Exploring the phytochemicals, antioxidant properties, and hepatoprotective potential of Moricandia sinaica leaves against paracetamol-induced toxicity: Biological evaluations and in Silico insights

Abstract

Thirteen components were identified in the methanol extract of Moricandia sinaica leaves (MSLE) through analysis utilizing HPLC-ESI-MS/MS., including flavonoids, anthocyanins, phenolic acids, and fatty acids. The methanol extract of M. sinaica leaves contained total phenolics and flavonoids (59.37 ± 2.19 mg GAE/g and 38.94 ± 2.72 mg QE/g), respectively. Furthermore, it revealed in vitro antioxidant properties as determined by the DPPH and FRAP assays, with respective IC50 values of 10.22 ± 0.64 and 20.89 ± 1.25 μg/mL. The extract exhibited a notable hepatoprotective effect in rats who experienced paracetamol-induced hepatotoxicity. When a dose of 250 mg/kg was given, there was a 52% reduction in alanine transaminase and a 30% reduction in aspartate transaminase compared to the group with the disease. Furthermore, it demonstrated a 3.4-fold, 2.2-fold, and 2.6-fold increase in superoxide dismutase, non-protein sulfhydryl, and glutathione peroxidase, respectively. In addition, it demonstrated a 68% decrease in lipid peroxide levels compared to the group with paracetamol-induced condition. The verification was conducted using a histological study, which identified improved liver histology with a small number of distended hepatocytes. Moreover, in silico studies focused on the enzymes NADPH oxidase, butyrylcholinesterase, and tyrosinase as the targets for the major compounds. In conclusion, MSLE showed promising hepatoprotective and antioxidant activities due to its richness in antioxidant metabolites.

Fig 1. The chemical structures of major compounds of the M. sinaica leaves extract (MSLE) identified by HPLC-MS/MS.

Fig 2. The effect of M. sinaica leaves extract (MSLE) identified on serum liver function markers in PAR-induced liver toxicity in rats.

(a) Alanine aminotransferase, (ALT) (b) Aspartate aminotransferase (AST). Data are presented as mean ± SD (n = 6); * p < 0.05 vs. control rats; # p < 0.05 vs. PAR-treated rats using Tukey’s post hoc test.

Fig 3. The effect of M. sinaica leaves extract (MSLE) identified on oxidative stress markers in PAR-induced liver toxicity in rats.

(a) TBARs, (b) GSH, (c) SOD, and (d) GPx. Data are presented as mean ± SD (n = 6); * p < 0.05 vs. control rats; # p < 0.05 vs. PAR-treated rats using Tukey’s post hoc test.

Fig 4. Hematoxylin & eosin-stained sections from livers of all study groups, magnification X400.

(A & B) Group I (control) & II (MSLE) respectively. (C) Group III (PAR). (D) Group IV (Treatment with MSLE). (E) Group V (Treatment with NAC). C.V: Central vein, H: Hepatocytes, S: Blood Sinusoids, Black arrows: Ballooning of hepatocytes, Red arrows: Apoptotic hepatocytes, Blue arrows: Lipid droplets, Yellow arrows: Inflammatory cells. Circles: Lobular necrosis.

Fig 5. 2D binding modes of (A) Syringetin-3-O-glucoside and (B) Kaempferol-3,7-O-bis-α-L-rhamnoside to the active binding sites of NADPH enzyme.

Fig 6. 2D binding modes of (A) Cyanidin rutinoside and (B) Isorhamnetin to the active binding sites of BChE enzyme.

Fig 7. 2D binding modes of (A) Linarin and (B) 5,7-Dihydroxy-2’-methoxy-3’,4’-(methylenedioxy) isoflavone to the active binding sites of Tyrosinase enzyme.