Morphometric Evaluation of the Seminiferous Tubules and the Antioxidant Protective Effects of Gallic Acid and Quercetin in the Testis and Liver of Butyl Phthalate Treated Rats

Abstract

The antioxidant protective effects of gallic acid (GAL) and quercetin (QUE) against oxidative stress induced by di-butyl phthalate (DnBP) in the liver and testis of rats were evaluated in this study. Adult albino Wistar rats (180-225 g) were treated with QUE or GAL (50 mg/kg) alone or in combination with DnBP (1 mL/kg) for 15 days. After treatment, tissue samples were taken for determination of glutathione and malondialdehyde levels, and superoxide dismutase and catalase activities. Serial sections of the testis and liver were stained with haematoxylin and eosin for microscopy and seminiferous tubular morphometry. As expected, DnBP induced oxidative stress was evident by increased malondialdehyde level in both organs. Co-treatment with GAL or QUE reversed the malondialdehyde by 45.42, 37.44 and 37.57%, 23.32% and catalase by 52.21, 70.15 and 85%, 38.14% in the testis and liver respectively whereas superoxide dismutase activity and glutathione level were differently modulated parallel to histopathological improvement in both tissues. The seminiferous tubular diameter, epithelial height, epithelial germ cell count and tubular length were significantly decreased by 11.09, 51.91, 40.65 and 11.10% respectively versus control values after DnBP treatments and were attenuated on co-treatment with GAL or QUE. Co-treatment with GAL afforded better protective effects in both tissues but QUE treatment alone appeared more effective than GAL on the investigated morphometric data. It seems likely that GAL or QUE prevented the tissue damage but the antioxidant profiles of the liver and testis are different in response to the oxidative stress.

Keywords: Antioxidant; Di-n-butyl phthalate; Gallic acid; Morphometry; Oxidative stress; Quercetin.

Fig. 1

Effects of quercetin (QUE) and gallic acid (GAL) on the level of glutathione (GSH) in the liver (a) and testis (b) of rats after 15-day of oral treatment with di-n-butyl phthalate (DnBP). Values are mean ± SD; n = 5, *p < 0.05 versus control; **p < 0.05 versus DnBP

Fig. 2

Effects of quercetin (QUE) and gallic acid (GAL) on the level of malondialdehyde (MDA) in the liver (a) and testis (b) of rats after 15-day of oral treatment with di-n-butyl phthalate (DnBP). Values are mean ± SD; n = 5, *p < 0.05 versus control; **p < 0.05 versus DnBP

Fig. 3

Effects of quercetin (QUE) and gallic acid (GAL) on the activity of superoxide dismutase (SOD) in the liver (a) and testis (b) of rats after 15-day of oral treatment with di-n-butyl phthalate (DnBP). Values are mean ± SD; n = 5, *p < 0.05 versus control; **p < 0.05 versus DnBP

Fig. 4

Effects of quercetin (QUE) and gallic acid (GAL) on the activity of catalase (CAT) in the liver (a) and testis (b) of rats after 15-day of oral treatment with di-n-butyl phthalate (DnBP). Values are mean ± SD; n = 5, *p < 0.05 versus control; **p < 0.05 versus DnBP

Fig. 5

Cross sections of the testes of treated and control animals at the end of study. Control (a) Testis of control animals showed no visible lesions; Testis of Di-n-butyl phthalate treated animals (b) showed depleted epithelial height (arrow head) and azoospermia in the tubular lumen (long arrow), cytoplasmic vacuoles (short arrow). Also the lumen appears wider (L). Testis of gallic acid treated animals (c) appears similar to control and showed normal seminiferous tubules (arrow head); Testis of gallic acid + di-n-butyl phthalate treated animals (d) showed normal seminiferous tubules (arrow head); seminiferous tubules of quercetin treated animals (e) are normal with no visible lesion seen (arrow head); Testis of quercetin + treated animals (c) appears similar to control with no visible lesion and showed normal seminiferous tubules with most of the germ cells well represented (arrow head). Hematoxylin and eosin, mag × 400

Fig. 6

Cross sections of the liver of animals at the end of study. Control (a) no visible lesion seen; di-n-butyl phthalate treated animals (b) severe periportal cellular infiltration by mononuclear cells (long arrow); degenerated hepatocytes (starred); Gallic acid treated rats (c): no visible lesion seen; Gallic acid + di-n-butyl phthalate treated animals (d): mild and diffuse cellular infiltration by mononuclear cells (long arrow); Quercetin treated rats (e): no visible lesions seen; Quercetin + di-n-butyl phthalate treated rats (f): mild cellular infiltration by mononuclear cells (long arrow). Hematoxylin and eosin, mag × 400

Fig. 7

Effects of quercetin (QUE) and gallic acid (GAL) on the plasma indices of liver functions of rats after 14-day of oral treatment with di-n-butyl phthalate (DnBP). Values are mean ± SD; n = 5, *p < 0.05 versus control; **p < 0.05 versus DnBP

Fig. 8

Molecular structures of tested phytochemicals a Gallic acid: (3,4,5-trihydroxybenzoic acid), b Quercetin: 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxychromen-4-one