The ferroptosis of sertoli cells inducing blood-testis barrier damage is produced by oxidative stress in cryptorchidism

Abstract

Oxidative Stress (OS) is the main cause of damage to the Blood-Testis Barrier (BTB) in cryptorchidism, which seriously endangers male reproductive health. It is well known that the OS induced ferroptosis is an important cause of dysfunction in the body. However, it is still unknown whether BTB damage in cryptorchidism leads to ferroptosis of Sertoli cells. We establishing the cryptorchidism model through surgery to avoid the complex effects of drugs on the model animals, combined with in vitro culture of the primary Sertoli cells for validation, and the methods of immunofluorescence staining, Western blotting and Prussian blue staining were used to study the oxidative stress in cryptorchidism. The effects of ferroptosis of Sertoli cells inducing BTB damage caused by OS in cryptorchidism were analyzed. We found that the inhibition of Nrf-2/keap-1/HO-1 pathway resulted in decreased expression levels of Glutathione Peroxidase 4 (GPX4), Ferroportin 1 (FPN1), and increased expression of Ferritin light chain (FTL) protein. Our research further confirms that inhibiting ferroptosis reduced BTB damage by reflecting a decrease expression of Zonula Occludens protein 1 (ZO-1), Occludin and Claudin-11 protein caused by OS. In addition, we found that the testosterone (T) secretion disorders and the supplementation of T can alleviate the damage of the BTB in cryptorchidism, and this effect is achieved through the Androgen Receptor (AR). In conclusion, our study found that the inhibition of Nrf-2/keap-1/HO-1 pathway in testis and the reduction of Tight junction proteins (TJs) ZO-1, Occludin and Claudin-11 protein expression levels in cryptorchidic mice, indicated that the cryptorchidism triggering a serious reproductive disorder, and one of the important reasons is the OS induced ferroptosis of Sertoli cells, which ultimately leads to the damage of the BTB. This findings may have important implications in the field of male reproductive disorders.