Effects Of Vitexin On Streptozotocin-Induced Testicular Dysfunction And Its Underyling Mechanisms In Diabetic Mice

ObjectiveTo investigated the effects of vitexin on streptozotocin?STZ?-induced diabetic testicular dysfunction and its underyling mechanisms in male mice.Methods1.To investigate the protective effects of vitexin on diabetic testicular dysfunction in male mice.DM was induced by intraperitoneal injection of STZ 45 mg/kg for 5consecutive days.The animals were divided into six experimental groups randomly:?1?Control group;?2?DM group;?3?DM plus SC 5 mg/kg group;?4?DM plus 10mg/kg vitexin group;?5?DM plus 20 mg/kg vitexin group;and?6?DM plus 40 mg/kg vitexin group.Mice in the control and experimental groups were administered orally once daily for 62 days.?1?The blood glucose levels were monitored on the 0,10^th,20^th,40^th,and 62^nddays of initializing the treatment throughout the study,and detected the HbAlc levels.?2?The reproductive organ weights were examined.?3?The sperm parameters measured by white blood cell?WBC?chamber by measuring the indexes of sperm counts,sperm motility,sperm viability and the sperm abnormality.?4?Enzyme-linked immunosorbent assay?ELISA?was used to evaluate serum hormonal levels.?5?Oxidative stress indexes were assessed by the method of spectrophotometry in testis tissues.?6?Histopathological impairments were analyzed by and hematoxylin-eosin?HE?staining in diabetic testicular tissues.?7?Ultrastructural changes in diabetic testicular tissues were evaluated by Electron Microscopy.TM3 Leydig cells were induced by high glucose?HG,75mM?for 48h,and were divided into control group,HG group,vitexin?0.1,1,10?g/mL?and LY294002?1?g/mL?group.We used MTT method to evaluate the effect of vitexin on cell viability;Annexin V/PI to examine the effect of vitexin on cell apoptosis rate;JC-1staining to detecte the effect of vitexin on mitochondrial membrane potential?MMP?.2.To explore the relationship between protective effects of vitexin on diabetic testicular dysfunction and PI3K/Akt signaling pathwayWestern blot methods were used to estimate the protein expression of PI3K,Akt,Caspase-3,Bcl-2 and Bax in the teaticular tissue.Western blotmethods were used to estimate the protein expression ofPI3K,p-PI3K,Akt,p-Akt,Caspase-3,Active Caspase-3,Bad,Bcl-2 and Bax in theTM3Leydig cells.ResultsCompared to control group,blood glucose levels and HbAlc levels were significantly higher in the diabetic group?p<0.01,p<0.01,respectively?;meanwhile sperm counts,motility and viability decreased,while sperm abnormality increased?p<0.01,p<0.01,respectively?;serum hormonal levels decreased and the levels of ROS and MDA increased and antioxidant enzyme SOD?GSH-Px and CAT activities decreased in testicular tissues?p<0.01,p<0.01,p<0.01,respectively?.Compared to DM group,there was no significant difference in blood glucose levels at different time points between the groups treated with vitexin?10,20,40 mg/kg?.HbAlc level was significantly decreased?p<0.05,p<0.01,respectively?.Vitexin?20,40 mg/kg?group significantly increased diabetes mice epididymal sperm count,energy and live rate?p<0.05,p<0.01,respectively?,vitexin?40 mg/kg?decrease the rate of sperm abnormality?p<0.01?,vitexin?40 mg/kg?showed a significant increase in diabetes mice serum T,FSH,LH levels?p<0.01?,and can significantly reduced the levels of MDA and ROS in diabetic mice testicular tissue?p<0.01,p<0.01,respectively?,The activity of SOD,GSH-Px and CAT in testicular tissues were increased?p<0.01,p<0.01,respectively?.Vitexin?40 mg/kg?group can significantly reduce the structure damage of testis in diabetic mice,increase the number of leydig cells and reduce the pathological changes of the ultrastructure of testis leydig cells in diabetic mice.Compared to the control group,the cell viability of TM3 Leydig cells in HG group was decreased?p<0.01?,while the intracellular ROS level was increased?p<0.01?,the intracellular MMP was decreased?p<0.01?,and the apoptosis rate was significantly increased?p<0.01?.Compared to HG group,vitexin group?0.1,1,10?g/mL?showed a dose-dependent increase in cell survival rate?p>0.05,p<0.05,p<0.01?,and vitexin group?10?g/mL?showed an increase in MMP level and cell apoptosis rate?p<0.01?.Compared to DM group,vitexin?40 mg/kg?increased the expression levels of PI3K and Akt in testicular tissues?p<0.05,p<0.05,respectively?,reduced the expression of caspase-3 protein?p<0.01?,and significantly increased the expression rate of Bcl-2/Bax protein?p<0.05?.Compared to HG group,the expression levels of PI3K,p-PI3K and p-Akt increased significantly?p<0.05,p<0.05,p<0.05,respectively?,the expression levels of Bcl-2/Bax protein ratio increased significantly?p<0.01,p<0.05,respectively?,and the expression levels of active caspase-3 and Bad protein decreased significantly?p<0.05?after the administration of vitexin?10?g/mL?.Vitexin can be neutralized by LY294002 in regulating the above protein expression partially.Conclusion1.Protective effects of vitexin on diabetic testicular dysfunction in male mice.2.Protective effects of vitexin on diabetic testicular dysfunction in male mice related to activate the Leydig cells PI3K/Akt signaling pathway and decreased Leydig cells apoptosis.