Research On The Effect Of ROS On Cytoskeleton In Stertoli Cells

Sertoli Cell(SC)is critical to the spermatogenesis,and it is the first aim when the toxicant is invading the testis.The blood-testis-barrier,sperm releasing and materies transport are all related to the Sertoli Cell.The influence of the cytoskeleton reaches into almost every aspect of eukaryotic cell function.It is a little surprise therefore that links between the regulation of the cytoskeleton and oxidative stress have been found in a variety of eukaryotic systems.To illuminate the cytoskeleton how to respond to environmental signals,we use CytB and H₂O₂ to treat the Sertoli to simulate oxidative stress in vivo.To set up the property experiment modle.The Sertoli Cell is treated with H₂O₂.First of all,definiting the median lethal dose(LD50)at 600μmol/L through acute experiment.Accordingly,we detect the level of ROS.Compare to the control,the level is increased(p＜0.05).Meanwile,we dye the treated cell with DHE to detect the distribution of superoxide anion,photodensity is increased.ROS also change the ultrastructure: chondriosome swelling;chromatinclumping;endochylemacondensed;nucleus split and especially vacuolization.All of these changs are following with the variety of F-actin which stained with fluorescent antibody.Alao Sertoli Cells are treated with CytB to simulate oxidative stress in vivo as follows:0.1,0.5,1, 5μmol/L.The results are:the level of ROS is increased when the concentration of CytB is increased(p＜0.05),the tendency of photodensity change is equal to ROS.When the cells are treated with 0.1,0.5μmol/L CytB,F-actin is accumulated but the cell shape is intact.When reach to 5μmol/L,F-actin is broken,even chipping.To study the relationship between CytB and H₂O₂.we design as follow:600μmol/L H₂O₂,0.5μmol/L CytB,600μmol/L H₂O₂+0.5μmol/L CytB,600μmol/L H₂O₂+0.5μmol/L CytB+VE.It shows that they are synergistic effect,when adding VE to it,the effect can be weaken. To study the effect of ROS on the oxidative stress and antioxygen ability in Sertoli cells.We found CAT,GSH-Px,T-SOD and TAOC descended while the cell is treated differently.But when VE is added, antioxygen ability is increased(p＜0.05).To research the effect of ROS on the oxidative stress and antioxygen ability in Sertoli cells.We study the antioxygen ability and the activity of the MAPK.We found CAT,GSH-Px,T-SOD and TAOC descended while the cell is treated differently.But when VE is added,antioxygen ability is increased(p＜0.05).The result also shows that the level of ERK1/2 and P38 are advanced after treatment with CytB and/or H₂O₂.On the whold,we get these conclusion:(1)CytB and/or H₂O₂ invoke the increase of the level of ROS in Sertoli cell and decreas the viability of the Sertoli cell;(2)CytB arid/or H₂O₂ change the distribution of the F-actin;(3)CytB and/or H₂O₂ cause oxidative stress and cut down the antioxygen ability of cells. Besides,the activity of ERK1/2 and P38 are actived when adding VE,the effect is decreased.