liver injury induced by APAP through its antioxidant and anti-inflammatory effects.

Keywords: Acetaminophen; Hepatotoxicity; Inflammation; Oxidative stress; Umbelliferone..

Fig. 1.

Effect of UMB on serum levels of (A) ALT, (B) AST, and (C) ALP in the APAP (350 mg/kg) model of hepatotoxicity in mice. The data are expressed as mean ± SEM. ^*P < 0.05 and ^***P < 0.001 indicate significant differences in comparison with the control group; ^##P < 0.01 and ^###P < 0.001 versus the APAP group. UMB, Umbelliferone; ALT, alanine aminotransferase; AST, aspartate transaminase; ALP, alkaline phosphatase; APAP, acetaminophen; NAC, N-acetylcysteine.

Fig. 2.

Effect of UMB on (A) total thiol and (B) TBARS in the APAP (350 mg/kg) model of hepatotoxicity in mice. The data are expressed as mean ± SEM. ^***P < 0.001 Indicates significant differences in comparison with the control group; ^#P < 0.05, ^##P < 0.01, and ^###P < 0.001 versus the APAP group. UMB, Umbelliferone; TBARS, thiobarbituric acid reactive substances; APAP, acetaminophen; NAC, N-acetylcysteine.

Fig. 3.

Effect of UMB on (A) CAT, (B) SOD, and (C) GPx enzyme activity in the APAP (350 mg/kg) model of hepatotoxicity in mice. The data are expressed as mean ± SEM. ^*P < 0.05, ^**P < 0.01, and ^***P < 0.001 indicate significant differences in comparison with the control group; ^#P < 0.05, ^##P < 0.01, and ^###P < 0.001 versus the APAP group. UMB, Umbelliferone; CAT, catalase; SOD, superoxide dismutase; GPx, glutathione peroxidase; APAP, acetaminophen; NAC, N-acetylcysteine.

Fig. 4.

Effect of UMB on (A) TNF-α and (B) NO in the APAP (350 mg/kg) model of hepatotoxicity in mice. The data are expressed as mean ± SEM. ^***P < 0.001 Indicates significant differences in comparison with the control group; ^#P < 0.05, ^##P < 0.01, and ^###P < 0.001 versus the APAP group. UMB, Umbelliferone; TNF-α, tumor necrosis factor alpha; NO, nitric oxide; APAP, acetaminophen; NAC, N-acetylcysteine.

Fig. 5.

Evaluation of the effect of UMB (mg/kg) on liver tissue changes in the APAP (350 mg/kg) model of hepatotoxicity in mice using hematoxylin and eosin staining; magnification: ×300, scale bar: 100 μm. Green arrows: necrosis, black arrows: infiltration of inflammatory cells, white arrows: degeneration of hepatocytes, and blue arrows: dilation of sinusoids. UMB, Umbelliferone; APAP, acetaminophen; NAC, N-acetylcysteine.

Fig. 6.

The hepatoprotective effects of umbelliferone on acute hepatotoxicity caused by APAP in mice. APAP, acetaminophen; NAPQI, N-acetyl-p-benzoquinoneimine; ALT, alanine aminotransferase; AST, aspartate transaminase; ALP, alkaline phosphatase; TBARS, thiobarbituric acid reactive substances; CAT, catalase; SOD, superoxide dismutase; GPx, glutathione peroxidase; NO, nitric oxide; TNF-α, tumor necrosis factor-alpha.