Zinc oxide resveratrol nanoparticles ameliorate testicular dysfunction due to levofloxacin-induced oxidative stress in rats

Abstract

The present work is aimed to assess the protective influence of zinc oxide resveratrol nanoparticles against oxidative stress-associated testicular dysfunction. The number of 50 male albino rats were randomly separated into five groups (n = 10): Group I, control: rats gavage distilled water orally; Group II, Levofloxacin: rats that administered Levofloxacin (LFX) softened in distilled water at a dosage of 40 mg/kg^-1 BW orally every other day; Group III, Zn-RSV: rats administered with Zn-RSV (zinc oxide resveratrol in distilled water at a dose 20 mg/kg^-1 BW orally every other day; Group IV, (LFX + Zn-RSV): rats that were administered with Levofloxacin along with Zn-RSV nPs; Group V, Levofloxacin + Zn: rats were administered with Levofloxacin and Zno at a dose of 20 mg/kg^-1 BW orally every other day as mentioned before. This study lasted for 2 months. Sera were collected to assess luteinizing hormone (LH), follicle-stimulating hormone (FSH), and testosterone values. Testicular tissues were utilized to evaluate levels of superoxide dismutase (SOD), nitric oxide (NO), malondialdehyde (MDA), and catalase (CAT). Semen samples were utilized to measure their quality (motility, concentration, and vitality). Histopathological and immune histochemical techniques investigated the morphological changes in the testis. Rats treated with Levofloxacin showed significantly lower levels of serum LH, testosterone, FSH, testicular enzymatic NO, catalase, SOD, BAX, and BCL-2 immune reactivity and sperm quality but significantly greater testicular malondialdehyde and caspase-3 immuno-reactivity Compared to both control and zinc oxide resveratrol treatment. Zinc oxide resveratrol nanoparticles ameliorated the harmful side effects of Levofloxacin. Improvements were more pronounced in the co-treatment (LFX + Zn-RSV) Zinc oxide resveratrol group than in the co-treatment (LFX + Zno) Zinc oxide group. Zinc oxide resveratrol nanoparticles could be a possible solution for levofloxacin oxidative stress-induced fertility problems.

Figure 1

Transmission electron micrograph of RSV-Zno NPs. RSV-Zno nPs appeared as hexagonal nanostructures with crystallite size values consistent with XRD technique.

Figure 2

Characterization of the prepared Zno nPs with XRD. XRD pattern of the Zno nPs depicts the characteristic peaks of the hexagonal wurtzite P63mc crystal structure.

Figure 3

FT-IR Spectrum of the prepared Zno nPs. The FT-IR spectrum of the optimized Zno nPs revealed an obvious peak at 430 cm-1 assigned to Zn–O stretching.

Figure 4

photomicrograph, testes of rat (a) Control group showing normal histological structure of seminiferous tubules (ST). (b) Group treated with zinc oxide resveratrol nanoparticle showing normal histological structure of seminiferous tubules (ST). (c) Group treated with levofloxacin showing degeneration of seminiferous tubules (D) and congestion of tunica albuginea blood vessels (C). (d) Group treated with levofloxacin showing cysticaly dilated seminiferous tubules which devoid of spermatogonial cells (CD), spermatid giant cell in tubular lumen (short arrow) with severe congested tunica albuginea blood vessels (C). (e) Group treated with levofloxacin, note edema of interstitial tissue (E). (f) Group treated with levofloxacin showing congestion of interstitial blood vessel (C). (g) Group treated with levofloxacin, note thickening of tunica albuginea with congestion of blood vessels (C). (h) Group treated with levofloxacin and co-administered with zinc oxide resveratrol nanoparticle, note normal histological structure of seminiferous tubules (ST), mild congestion of interstitial blood vessels (C) and mild interstitial edema (E). (i) Testis of group treated with levofloxacin and co-administered with zinc oxide nanoparticle showing testicular degeneration (D). (j) Testis of group treated with levofloxacin and co-administered with zinc oxide nanoparticle, note interstitial edema (E) and congestion of tunica albuginea blood vessel (C). (H&E X200).

Figure 5

photomicrograph, epididymis of rat (a) Epididymis of control group showing normal histological structure of epididymal tubule with presence of sperms in the lumen (ET). (b) Epididymis of group treated with zinc oxide resveratrol nanoparticle showing normal histological structure of epididymal tubule (ET) and capsule (Ca). (c) Group treated with levofloxacin showing necrosis of tubular lining epithelium (N) with thickening of capsule (Th). (d) Group treated with levofloxacin showing edema of interstitial tissue with infiltration of mononuclear cells (E). (e) Group treated with levofloxacin and co-administered with zinc oxide resveratrol nanoparticle, note normal histological structure of epididymal tubule (ET) and capsule (Ca). (f) Group treated with levofloxacin and co-administered with zinc oxide nanoparticle, note necrosis of tubular lining epithelium (N) with congestion of capsular blood vessel (C). (g) Group treated with levofloxacin and co-administered with zinc oxide nanoparticle, note capsular edema and congestion (E). (h) Group treated with levofloxacin and co-administered with zinc oxide nanoparticle showing edema with mononuclear inflammatory cells infiltration in capsule (E) (H&E X200).

Figure 6

Immunostaining of BCL-2 and BAX in testes, (a) & (b) BCL-2 in control and zinc oxide resveratrol nanoparticle treated groups respectively showing no immune reactive cells (BCL-2 X200). (c) Levofloxacin treated groups showing weak positive expression (BCL-2 X200). (d) Group treated with levofloxacin and co-administered with zinc oxide resveratrol nanoparticle showing strong positive expression of BCL-2 (BCL-2 X200). (e) Group treated with levofloxacin and co-administered with zinc oxide nanoparticle showing weak positive expression of BCL-2 (BCL-2 X200). (f) & g) BAX in control and zinc oxide resveratrol nanoparticle treated groups respectively showing no immune reactive cells (BAX X200). (h) Levofloxacin treated groups showing strong positive expression of BAX (BAX X200). (i) Group treated with levofloxacin and co-administered with zinc oxide resveratrol nanoparticle showing weak positive immune expression of BAX (BAX X200). (j) Group treated with levofloxacin and co-administered with zinc oxide nanoparticle showing strong positive expression of BAX (BAX X200).

Figure 7

Immunostaining of BCL-2 and BAX in epididymis, (a) & (b) BCL-2 in control and zinc oxide resveratrol nanoparticle treated groups respectively showing no immune reactive cells (BCL-2 X200). (c) Levofloxacin treated groups showing weak positive expression (BCL-2 X200). (d) Group treated with levofloxacin and co-administered with zinc oxide resveratrol nanoparticle showing strong positive expression of BCL-2 (BCL-2 X200). (e) Group treated with levofloxacin and co-administered with zinc oxide nanoparticle showing weak positive expression of BCL-2 (BCL-2 X200). (f) & (g) BAX in control and zinc oxide resveratrol nanoparticle treated groups respectively showing no immune reactive cells (BAX X200). (h) Levofloxacin treated groups showing strong positive expression of BAX (BAX X200). (i) Group treated with levofloxacin and co-administered with zinc oxide resveratrol nanoparticle showing weak positive immune expression of BAX (BAX X200). (j) Group treated with levofloxacin and co-administered with zinc oxide nanoparticle showing strong positive expression of BAX (BAX X200).