GC-MS analysis and hepatoprotective activity of the n-hexane extract of Acrocarpus fraxinifolius leaves against paracetamol-induced hepatotoxicity in male albino rats

Abstract

Context: In Egypt, the burden of liver diseases is exceptionally high.

Objective: To investigate the components of the n-hexane extract of Acrocarpus fraxinifolius Arn. (Leguminosae) and its hepatoprotective activity against paracetamol (APAP)-induced hepatotoxicity in rats.

Material and methods: TRACE GC ultra gas chromatogaphic spectrometry was used for extract analysis. Thirty albino rats were divided into six groups (five rats in each). Group 1 was the healthy control; Groups 2 and 3 were healthy treated groups (250 and 500 mg/kg b.w. of the extract, respectively) for seven days. Group 4 was hepatotoxicity control (APAP intoxicated group). Groups 5 and 6 received APAP + extract 250 and APAP + extract 500, respectively.

Results: Chromatographic analysis revealed the presence of 36 components. Major compounds were α-tocopherol (18.23%), labda-8 (20)-13-dien-15-oic acid (13.15%), lupeol (11.93%), phytol (10.95%) and squalene (7.19%). In the acute oral toxicity study, the mortality rates and behavioural signs of toxicity were zero in all groups (doses from 0 to 5 g/kg b.w. of A. fraxinifolius). LD₅₀ was found to be greater than 5 g/kg of the extract. Only the high dose (500 mg/kg b.w.) of extract significantly alleviated the liver relative weight (4.01 ± 0.06) and biomarkers, as serum aspartate aminotransferase (62.87 ± 1.41), alanine aminotransferase (46.74 ± 1.45), alkaline phosphatase (65.96 ± 0.74), lipid profiles (180.39 ± 3.51), bilirubin profiles (2.30 ± 0.06) and hepatic lipid peroxidation (114.20 ± 2.06), and increased body weight (11.58 ± 0.20), serum protein profile (11.09 ± 0.46) and hepatic total antioxidant capacity (23.78 ± 0.66) in APAP-induced hepatotoxicity in rats.

Conclusion: Our study proves the antihepatotoxic/antioxidant efficacies of A. fraxinifolius hexane extract.

Keywords: Acetaminophen; gas chromatography mass spectrometry; ink cedar; liver dysfunction.

Figure 1.

GC-MS chromatogram of n-hexane extract of A. fraxinifolius leaves.

Figure 2.

Body weight gain (A) and liver relative weight (B) of control and intoxicated rats. Values are means, with their standard errors represented by vertical bars. A. fraxinifolius: Acrocarpus fraxinifolius; APAP: N-acetyl-p-aminophenol **p < 0.01, ***p < 0.001: compared with the healthy control group; †††p < 0.001: compared with the APAP-intoxicated group that received vehicle; (one-way ANOVA with Tukey’s multiple comparison test).

Figure 3.

Serum liver enzymes markers of control and intoxicated rats. Values are means, with their standard errors represented by vertical bars. A. fraxinifolius: Acrocarpus fraxinifolius; ALAT: alanine aminotransferase; ALP: alkaline phosphatase; APAP: N-acetyl-p-aminophenol; ASAT: aspartate aminotransferase *p < 0.05, **p < 0.01,***p < 0.001: compared with the healthy control group; †††p < 0.001: compared with the APAP-intoxicated group that received vehicle; (one-way ANOVA with Tukey’s multiple comparison test).

Figure 4.

Serum lipid profiles (A: total cholesterol and triglycerides, B: HDL-cholesterol and LDL-cholesterol, C: atherogenic index 1 and 2) of control and intoxicated rats. Values are means, with their standard errors represented by vertical bars. A. fraxinifolius: Acrocarpus fraxinifolius; APAP: N-acetyl-p-aminophenol Atherogenic index (1), total cholesterol: HDL-cholesterol ratio; atherogenic index (2), LDL-cholesterol: HDL-cholesterol ratio *p < 0.05, **p < 0.01, ***p < 0.001: compared with the healthy control group; †††p < 0.001: compared with the APAP-intoxicated group that received vehicle; (one-way ANOVA with Tukey’s multiple comparison test).

Figure 5.

Serum total protein, albumin and globulin levels (A) and A/G ratio (B) of control and intoxicated rats. Values are means, with their standard errors represented by vertical bars. A. fraxinifolius: Acrocarpus fraxinifolius; APAP: N-acetyl-p-aminophenol *p < 0.05, **p < 0.01, ***p < 0.001: compared with the healthy control group; †p < 0.05: compared with the APAP-intoxicated group that received vehicle; (one-way ANOVA with Tukey’s multiple comparison test).

Figure 6.

Serum total, direct and indirect bilirubin levels of control and intoxicated rats. Values are means, with their standard errors represented by vertical bars. A. fraxinifolius: Acrocarpus fraxinifolius; APAP: N-acetyl-p-aminophenol **p < 0.01, ***p < 0.001: compared with the healthy control group; †p < 0.05, †††p < 0.001: compared with the APAP-intoxicated group that received vehicle; (one-way ANOVA with Tukey’s multiple comparison test).

Figure 7.

Hepatic MDA concentration (A) and total antioxidant capacity (B) of control and intoxicated rats. Values are means, with their standard errors represented by vertical bars. A. fraxinifolius: Acrocarpus fraxinifolius; APAP: N-acetyl-p-aminophenol; MDA: malondialdehyde *p < 0.05, **p < 0.01, ***p < 0.001: compared with the healthy control group; †p < 0.05, †††p < 0.001: compared with the APAP-intoxicated group that received vehicle; (one-way ANOVA with Tukey’s multiple comparison test).