Non-apoptotic cell death such as pyroptosis, autophagy, necroptosis and ferroptosis acts as partners to induce testicular cell death after scrotal hyperthermia in mice.

Cell death is a biologically uncontrollable and regulated process associated with human diseases which usually occur in response to oxidative stress that activates signalling pathways in multiple forms and can therefore contribute to human diseases. Thus, the current study aims to evaluate the signalling pathway involved in cell death after testicular hyperthermia. For this purpose, 32 mice were equally divided into four groups; I: Control; II, III and IV, Scrotal hyperthermia in which the testes are exposed to water at 43°C for 20 min every other day, respectively, 15, 10 and 5 times. Then, animals were euthanized and testicular tissue samples were isolated to evaluate protein expression as well as germ cell gene marker expression by Western blot and real-time PCR tests. Our data showed that the protein expression of Caspase-1, Beclin1, Atg7, Mlkl and Acsl4 together with the expression of Caspase-1, Beclin1, Atg7, Mlkl and Acsl4 genes was significantly up-regulated in scrotal hyperthermia-induced mice. In conclusion, the present study showed that heat stress disrupts spermatogenesis by activating several non-apoptotic signalling pathways in testicular tissue.

Ameliorating effects of curcumin-loaded superparamagnetic iron oxide nanoparticles (SPIONs) on the mouse testis exposed to the transient hyperthermia: A molecular and stereological study.

BACKGROUND: Testicular hyperthermia can have negative effects on male fertility. Despite reported therapeutic benefits of curcumin, several factors often limit its application such as low water solubility and instable structure. Curcumin-loaded superparamagnetic iron oxide nanoparticles (SPIONs) were designed to solve its limitation of use. In the present study, we evaluated the effect of curcumin-loaded SPIONs on transient testicular hyperthermia in mouse. MATERIALS AND METHOD: A total of 18 adult male NMRI mice were divided into three groups (n = 6): I. Controls (Cont), II. Scrotal hyperthermia (Hyp), III. Scrotal hyperthermia + curcumin-loaded iron particles (240 µL) (Hyp + Cur). After seventy days, the animals were sacrificed and used for further molecular and stereological evaluations. RESULTS: Sperm count, motility and viability significantly decreased in group hyp as compared to cont group. Furthermore, Sperm DNA fragmentation and cell apoptosis in testes increased remarkably in group hyp, compared with group cont. Stereological study showed a reduction in number of spermatogenic and Leydig cells, as well as reduced weight and volume of testes in hyp group. Degenerative appearance of testes exposed to hyperthermia was also observed. In addition, higher mRNA expression of inflammatory cytokines (IL1-α, IL6, and TNF-α) was detected in group hyp compared to cont group. However, curcumin-loaded SPIONs alleviated all of the pathologic changes in the Hyp + Cur group compared to the hyp group. CONCLUSION: Here, we used nanoparticle form of curcumin in testicular hyperthermia model and showed its ameliorating effects on testes damages caused by heat stress, which can be an appropriate method to overcome the problems that limit curcumin application in cases with increased intra testicular temperature.