Shaygannia Erfaneh, Nasr-Esfahani Mohammad H, Sotoodehnejadnematalahi Fattah, Parivar Kazem
Department of Biology, School of Basic Sciences, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Department of Animal Biotechnology, Reproductive Biomedicine Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran.
Basic Clin Androl. 2021 Apr 1;31(1):10. doi: 10.1186/s12610-021-00125-9.
Ferroptosis 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.
The 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.
Despite 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.
铁死亡是一种与凋亡不同的铁依赖性细胞死亡。基于精索静脉曲张（VCL）大鼠体内过量的铁和活性氧，我们推测铁死亡可能参与了VCL的发生。此外，由于α-硫辛酸（ALA）具有抗氧化和抗铁死亡活性，我们在本研究中评估了精索静脉曲张模型中铁死亡的状态以及ALA的保护作用。为此，将70只雄性Wistar大鼠分为7组：对照组、假手术组和精索静脉曲张组，最初在手术后2个月处死以验证精索静脉曲张的诱导情况。在精索静脉曲张诱导后4个月处死第二批相同的3组大鼠，以评估补充ALA的效果。测量的参数包括染色质完整性（苯胺蓝和吖啶橙染色）、脂质过氧化（BODIPY染色）、睾丸形态测量和铁含量。此外，在术后2个月和4个月评估氧化还原（谷胱甘肽和烟酰胺腺嘌呤二核苷酸磷酸）和铁死亡（核因子E2相关因子2、溶质载体家族7成员11、P53和磷酸化c-Jun氨基末端激酶）标志物。
精索参数的改变证实了精索静脉曲张的诱导。精索静脉曲张期间睾丸中铁大量蓄积，ALA治疗后显著减少。铁死亡分子标志物在mRNA和蛋白质水平上没有显著改变。补充ALA没有改变烟酰胺腺嘌呤二核苷酸磷酸值，但提高了谷胱甘肽水平。
尽管在手术诱导精索静脉曲张后2个月和4个月睾丸中铁蓄积增加，但分子证据并未证明铁死亡的参与。这可能是由于精索静脉曲张的镶嵌性性质，影响了一些生精小管而不是其他生精小管，从而可能掩盖了分子标志物的变化。同时，我们的研究证实ALA刺激核因子E2相关因子2通路。
