Research On The Protective Effect Of A-lipoic Acid To The Oxidative Stressed Sertoli Cells Of Rat Testis

Objective:To explore the protective effect of a-lipoic acid to the oxidative stressed sertoli cells of rat testis,and investigate expression of skelemin and their relevant molecular mechanisms in sertoli cells of rat testis oxidative stress procedure and a-lipoic acid protection.Methods:Primary cultured sertoli cells from rat testis and identified by the morphological changes, Sertoli cells were divided into7groups.They were seven oxidative groups injured by H2O2with different concentration (0,50,100,200,400,800,1600μmol/L,respectively). Sertoli cells viability was assessed by CCK-8.Moreover, intracellular reactive oxygen species(ROS) levels were measured by the fluorescent probe DCFH-DA. Primary rat sertoli cells were randomly divided into four groups,control,LA(200μmol/L),H2O2(400μmol/L)and H2O2(400μmol/L)plus LA(200μmol/L) pretreatment groups.Cell viability was assessed by CCK-8.Intracellular reactive oxygen species(ROS) levels were measured by the fluorescent probe DCFH-DA. The activities of antioxidant enzymes were measured by enzyme activity detection kit. The expressions of vimentin and a-tubulin were analyzed by immunohistochemical method and Western blot.Results:Sertoli cells of rat testis were integrated into a single layer after3-4days of culture.The H2O2groups were significantly different from the normal.The levels of CCK-8(OD) of cells in H2O2groups were significantly lower than those in the normal control group (P<0.05),and increased intracellular ROS,sertoli cells were interfered with400μmol/L H2O2.Decreased cell viability, increased intracellular ROS,reduced activities of antioxidant enzymes and the expressions of vimentin and a-tubulin were observed after the stimulation of H2O2(P <0.05).Pretreatment with LA improved cell viability and activities of antioxidant enzymes(as T-AOC,SOD,CAT and GR), decreased intracellular ROS levels.LA boosted the expressions of vimentin and a-tubulin induced by H2O2(P<0.05).Conclusion:H2O2induces the expressions of vimentin and a-tubulin and cell survival rate in sertoli cells. The protective role of LA in this induced damage through eliminating intracellular ROS and improved activities of antioxidant enzymes and the expressions of vimentin and a-tubulin.