Oxidative stress mediated hepatotoxicity induced by ZNP and modulatory role of fruit extract on male Wistar rat

Abstract

Zinc oxide nanoparticles (ZNP) are being used in various fields viz cosmetics industry as UV protectants, in the food packaging industry due to their anti-bacterial properties, in agriculture as micronutrients, etc. Increased applications of ZNPs in our day to day life, leading to the contamination of the surrounding environment posing a direct or indirect health risk. Various reports suggest that fruits and vegetables are a rich source of phytochemicals having antioxidant properties which help in neutralizing ROS generated on metal toxicity of the body. The present study focuses to study the ameliorative effect of apple (Pyrus malus) extract (E) on ZNP induced toxicity. Therefore, animals were grouped, six in each, exposed to various doses of ZNP (50 and 250 mg/kg), ZNP (50 and 250 mg/kg)+E. The studied parameters was: food intake, water intake, antioxidants assay, zinc accumulation, and histological alterations and was compared to control. Investigation revealed that ZNP induces toxicity as revealed by the alteration in the studied parameter, whereas those exposed to ZNP along with Pyrus malus fruit extract try to reduce the toxicity induced by nanoparticles but at low doses only. This ameliorative effect of fruit extract might be due to the presence of antioxidants scavenging the free radicals generated by ZNPs suggesting that antioxidant-rich fruit may have a protective role and have the potential to reduce the nanoparticles mediated oxidative stress.

Keywords: Antioxidants; Fruit extract; Nanotoxicity; Oxidative stress; ZNP.

Fig. 1

Schematic representation of plan of work.

Fig. 2

a) UV–vis spectrum b) XRD c–d) SEM micrograph and SEM-EDX, e–f) TEM micrograph and Histogram of ZNP g) Zeta potential are the techniques used for the characterization of ZNP.

Fig. 3

Relative body weight change (%) of ZNP exposed rat treated with Pyrus malus fruit extract (E). *significant (p < 0.05). Where, control = received water, ZNP50= received 50 mg/kg ZNP, ZNP250= received 250 mg/kg ZNP, ZNP50E = received 50 mg/kg ZNP along with fruit extract (E), ZNP250E = received 250 mg/kg ZNP along with fruit extract (E), and E only = received fruit extract only.

Fig. 4

Effect of ZNP exposed male Wistar rat followed by Pyrus malus fruit extract (E) on average a) Zinc content b) liver coefficients c) LDH d) Water intake e) Food intake, and c) feces excretion after 28 days of treatment. Value expressed Mean ± SEM. *significant at (p < 0.05), **significant at (p < 0.01), ***significant at (p < 0.001). Where, control = received water, ZNP50= received 50 mg/kg ZNP, ZNP250= received 250 mg/kg ZNP, ZNP50E = received 50 mg/kg ZNP along with fruit extract (E), ZNP250E = received 250 mg/kg ZNP along with fruit extract (E), and E only = received fruit extract only.

Fig. 5

Effect of pharmacological intrusion on ZNP exposed rats treated with Pyrus malus fruit extract (E) reflected by Oxidative stress biomarkers a) a) NADPH Cyt P450 R b) NADH Cyt b5 R c) SOD activity d) CAT activity e) GPX activity f) GR activity. The result presented as Mean ± SEM. *Significant (p < 0.05) **Significant (p < 0.01), *** Significant (p < 0.001). Where, control = received DW, ZNP50=received 50 mg/kg ZNP, ZNP250= received 250 mg/kg ZNP, ZNP50E = received 50 mg/kg ZNP along with fruit extract (E), ZNP250E = received 250 mg/kg ZNP along with fruit extract (E), and E only = received fruit extract only.

Fig. 6

Histopathological changes of liver tissue stained with Hematoxylin and Eosin (at 400X) of Wistar rats treated with ZNP followed by Pyrus malus fruit extract (a) Control group showed normal architecture showing hepatocytes and sinusoids (b) ZNP(50 mg/kg) low dose results in slight distorted architecture, asterisks shows diallated sinusoids, Circles showed binucleated cell, (c) ZNP(250 mg/kg) high doses showed further dilation as compared to low doses increased, circle showed binucleated cell also arrows shows vacuolar degeneration (d) ZNP(50 mg/Kg)+Extract showed comparable less distorted cell architecture with to the groups received 50 mg/kg ZNP only and (e) ZNP(250 mg/Kg)+Extract circle shows binucleated cell, less dilated sinusoids as compared to those which received high dose of ZNP, vacuolation persist (f) Extract only showed normal architecture as in control.