Efeito protetor do extrato hidroalcoólico de agaricus blazei murill em testículos de camundongos expostos ao quimioterápico 5-fluorouracil

Abstract

The 5-Fluorouracil (5-FU) is a chemotherapeutic agent capable of inducing infertility, compromising sperm viability and testicular tissue (TT) morphology. In this context, in the search for bioactive compounds with protective effects that may mitigate these adverse outcomes, the hydroalcoholic extract (HEAb) of the fungus Agaricus blazei Murill emerges as a potential candidate due to its anti-inflammatory and antioxidant effects in other tissues. However, no studies have investigated this compound in the male reproductive system. Thus, the aim of the present study was to evaluate the protective effect of HEAb against 5-FU toxicity on sperm viability and TT morphology in Swiss mice. For this purpose, 18 mice were subjected to three treatments: (1) Control (NaCl 0.9%); (2) 5-FU 450 mg/kg/single dose; (3) 5-FU 450 mg/kg/single dose + HEAb 25 mg/kg/3 days. After three days, the animals were euthanized, and the TT were fixed for classical histology (HE), sperm viability, cell proliferation (AgNOR), quantification of MDA and GSH to assess antioxidant activity, immunostaining for MMP-2 and MMP-9, and DNA fragmentation by TUNEL assay. Animals treated with 5-FU showed morphological alterations, reduced sperm viability and proliferative capacity of spermatogonia and Sertoli cells, and increased oxidative stress compared to the control group (p < 0.0001). However, HEAb administration preserved testicular morphology and sperm viability similar to the control, increased cell proliferation compared to the 5-FU group (p < 0.001), and reduced oxidative stress compared to the other treatments (p < 0.001). Regarding immunostaining, the 5-FU group showed a significant increase in MMP-2 and MMP-9 expression compared to the control group (p < 0.001), while the group treated with co-administration of HEAb also showed increased MMP-2 expression but reduced MMP-9 expression (p < 0.001). DNA fragmentation analysis by TUNEL revealed a significant increase in apoptosis rates in germ cells of the 5-FU group compared to the control (p < 0.001), whereas the 5-FU+HEAb group showed a significant reduction in positive TUNEL labeling compared to the 5-FU group (p < 0.05). No significant differences were observed between the 5-FU+HEAb group and the control group (p > 0.05). These findings indicate that the hydroalcoholic extract of A. blazei Murill was able to mitigate the toxic effects of 5-FU on morphology, sperm viability, oxidative stress, cell proliferation, matrix remodeling, and DNA fragmentation.