Is ferroptosis involved in ROS-induced testicular lesions in a varicocele rat model?

Erfaneh Shaygannia,Kazem Parivar,Mohammad H Nasr-Esfahani,Fattah Sotoodehnejadnematalahi

doi:10.1186/s12610-021-00125-9

Abstract

BackgroundFerroptosis is an iron-dependent cell death that is distinct from apoptosis. Based on excessive amounts of iron and reactive oxygen species in varicocele (VCL) rats, we hypothesize that ferroptosis might be involved in VCL. In addition, since alpha-lipoic acid (ALA) was shown to have both antioxidant and anti-ferroptotic activity we assessed in the present work the status of ferroptosis in our varicocele model and the protective effect of ALA. To this end, 70 male Wistar rats were divided into 7 groups: control, sham and varicocele groups which were initially sacrificed 2 months after the operation to verify the induction of varicocele. A second batch of the same 3 groups were sacrificed 4 months after varicocele induction to evaluate the effect of ALA supplementation. The parameters measured were chromatin integrity (aniline blue and acridine orange staining), lipid peroxidation (BODIPY staining), testicular morphometry and iron content. In addition, redox (GSH and NADPH) and ferroptosis (Nrf2, Slc7a11, P53 and p-Jnk) markers were evaluated at 2 and 4 months post-operation.ResultThe alteration of the spermatic parameters made it possible to verify the induction of the varicocele. Iron accumulated well in the testicles during varicocele and decreased significantly following ALA treatment. Ferroptotic molecular markers at the mRNA and protein levels were not significantly altered. ALA supplementation did not alter NADPH values, but increased GSH levels.ConclusionDespite the increased accumulation of iron in the testes 2 and 4 months after surgical induction of varicocele, molecular evidence did not demonstrate the involvement of ferroptosis. This could be explained by the mosaic nature of the varicocele affecting some seminiferous tubules and not others which could mask variations in molecular markers. In parallel, our study confirms that ALA stimulates the NRF2 pathway.

Highlights

Programmed cell death or apoptosis plays a fundamental role in development, tissue homeostasis and various diseases
The same observations apply to animals at 4 months of age (V.I-alpha-lipoic acid (ALA)−; Fig. 2b) with significantly lower sperm concentration, higher abnormal morphologies and lower motility compared to the control group and/or the sham animals
It is interesting to note that when ALA was administered to the animals (V.I-ALA+), a significant improvement of these 3 parameters was observed with a sperm concentration going up, abnormal morphology going down, and sperm motility going up

Summary

Introduction

Programmed cell death or apoptosis plays a fundamental role in development, tissue homeostasis and various diseases. Another process of programmed cell death, genetically and biochemically distinct from apoptosis, has been described [1, 2] This new cell death process called ferroptosis is iron-dependent and is further characterized by an accumulation of iron-dependent lipid peroxides [1, 2]. Since alpha-lipoic acid (ALA) was shown to have both antioxidant and anti-ferroptotic activity we assessed in the present work the status of ferroptosis in our varicocele model and the protective effect of ALA. To this end, 70 male Wistar rats were divided into 7 groups: control, sham and varicocele groups which were initially sacrificed 2 months after the operation to verify the induction of varicocele. Redox (GSH and NADPH) and ferroptosis (Nrf, Slc7a11, P53 and p-Jnk) markers were evaluated at 2 and 4 months post-operation

Objectives

Results

Discussion

Conclusion