Histopathological and biochemical assessment of d -limonene-induced liver injury in rats

Abstract

The aim of the present work was to develop a biochemical, histologic and immunohistochemical study about the potential hepatotoxic effect of d-limonene – a component of volatile oils extracted from citrus plants. Blood alkaline phosphatase (ALP), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) from d-limonene-treated animals were determined and compared to morphologic hepatic lesions in order to investigate the possible physiopathologic mechanisms involved in the liver toxicity, in experimental animals treated with d-limonene. Wistar rats were randomly divided into seven groups: two control groups (untreated or receiving only vehicle, tween-80); one positive control (vehicle); two experimental groups treated with d-limonene at doses of 25 mg/kg/day and 75 mg/kg/day for 45 days, and two other groups treated with the same doses for 30 days and kept under observation during 30 more days. Biochemical data showed significant reduction in ALT levels in the animals treated with 75 mg/kg of d-limonene. Histological analysis revealed some hepatocyte morphological lesions, including hydropic degeneration, microvesicular steatosis and necrosis, Kupffer cell hyperplasia and incipient fibrosis. By immunohistochemistry, influx of T (CD3+) and cytotoxic (CD8+) lymphocytes was observed in the rats treated with d-limonene at both dose levels. In conclusion, it is possible that d-limonene has been directly responsible for hepatic parenchymal and matrix damage following subchronic treatment with d-limonene.

Peer Review #3 of "Sunset Yellow and Allura Red modulate Bcl2 and COX2 expression levels and confer oxidative stress-mediated renal and hepatic toxicity in male rats (v0.2)"

“…The measurement of antioxidant capacity is an appropriate indication for the total antioxidant defences of various kinds of tissues and organs (Ferrara, Gerber & LeCouter, 2003). Our study showed a reduced antioxidant capacity in rats treated with the dyes that can be attributed to the accelerated production of free radicals during (Kosif et al, 2010;Ramos et al, 2015). Meanwhile, the marked increase in infiltration of leucocytes implies a possible inflammatory response in affected tissues, as noted in earlier works (Soltan & Shehata, 2012;El-Desoky et al, 2017) following exposure to synthetic colouring agents in foods.…”

Sunset Yellow and Allura Red modulate Bcl2 and COX2 expression levels and confer oxidative stress-mediated renal and hepatic toxicity in male rats

“…Inflammation, necrosis and vacuolisation of the tubular epithelium occurred, in addition to the destruction of glomeruli, with thickening of Bowman’s capsule. The hydropic degeneration of hepatocytes and vacuolisation of tubular cells indicate the occurrence of intracellular oedema, as a result of toxicity or immune aggressions ( Kosif et al, 2010 ; Ramos et al, 2015 ). Meanwhile, the marked increase in infiltration of leucocytes implies a possible inflammatory response in affected tissues, as noted in earlier works ( Soltan & Shehata, 2012 ; El-Desoky et al, 2017 ) following exposure to synthetic colouring agents in foods.…”

D‐limonene: A multifunctional compound with potent therapeutic effects

“…D‐limonene intake and/or exposure (via cosmetic products) for a very long time, especially at higher doses may show toxicity (Ravichandran et al., 2018). Some preclinical studies have also reported the renal (Hard & Whysner, 1994) and liver toxicity of D‐limonene (Ramos et al., 2015).…”