Nephroprotective Effects of Fraxinus Hookeri Wenz. Against Renal Toxicity and DNA Oxidative Damages Induced by CCl4 in Rats

Abstract

Recognizing the therapeutic value of the Genus Fraxinus worldwide, this study evaluates the antioxidant potential of Fraxinus hookeri Wenz. (F. hookeri) against CCl₄-induced nephrotoxicity in rats. Forty-eight rats are randomly allocated into eight groups (six rats each). Antioxidant enzymes, genotoxicity, urine and serum markers, and tissue histopathology are assessed to determine their nephroprotective effects. The Control group remains untreated, while the DMSO group receives vehicle olive oil intraperitoneally and DMSO orally (3 ml/kg). All other groups, except Control and DMSO, are given CCl₄ (3 ml/kg, i.p., in 30% olive oil) twice weekly for 4 weeks. The CCl₄ group receives only CCl₄. The Rutin group receives reference drug Rutin orally (50 mg/kg). MEFH100 and MEFH200 groups are given MEFH at 100 and 200 mg/kg, respectively, and NHFH100 and NHFH200 receive NHFH at the same doses. Rutin and F. hookeri treatment effectively (P < 0.05) restore urine and serum markers disrupted by CCl₄. CCl₄ reduced (p < 0.05) antioxidant enzymes (CAT, SOD, POD) and increased TBAR levels and DNA damage, which are reversed by cotreatment with F. hookeri and Rutin. Histopathological improvements (P < 0.05) are also observed with F. hookeri. The results indicate that F. hookeri enhances antioxidant defenses, supporting its potential against CCl₄-induced nephrotoxicity.

Keywords: Fraxinus hookeri; CCl4; genotoxicity; histopathology; nephroprotective; nephrotoxicity; renal.

Figure 1

Microscopic images of H&E‐stained kidney sections representing all research groups. A) Control group, B) DMSO + Olive oil, C) CCl₄, D) Rutin + CCl₄, and F. hookeri extracts Groups E) MEFH₁₀₀ + CCl₄, F) MEFH₂₀₀ + CCl₄, G) NHFH₁₀₀ + CCl₄, and H) NHFH₂₀₀ + CCl₄. The magnification for all other groups is 40×. Glomerulus (dark green arrow), Mesangium (orange arrow), Bowman's capsule (dark blue arrow), Bowman's space (light green arrow), Distal tubules (light blue arrow), Proximal tubules (yellow arrow), Thick wall vein (pink arrow).

Figure 2

Evaluation of the nephroprotective effects of F. hookeri different fractions on urine profile levels, including urobilinogen, creatinine, albumin, creatinine clearance, and protein in rats, with Mean ± SD shown for every group (n = 6). ++ indicate significance from the CCl₄ group at P < 0.05 and probability level is P < 0.01. ** indicate significance from the Control group at P < 0.05 and probability level is P < 0.01.

Figure 3

Evaluation of the nephroprotective effects of F. hookeri different fractions on urine profile levels, including urinary urea, specific gravity, RBC, PH, and WBC in rats, with Mean ± SD shown for every group (n = 6). ++ indicate significance from the CCl₄ group at P < 0.05 and probability level is P < 0.01. ** indicate significance from the Control group at P < 0.05 and probability level is P < 0.01.

Figure 4

In rats, the nephroprotective effects of F. hookeri different fractions on serum profile levels, including serum protein, globulin, serum urea, albumin, and serum nitrite, with Mean ± SD shown for every group (n = 6). ++ indicate significance from the CCl₄ group at P < 0.05 and probability level is P < 0.01. ** indicate significance from the Control group at P < 0.05 and probability level is P < 0.01.

Figure 5

Nephroprotective effects of F. hookeri different fractions on serum profile levels, including direct bilirubin, total bilirubin, creatinine, urobilinogen, and creatinine clearance in rats, with Mean ± SD shown for every group (n = 6). ++ indicate significance from the CCl₄ group at P < 0.05 and probability level is P < 0.01. ** indicate significance from the Control group at P < 0.05 and probability level is P < 0.01.

Figure 6

Evaluation of the nephroprotective effects of F. hookeri different fractions on the enzymatic antioxidant levels in rats, with Mean ± SD shown for every group (n = 6). ++ indicate significance from the CCl₄ group at P < 0.05 and probability level is P < 0.01. ** indicate significance from the Control group at P < 0.05 and probability level is P < 0.01.

Figure 7

Agarose gel displays the nephroprotective properties of different F. hookeri fractions and the DNA damage caused by CCl₄. From left to right lanes (L) low molecular weight marker, (i) Control, (ii) DMSO, (iii) CCl₄, (iV) Rutin, and F. hookeri extracts Groups: (V) MEFH₁₀₀ + CCl₄, (Vi) NHFH₁₀₀ + CCl₄, (Vii) MEFH₂₀₀ + CCl₄, and (Viii) NHFH₂₀₀ + CCl₄.

Figure 8

Evaluation of the nephroprotective effects of F. hookeri by comparing several renal DNA fragmentation percentage levels across various study groups of rats. Mean ± SD is displayed for each group (n = 6). ++ indicate significance from the CCl₄ group at P < 0.05 and probability level is P < 0.01. ** indicate significance from the Control group at P < 0.05 and probability level is P < 0.01.