| ObjectTo observe the protective effect of Astragalin(AG)on spermatogenic disfunction in streptozotocin-induced diabetes in mice,and to explore the mechanism of protective effect of AG on spermatogenic dysfunction in diabetic in mice.Method1.Observing the effect of AG on spermatogenic disfunction in diabetic mice1 Mice were given streptozotocin(STZ)by intraperitoneal injection to build diabetic model,and treated with AG(30 mg/kg,10 mg/kg and 3.3 mg/kg)and Clomiphene(5 mg/kg)in 8 weeks,then,the blood glucose level and HbAlc level were tested in this process.After the administration of AG,the effect of AG on the sperm parameters in diabetic mice was detected by blood cell counting plate;the analytical balance was used to detect organ weight and organ coefficient in diabetic mice;the effect of AG on ALP and ACP in diabetic mice in testicular tissues by spectrophotometry;the effects of AG on the changes of testicular tissue structure in diabetic mice were observed by HE and PAS staining.2 The injured model of GC-1 spermatogonium was induced by high glucose.The effect of AG(10 μg/mL)on apoptosis of GC-1spg was detect by using CCK8 and flow cytometry for 24 h.2.Investigating the protective mechanism of AG on spermatogenic dysfunction in diabetic male mice1 The expression of NF-κB in the testis of diabetic mice was observed by immunohistochemistry;Western blot was used to detect the protein expressions of TNF-α,NF-κB,and iNOS in diabetic testicular tissue in mice.2 Western blot was used to detect the protein expression of IKK-β,p-IKK-β,IκB-α,p-IκB-α,NF-κB,p-NF-κB in HG-induced GC-1spg;the displacement of NF-κB p65 in HG-induced GC-1spg was detect by immunofluorescence.Result1.Effect of AG on spermatogenic disfunction in diabetic mice1 Compared with the diabetic group,the blood glucose level and HbAlc level in AG(30 g/kg)group were decreased at the end of treatment(P<0.05),but did not reach the ormal range;compared with the diabetic group,the AG(30 mg/kg)group was ignificantly increased sperm count,sperm motility,sperm motility and daily sperm production(P<0.01),significantly reduced sperm abnormal rate(P<0.01);AG(30 mg/kg) significantly increased the activity of ALP and ACP in testicular tissue with comparison with diabetic group(P<0.01);compared with diabetic group,AG(30 mg/kg)group can significantly alleviate pathological damage and recovered the basement of testicular seminiferous tubules in diabetic mice.2 Compared with the HG group,the AG(10 μg/mL)group was significantly increased the ell viability of GC-1spg and reduced the apoptosis rate(P<0.01).2.The protective mechanism of AG on spermatogenic disfunction in diabetic male mice1 The results of immunohistochemistry was showed that AG(30 mg/kg)significantly educed the expression of NF-κB p65 in testicular tissue of spermatogenic cells with omparison with diabetic group;simply compared with diabetic group,meanwhile,the esults of Western Blot was showed that the AG(30 mg/kg)group significantly was educed the protein expressions of TNF-α,NF-κB p65,and iNOS in the testis of diabetic ice(P<0.01).2 Compared with HG group,AG(10 μg/mL)group down-regulates the high protein xpressions of p-IKKβ(P<0.05),p-IκBα(P<0.01),p-NF-κB p65(P<0.05);Compared ith HG group,the results of immunohistochemistry was showed that the AG(10 μg/mL) roup can inhibit the migration of NF-κB p65 into the nucleus.Conclusion1.The effect of Astragalin can protect spermatogenic dysfunction in diabetic mice;2.The protective effect of Astragalin on spermatogenic dysfunction in diabetic mice is related to the inhibition of NF-κB signal pathway in spermatogonium. |
