Antioxidant and hepatoprotective effects of Artemisia dracunculus against CCl4-induced hepatotoxicity in rats

Abstract

Objective: The present study was conducted to investigate the antioxidant and hepatoprotective activity of the hydro-alcoholic extract of aerial parts of Artemisiadracunculus (HAAD) against CCl₄-induced hepatotoxicity in rats.

Materials and methods: The antioxidant activity was evaluated by reducing power, 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and 2, 20-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays. Rats were pre-treated with either 50, 100, and 200 mg/kg of HAAD or silymarin (100 mg/kg; served as the positive control group) for 15 days and they received a single dose of CCl₄ on the last day. Hepatoprotective effects were investigated by assessment of serum biochemical enzymes such as alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), total protein (TP), total bilirubin (TB), malondialdehyde (MDA), and antioxidant enzymes (SOD, CAT, GST and GSH), along with histopathological studies.

Results: Total phenolic content was 197.22±3.73 mg gallic acid equivalent/g HAAD dry weight. HAAD indicated powerful activity in FRAP, DPPH and ABTS tests. Acute toxicity study showed that the extract had an LD₅₀ of >5000 mg/kg. Oral treatment with HAAD exhibited a significant decrease in the levels of AST, ALT, ALP and TB and an increase in the level of TP. The extract significantly diminished MDA levels. The activities of the antioxidant enzymes were significantly augmented in rats pretreated with HAAD 200 mg/kg. Histopathological examination demonstrated lower liver damage in HAAD-treated groups as compared to CCl₄ groups.

Conclusion: Our findings indicated hepatoprotective effects of the hydro-alcoholic extract of A. dracunculus on CCl₄-induced hepatic damage in rats and suggested that these effects may be produced by reducing oxidative stress.

Keywords: Antioxidant; Artemisia dracunculus; Carbon Tetrachloride; Hepatotoxicity; Oxidative stress; Rats.

Figure 1

In vitro antioxidant activity of HAAD and ascorbic acid. Graph showing ferric reducing antioxidant power activity. Values are presented as mean ± SEM. HAAD: hydro-alcoholic extract of aerial parts of A. dracunculus

Figure 2

. In vitro antioxidant activity of HAAD and ascorbic acid. Graph showing DPPH radical scavenging activity. Values are presented as mean ± SEM. HAAD: hydro-alcoholic extract from aerial parts of A. dracunculus

Figure 3

In vitro antioxidant activity of HAAD and ascorbic acid. Graph showing ABTS radical scavenging activity. Values are presented as mean ± SEM. HAAD: hydro-alcoholic extract of aerial parts of A. dracunculus

Figure 4

Effect of HAAD and silymarin on ALT, AST, and ALP activity in CCl₄-induced hepatotoxicity. Groups – I: normal saline; II: CCl₄ (CCl₄ group); III: Silymarin (100 mg/kg) + CCl₄ (10 ml/kg); IV: HAAD (50 mg/kg) + CCl₄ (10 ml/kg); V: HAAD (100 mg/kg) + CCl₄ (10 ml/kg); VI: HAAD (200 mg/kg) + CCl₄ (10 ml/kg). Values are mean ± S.E.M. (n=6). ^###p<0.001 vs. group (I); ^*p<0.05, ^**p<0.01, and ^***p<0.001 vs. CCl₄ group (II).

Figure 5

Effect of HAAD and silymarin on total bilirubin (TB) in CCl₄-induced hepatotoxicity. Groups – I: normal saline; II: CCl₄ (CCl₄ group); III: Silymarin (100 mg/kg) + CCl₄ (10 ml/kg); IV: HAAD (50 mg/kg) + CCl₄ (10 ml/kg); V: HAAD (100 mg/kg) + CCl₄ (10 ml/kg); VI: HAAD (200 mg/kg) + CCl₄ (10 ml/kg). Values are mean ± S.E.M. (n=6). ^##p<0.01 vs. group (I); ^*p<0.05, ^**p<0.01, and ^***p<0.001 vs. CCl₄ group (II

Figure 6

Effect of HAAD and silymarin on total protein (TP) in CCl₄-induced hepatotoxicity. Groups – I: normal saline; II: CCl4 (CCl₄ group); III: Silymarin (100 mg/kg) + CCl₄ (10 ml/kg); IV: HAAD (50 mg/kg) + CCl₄ (10 ml/kg); V: HEAD (100 mg/kg) + CCl₄ (10 ml/kg); VI: HAAD (200 mg/kg) + CCl₄ (10 ml/kg). Values are mean ± S.E.M. (n=6). ^#p<0.05 vs. group (I); ^*p<0.05 vs. CCl₄ group (II

Figure 7.

Effect of HAAD and silymarin on total MDA levels in CCl₄-induced hepatotoxicity. Groups – I: normal saline; II: CCl₄ (CCl₄ group); III: Silymarin (100 mg/kg) + CCl₄ (10 ml/kg); IV: HEAD (50 mg/kg) + CCl₄ (10 ml/kg); V: HAAD (100 mg/kg) + CCl₄ (10 ml/kg); VI: HAAD (200 mg/kg) + CCl₄ (10 ml/kg). Values are mean ± S.E.M. (n=6). ^##p<0.01 vs. group (I); ^**p<0.01 and ^*p<0.05 vs. CCl₄ group (II

Figure 8

Effect of HAAD and silymarin on SOD, CAT, GST, and GSH activity in CCl₄-induced hepatotoxicity. Groups – I: normal saline; II: CCl₄ (CCl₄ group); III: Silymarin (100 mg/kg) + CCl₄ (10 ml/kg); IV: HAAD (50 mg/kg) + CCl₄ (10 ml/kg); V: HAAD (100 mg/kg) + CCl₄ (10 ml/kg); VI: HAAD (200 mg/kg) + CCl₄ (10 ml/kg). Values are mean ± S.E.M. (n=6). ^##p<0.01 and ^###p<0.001 vs. group (I); ^*p<0.05, ^**p<0.01, and ^***p<0.001 vs. CCl₄ group (II

Figure 9

Effect of HAAD on the histopathological morphology of rats liver as assessed by hematoxylin and eosin (H & E) staining (magnification X400). (A) Normal control, (B) CCl₄ group, (C) Silymarin 100 mg/kg b.w. + CCl₄, (D) HEAD 100 mg/kg b.w. + CCl₄, and (E) HAAD 200 mg/kg b.w. + CCl₄