| Objective: To elucidate the effect of Icariin on improving spermatogenic dysfunction of testis in diabetic rats and its molecular mechanism of regulating glycolysis in the testis.Methods: Streptozotocin combined with high-fat diet were used to construct type II diabetic rat model: 8-week-old rats were adaptively fed for 1 week,fasted for 12 hours after high-fat diet for 6 weeks,and 65mg/kg streptozotocin(STZ)was injected intraperitoneally at a time bacterin.After successful modeling,they were divided into control group,diabetes group,Icariin group,and Metformin group,with 12 animals in each group.The Icariin group was administered 80 mg/kg/d Icariin extract,after 6 weeks of treatment,6 rats in each group were randomly selected for weighing,blood glucose and insulin were measured,and the material was obtained.The testis and epididymal tail were collected and weighed,and their index was calculated.The quality parameters of rats semen were determined by blood cell counting plate and eosin staining.HE staining was used to observe the morphological changes of testis,the thickness of spermatogenic epithelium and the diameter of spermatogenic tubules were detected,and the number of spermatogonia cells and sertoli cells were calculated.The expression of PCNA in testis was analyzed by immunohistochemistry,apoptosis of spermatogenic cells was detected by TUNEL,and the expression of apoptosis-related genes was analyzed by QRT-PCR.The changes of glucose metabolites in testis were detected by ELISA.The expression changes of glycolytic rate-limiting enzymes and related regulatory factors were analyzed by QRT-PCR at m RNA and protein level.Results: 1.Weight of diabetic rats was decreased and increased after ICA intervention.2.HE staining results showed that the testicular spermatogenic tubules of type 2 diabetic rats were irregular in morphology,with varying degrees of atrophy,vacuolar-like lesions in the spermatogenic epithelium,sparse and disordered spermatogenic cells,and reduced sperm count.After ICA intervention,the spermatogenic tubules were restored and sperm count increased.3.Immunohistochemical results showed that cell proliferating nuclear antigen(PCNA)was mainly localized in spermatogonia and spermatocyte,and the expression intensity was decreased,and the m RNA expression of PCNA was also down-regulated.After ICA intervention,the expression intensity and level of PCNA were both increased.4.Apoptosis of testicular spermatogenic cells was detected,results showed that spermatogenic cells at all levels in testicular spermatogenic tubule of diabetic rats had different degrees of apoptosis,the number of apoptotic cells increased,the expression of pro-apoptotic genes Bax and Caspase-3 was upregulated,and the expression of anti-apoptotic gene Bcl-2 was downregulated.After ICA intervention,the expressions of Caspase-3 and Bax were down-regulated,and the expression of Bcl-2 was up-regulated,which inhibited the apoptosis of spermatogenic cells.5.After ICA intervention,the lactic acid level in the testis of rats significantly increased,and the expression of the rate-limiting enzyme gene HK2 showed no difference.After ICA intervention,the expression of PKM2 and LDHA were significantly increased,and LDHA was mainly localized in the cytoplasm.6.Immunohistochemical results showed that SIRT1 and HIF-1ɑ were mainly localized in spermatogonial cells and spermatocytes,and their expression intensity decreased with the increase of blood glucose level,and the transcription level was also significantly downregulated.After ICA intervention,the expression of SIRT1 and HIF-1ɑ in rat testis could be up-regulated.Conclusion: Icarin can regulate the testicular glycolysis pathway and inhibit the apoptosis of spermatogenic cells to improve the spermatogenic disorder in type 2 diabetic rats. |
PDF Full Text Request