Efficacy of Quercetin-Sensitized Cisplatin against N-Nitroso-NMethylurea Induced Testicular Carcinogenesis in Wistar Rats

Abstract

Background: Testicular cancer is a public health problem. The goal of this study was to demonstrate the efficacy of quercetin treatment on N-nitroso-N-methyl-urea (MNU)-induced testicular carcinogenesis alone or in combination with cisplatin-treatment.

Methods: In total 70 adult male albino rats were categorized into six groups, control, quercetin-treatment (10 mg/kg body weight), cisplatin-treatment (2 mg/kg. body weight), cisplatin and quercetin-treatment, MNU-treatment, MNU plus quercetin-treatment and MNU plus quercetin and cisplatin-treatment. Treatment with quercetin and/or cisplatin was performed after 2 months of MNU induced testicular carcinogenesis. The studied groups were euthanized and sacrificed and their testes were examined for gene expression, biochemical, histological and immunohistochemically analysis, inflammation and apoptosis of germ cells.

Results: The fertility of the rats subjected to MNU carcinogenesis was impaired following cisplatin and/or quercetin-treatment. Cisplatin-treatment reduced the fertility rate and improved after quercetin-treatment. Quercetin-treatment decreased the sharp increase in RNA expression of BAX and MPO in both cisplatin-toxicated testes and after MNU carcinogenesis induction. In addition, the testicular levels of testosterone and SOD increased in parallel with depletion of MDA, IL-6, AFP and caspase-3 levels in MNU and/or cisplatin-treatment after -quercetin-treatment. The testicular structure of the cisplatin-treated group recovered their dividing germ and sperm differentiation after-quercetin-treatment. While, there was a great appearance of flourishing germ cell of MNU carcinogenesis post quercetin therapy, there was still a lack of sperm differentiation. Conclusion: Quercetin-treatment showed increased cisplatin activity and decreased testicular carcinogenesis due to anti-neoplastic and antioxidant activities.

Keywords: Cisplatin; Gene expression; MNU; Quercetin; Testicular cancer.

Figure 1

Chart Illustrating Experimental Design

Figure 2

Percentages of Body Weight Gain and Absolute Body Weight, Testis Weight and Relative Testis Weight of Male Rats Subjected to MNU Carcinogenesis and Treated with Cisplatin and/or Quercetin. Each result represents the mean + SD (n=7),* Significant at P<0.05. Abbreviations; C, control; Cis, cisplatin; Cis +Q, cisplatin and quercetin; MNU, methyl nitroso urea; MNU+Q; methyl nitroso urea and quercetin; MNU+Q+Cis; methyl nitroso urea plus quercetin plus cisplatin

Figure 3

Photomicrographs of Histological Cross Section of Rat Testis. A. Control showing active ST. B. Cisplatin intoxication showing reduction of germ cells and exfoliation of some within tubular lumina. C. Quercetin therapy of cisplatin intoxication showing regenerated spermatogenic cells. D&D1. MNU carcinogenesis showing atrophy and degeneration of germ cells. E. MNU carcinogenesis treated with quercetin showing regenerated spematogonial and spermatocyte cells. F. MNU carcinogenesis treated with cisplatin and quercetin showing regenerated germ cells but no presence of sperm. Abbreviations; ACT, active seminiferous tubules; AT, atrophied tubule; DSC, degenerated spermatocyte; EGC, exfoliation of germ cells; IST,imactive seminiferous tubule; SG, spermatogonia; Sg, spermatocyte

Figure 4

Photomicrographs of Formalin Fixed Testis Immunohistochemical Stain with PCNA (A-F), TNF-α (a1-F1), Caspase 3 (A2-F2) and PF3(A3-F3) Vertical. Note increased immunostaining of PCNA in control and improvement post quercetin-treatment of cisplatin and MNU carcinogenesis. TNF-α showing overexpression in cisplatin intoxication and MNU carcinogenesis and improved in quercetin-treated groups. Caspase intoxication showing increased immune reaction in cisplatin intoxication and MNU carcinogenesis and improved in quercetin-treated studied groups. P53 showing increased immune staining in cisplatin intoxication and MNU carcinogenesis and improved in quercetin-treated studied groups

Figure 5

Image Analysis of Male Testis Subjected to MNU Carcinogenesis and Treated Treated with Cisplatin and/or Quercetin. PCNA showing significant decreased image reaction in cisplatin and MNU carcinogens and/or quercetin-treatment. TNF-α expressing inflammation showing significant increase of image analysis in cisplatin and MNU carcinogens and/or quercetin-treatment. Caspase-3 expressing apoptosis showing significant increase image analysis cisplatin and MNU carcinogens and/or quercetin-treatment. P53 reflecting mutagenicity showing significant increase image analysis cisplatin and MNU carcinogens and/or quercetin-treatment. Stare mean significant at P ‹ 0.05

Figure 6

RNA Expression of Testis of Male Rat Subjected to MNU Carcinogenesis Testicular Tumor and Treated with Cisplatin and/or Quercetin Treatment. A. SOD gene expression. B. MPO gene expressio. C. BAX gene expression. D. Bcl2 gene expression. Each result represents the mean ± SD (n=4). Stare mean significant at P ‹ 0.05