The effect of salubrinal on er stress pathway in heat-stressed spermatogonicells al in vitro

Aim: It was aimed to investigate the effect of SAL on endoplasmic reticulum (ER) stress on experimental in vitro heat stress model (HSM) of spermatogenic cells. Materials and Methods: In the study Mouse spermatogonium (GC1) and spermatocyte (GC2) cell lines were used. IC50 dose of SAL were calculated via MTT assay. Each cell lines were separated into 4 different groups which are; control (GC1-C, GC2-C), SAL treated (GC1-SAL, GC2-SAL), HSM experimental model (GC1-HSM, GC2-HSM) and SAL treated HSM model (GC1-HSMSAL, GC2-HSMSAL). Control cells were incubated under standard culture conditions. IHM model group cells were incubated under 43oC for 60 minutes. In the SAL group, cells were incubated with 20 µM SAL containing culture medium for 24 hours. After the experiments, all groups were stained with immünoflorescence probes of p-PERK, ATF6, GRP78, p-IRE1α, p-eIF2α, HSP70 antibodies. Moreover, mRNA levels of GRP78, PERK, eIF2α were evaluated with qRT-PCR. Results: It was observed that HSM model has cytotoxicity. In the experimental model of HSM cells, all the markers showed elevated levels of immünoreactivity, that they were agreed upon to have ER stress. With the treatment of SAL, decreased levels of ER stress detected. GRP78, PERK and eIF2α mRNA levels were upregulated in the HSM model group. Even though there was a downregulation following SAL treatment, the difference was not statistically significant. Conclusions: In the view of such findings, it was thought that heat stress triggered ER stress on spermatogenic cells, and SAL has effect on these markers. Studies of ER-related stress mechanisms on spermatogenic cells will be important for the therapeutical strategies with the forthcoming advanced molecular analyses.