Bees' honey attenuation of metanil-yellow-induced hepatotoxicity in rats

Abstract

The present study aims to investigate the protective effect of bees' honey against metanil-yellow-induced hepatotoxicity in rats. Rats were divided into 7 groups: control group; three groups treated with 50, 100, and 200 mg/kg metanil yellow, and three groups treated with metanil yellow plus 2.5 mg · kg(-1) · day(-1) bees' honey for 8 weeks. The obtained data showed that the antioxidant/anti-inflammatory activity of bees' honey reduced the oxidative stress in the liver tissue and downregulated the inflammatory markers. In addition, the elevated levels of AGE and the activated NF- κ B in the metanil-yellow-treated animals were significantly attenuated. Moreover, the levels of TNF- α and IL-1 β were significantly attenuated as a result of bees' honey administration. Furthermore, the histopathological examination of the liver showed that bees' honey reduced fatty degeneration, cytoplasmic vacuolization, and necrosis in metanil-yellow-treated rats. In conclusion, the obtained data suggest that bees' honey has hepatoprotective effect on acute liver injuries induced by metanil-yellow in vivo, and the results suggested that the effect of bees' honey against metanil yellow-induced liver damage is related to its antioxidant/anti-inflammatory properties which attenuate the activation of NF- κ B and its controlled genes like TNF- α and IL-1 β .

Figure 1

Binding activity of NF-κB-p65 (a). Group 1, untreated (normal control); groups 2–4, rats treated with 50, 100, 200 mg/kg metanil yellow, respectively; and groups 5–7, rats were treated with metanil yellow as in groups 2–4 plus bees' honey 2.5 mg·kg⁻¹d⁻¹ for 8 weeks. Binding activity of NF-κB-p65 to its consensus sequence was assayed by EMSA of total protein extracts. Quantification of activated NF-κB-p65 was performed by densitometric analysis of relative EMSA band intensities (b) using the normal control band as a reference band. All values expressed as mean ± S.D. of triplicate tests (n = 10). ^a P < 0.05 versus group1, ^b P < 0.05 versus group 2, ^c P < 0.05 versus group 3, ^d P < 0.05 versus group 4.

Figure 2

Effect of bees' honey on nitric oxide levels of rat liver homogenate treated with metanil yellow. Group 1, control group; groups 2–4, rats treated with 50, 100, and 200 mg metanil yellow/kg body weight for 8 weeks, respectively; groups 5–7, rat groups treated with 50, 100, and 200 mg metanil yellow/kg body weight plus 2.5 g/kg bees' honey daily for 8 weeks, respectively. All values were expressed as mean ± S.D. of triplicate tests (n = 10). ^a P < 0.05 versus group1, ^b P < 0.05 versus group 2, ^c P < 0.05 versus group 3, ^d P < 0.05 versus group 4.

Figure 3

Effect of bees' honey on TNF-α level of rat liver treated with metanil yellow. Group 1, control group; groups 2–4, rats treated with 50, 100, and 200 mg metanil yellow/kg body weight for 8 weeks, respectively; groups 5–7, rat groups treated with 50, 100, and 200 mg metanil yellow/kg body weight plus 2.5 g/kg bees' honey daily for 8 weeks, respectively. All values were expressed as mean ± S.D. of triplicate tests (n = 10). ^a P < 0.05 versus group1, ^b P < 0.05 versus group 2, ^c P < 0.05 versus group 3, ^d P < 0.05 versus group 4.

Figure 4

Effect of bees' honey on IL-1β levels of rat liver treated with metanil yellow. Group 1, control group; groups 2–4, rats treated with 50, 100, and 200 mg metanil yellow/kg body weight for 8 weeks, respectively; groups 5–7, rat groups treated with 50, 100, and 200 mg metanil yellow/kg body weight plus 2.5 g/kg bees honey daily for 8 weeks, respectively. All values were expressed as mean ± S.D. of triplicate tests (n = 10). ^a P < 0.05 versus group1, ^b P < 0.05 versus group 2, ^c P < 0.05 versus group 3, ^d P < 0.05 versus group 4.

Figure 5

Histopathological findings of rat liver from different groups. Livers were removed, fixed, and embedded in paraffin. Sections were stained with hematoxylin-eosin (×200). (a) control group; (b)–(d) rats treated with 50, 100, and 200 mg metanil yellow/kg body weight for 8 weeks, respectively; (e)–(g) rat groups treated with 50, 100, and 200 mg metanil yellow/kg body weight plus 2.5 g/kg bees' honey daily for 8 weeks, respectively.