【摘要】 目的 探讨雄蚕益肾方对雄性迟发性性腺功能减退症(LOH)大鼠睾丸组织铁死亡的影响。方法 48只6月龄雄性SD退役种鼠，依照随机数表法分为模型组，雄蚕益肾方低、高剂量组和丙酸睾酮组，每组12只，另设6只8周龄雄性SD大鼠作为对照组。其中，除对照组外的大鼠每日腹腔注射环磷酰胺20 mg/(kg·d)，持续5 d以构建LOH模型;对照组大鼠则每日腹腔注射等体积生理盐水，持续5 d。对照组和模型组给予等体积蒸馏水灌胃，雄蚕益肾方低、高剂量组分别予10.4、41.6 g/(kg·d)雄蚕益肾方浓缩药液灌胃，丙酸睾酮组大鼠肌注5.21 mg/(kg·d)丙酸睾酮，均连续处理28 d。ELISA检测大鼠血清睾酮水平，HE染色和透射电镜观察睾丸组织大体形态及睾丸间质细胞超微结构，RT-qPCR及Western印迹检测睾丸组织铁死亡基因及蛋白表达量。结果 LOH模型大鼠出现生精小管萎缩、结构紊乱、管腔中精子细胞减少等病理改变，睾丸间质细胞核形态改变、线粒体密度增高及嵴减少等超微结构变化;雄蚕益肾方及丙酸睾酮干预后睾丸病理改变及睾丸间质超微结构得到改善。与对照组相比，模型组血清睾酮显著降低(P<0.05);睾丸组织ROS、ACSL4 mRNA及蛋白表达显著升高，FTH1、GPX4和SLC7A11 mRNA及蛋白表达显著降低(P<0.05)。与模型组相比，雄蚕益肾方低、高剂量组及丙酸睾酮组大鼠血清睾酮显著升高(P<0.05)，睾丸组织ROS、ACSL4 mRNA及蛋白表达显著下降(P<0.05);雄蚕益肾方高剂量组FTH1、GPX4、SLC7A11 mRNA及蛋白表达显著升高(P<0.05)。结论 LOH大鼠睾丸组织铁死亡增加，雄蚕益肾方可提升LOH大鼠血清睾酮水平并改善睾丸组织病理改变及超微结构，可能与抑制LOH大鼠睾丸组织铁死亡相关。更多还原

【Abstract】 Objective To explore the effect of Xiongcan Yishen Formula on ferroptosis in testicular tissue of male rats with late-onset hypogonadism( LOH). Methods A total of 48 male Sprague-Dawley rats aged 6 months were randomly divided into model group,low-dose and high-dose Xiongcan Yishen Formula groups and propionate testosterone group,with 12 rats in each group. An additional 6 male Sprague-Dawley rats aged 8 weeks were set as the normal group. Except for the normal group,the rats were intraperitoneally injected with Cyclophosphamide at a dose of 20 mg/( kg·d) for 5 consecutive days to establish the LOH model,while the normal group received an equal volume of physiological saline for 5 days. The normal group and model group were given equal volumes of distilled water by gavage,while the low-dose and high-dose Xiongcan Yishen Formula groups were administered concentrated decoction at doses of 10. 4 g and 41. 6 g/kg/d respectively,and the propionate testosterone group received intramuscular injections of 5. 21 mg/kg/d propionate testosterone,all for 28 consecutive days. ELISA was used to detect serum testosterone levels in rats,HE staining and transmission electron microscopy were used to observe the gross morphology of testicular tissue and the ultrastructure of Leydig cells,and RT-qPCR and Western blotting were used to detect the expression of ferroptosis-related genes and proteins in testicular tissue. Results The LOH model rats exhibited pathological changes such as atrophy of seminiferous tubules,structural disorder,and reduced spermatocytes in the lumen,as well as ultrastructural changes in Leydig cells including altered nuclear morphology,increased mitochondrial density,and reduced cristae. After intervention with Xiongcan Yishen Formula and propionate testosterone,the pathological changes in testis and the ultrastructure of Leydig cells were improved. Compared with the normal group,serum testosterone levels in the model group were significantly decreased( P <0. 05),the expression of ROS,ACSL4 mRNA and protein in testicular tissue was significantly increased,while the expression of FTH1,GPX4,and SLC7A11 mRNA and protein was significantly decreased( P < 0. 05). Compared with the model group,serum testosterone levels in the low-dose and high-dose Xiongcan Yishen Formula groups and the propionate testosterone group were significantly increased( P < 0. 05),and the expression of ROS,ACSL4 mRNA and protein was significantly decreased( P <0. 05); the high-dose Xiongcan Yishen Formula group showed significantly increased expression of FTH1,GPX4,and SLC7 A11 mRNA and protein( P < 0. 05). Conclusion Ferroptosis in testicular tissue is increased in LOH rats,and Xiongcan Yishen Formula can elevate serum testosterone levels and improve pathological changes and ultrastructure in testicular tissue of LOH rats,possibly related to the inhibition of ferroptosis in testicular tissue of LOH rats.更多还原