Efficiency of Hesperidin against Liver Fibrosis Induced by Bile Duct Ligation in Rats

Abstract

Introduction: Bile duct ligation (BDL) and subsequent cholestasis are associated with oxidative stress and liver injury and fibrosis. Hesperidin (3,5,7-trihydroxyflavanone 7-rhamnoglucoside) is a flavanone glycoside abundant in citrus fruits. It has positive effects on diabetic retinopathy, reduced platelet aggregation, and blood flow alterations and has the potential to reduce liver injury in oxidative stress. The aim of this study was to evaluate the hepatoprotective effects of hesperidin on BDL-induced liver injury in rats.

Methods: A total of 48 adult male Wistar rats were equally allocated to six eight-rat groups, namely, a healthy group, a sham group, a BDL+Vehicle group (BDL plus treatment with distilled water), a BDL+VitC group (BDL plus treatment with vitamin C 4.25 μg/kg), a BDL+Hesp100 group (BDL plus treatment with hesperidin 100 mg/kg/day), and a BDL+Hesp200 group (BDL plus treatment with hesperidin 200 mg/kg/day). Treatments were orally provided for 21 consecutive days. Finally, rats were sacrificed through heart blood sampling. Blood samples were centrifuged, and liver function, oxidative stress, and antioxidant parameters were assessed. Liver tissue was also assessed for oxidative stress, antioxidant, and histological parameters. The expression of inflammatory genes, namely, TGFβ1, iNOS, Caspase-3, and α-SMA, was measured through polymerase chain reaction. Findings. Hesperidin supplementation was associated with significant decrease in the levels of liver enzymes, bilirubin, nitric oxide, malondialdehyde, protein carbonyl, and inflammatory gene expression; significant increase in the levels of total antioxidant capacity, glutathione, and superoxide dismutase and catalase enzyme activity; and significant improvement in the histological morphology and structure of the liver parenchyma.

Conclusion: Hesperidin has significant positive effects on liver morphology and structure, inflammation, fibrosis, and oxidative stress in rats with BDL-induced cholestatic liver injury.

Figure 1

Effects of hesperidin on the histopathological changes of the liver in BDL-afflicted rats. Representative photomicrographs of liver sections processed for H and E staining (10; scale bar 5 mm). Sham ×400 (a); BDL-control ×100 (b); BDL+VitC ×100 (c); BDL+Hesp100 ×100 (d); BDL+Hesp100 ×400 (e); and BDL+Hesp200 ×100 (f). (a) represents the normal liver histopathology. (e) is a section of (d) at ×400 magnification. In (b)–(e), bile duct hyperplasia is evident. All these lesions markedly decreased in rats treated with 200 mg/kg hesperidin.

Figure 2

The levels of nitric oxide, malondialdehyde (MDA), and protein carbonyl in liver tissue and serum in different groups. Data are presented as mean ± SD. SC: sham control; BDL: bile duct ligation; VitC: vitamin C; Hesp: hesperidin. ^#Significant difference with the sham group. ^∗Significant difference with the BDL+Vehicle group. ^{^}Significant difference with the BDL+VitC group.

Figure 3

The levels of total antioxidant capacity (TAC) and glutathione (GSH) in liver tissue and serum in different groups. Data are presented as mean ± SD. SC: sham control; BDL: bile duct ligation; VitC: vitamin C; Hesp: hesperidin. ^#Significant difference with the sham group. ^∗Significant difference with the BDL+Vehicle group. ^{^}Significant difference with the BDL+VitC group.

Figure 4

The levels of superoxide dismutase (SOD) and catalase enzyme activity in liver tissue and serum in different groups. Data are presented as mean ± SD. SC: sham control; BDL: bile duct ligation; VitC: vitamin C; Hesp: hesperidin. ^#Significant difference with the sham group. ^∗Significant difference with the BDL+Vehicle group. ^{^}Significant difference with the BDL+VitC group.

Figure 5

The level of the β-actin, TGFβ1, iNOS, Caspase-3, and α-SMA gene expression in different groups. Data are presented as mean ± SD. SC: sham control; BDL: bile duct ligation; VitC: vitamin C; Hesp: hesperidin; TGFβ1: transforming growth factor β1; iNOS: inducible nitric oxide synthase; α-SMA: alpha smooth muscle actin. ^#Significant difference with the sham group. ^∗Significant difference with the BDL+Vehicle group. ^{^}Significant difference with the BDL+VitC group.