Research On The Effect Of H₂O₂ On Apoptosis Of Stertoli Cells And The Machanism

Sertoli cells play a very important role in spermatogenesis and the quantity is the key of amount of spermatogenesis in adult animals. All poison aimed tesis play role at sertoli cells primary. Many study make clear that lots of poison aimed tesis have a close relationship with oxidative stress. H₂O₂ simulate oxidative stress in cltural sertoli cells of young swine. The effect of H₂O₂ on apoptosis of stertoli cells and the mechanism was studied. Accroding to the study, regulatory mechanism of microenvironment in spermatogenesis and the theory of spermatogenesis will be more clear. The study can also give us new thought about male sterility due to damage sertoli cells caused by oxidative stress.The effect of H₂O₂ on sertoli cells viability at different dose is studied to make a apoptosis model after oxidative stress. The results indicate that with the increasing dose of H₂O₂, sertoli cells atrophy, the amount of tentacles decrease, refraction become weak, lipid droplet decrease and the viability of sertoli cells descent. To make it clear that these change have a close relationship with H₂O₂, the effect of H₂O₂ on ROS in sertoli cells is detected. The results indicate that the level of ROS is upgrade with the increasing dose of H₂O₂. Superoxide anion increased at 0-800umol/L H₂O₂, which make it more clear that H₂O₂ have great effect on ROS.The study indicate that viability of sertoli cells descend gradually with the time in 0-24h. Cells viability at 48h have a no-marked change compared to that at 24h. So we take 600μmol/L H₂O₂ at 24h as suitable dose and time to get a apoptosis model.Sertoli cells have much morphological changes under light microscope, such as irregular intercellular space, maldistribution density distribution, weak refraction, degeneration, decreasing tentacles and morphological changes under electron microscope such as corrugativus cell membrane, oncotic mitochondrion, apoptotic body, which can be regarded as character of apoptosis. The study also detects the change of viability of Caspase3. The results make it clear that Caspase3 increased at high dose of H₂O₂, which indicate that Caspase3 dependet pathway play a important role in sertoli cells apoptosis due to H₂O₂ treatment.The effect of H₂O₂ on secretary function of sertoli cells is also detected. H₂O₂ make the level of transferrin significantly decrease, which mean that when H₂O₂ induced apoptosis of sertoli cells, it degrade the secretary function of sertoli cells. To study the mechanism of apoptosis of sertoli cells treated by H₂O₂, the effect of H₂O₂ on antioxidant ability of sertoli cells. The results indicate that H₂O₂ make the antioxidant material such as SOD,GSH,CAT significantly decreased. So the antioxidant ability due to H₂O₂ treatment may play a important role in apoptosis of sertoli cells.Many studies indicate that apoptosis induced by ROS may have relationship with MAPK. The study detected the phosphorylation of ERK1/2 and P38 of sertoli cells after treated by H₂O₂. We found that ERK1/2 and P38 are activated at different degree, which indicate that both ERK1/2 and P38 participate the apoptosis of sertoli cellsIn conclusion, we can get that(1) ROS in sertoli cells significantly increase after treated by H₂O₂, antioxidant ability of sertoli cells degrade, meanwhile the viability of sertoli cells significantly decrease and the ratio of apoptosis significantly increase.(2) Morphological changes happened in sertoli cells after treated by H₂O₂ such as irregular intercellular space, maldistribution density distribution, weak refraction, degeneration, decreasing tentacles and corrugativus cell membrane, oncotic mitochondrion, apoptotic body, crescent-shaped nucleus. Meanwhile, the secretary function of sertoli cells degrade.(3) H₂O₂ induced apoptosis of sertoli cells by caspase3 dependent pathway. Both ERK1/2 and P38 participate the apoptosis of sertoli cells.