薯蓣皂苷元对小鼠免疫性睾丸炎的作用及机制

Effect and mechanism of diosgenin on autoimmune orchitis in mice

目的:研究薯蓣皂苷元(diosgenin, Dio)对小鼠免疫性睾丸炎的作用及机制。方法:小鼠按随机数字表法分为空白组、模型组、盐酸左氧氟沙星组,以及Dio低、高剂量组,左侧睾丸注射30μL 10%冰醋酸诱导小鼠免疫性睾丸炎。给药21 d后,检测睾丸组织氧化应激及炎性因子水平、病理变化、细胞凋亡情况、NF-κB/NLRP3和Keap/Nrf2信号通路相关蛋白及阳性细胞表达。结果:模型组小鼠睾丸部分生精上皮结构紊乱,出现空泡样变,睾丸间质出现大量炎性浸润;Dio组小鼠睾丸各级病理改变得到有效恢复。与空白组比较,模型组小鼠血清IL-1β、TNF-α、IL-6和MDA水平明显升高(P<0.01),SOD和GSH-Px水平明显降低(P<0.01),睾丸细胞凋亡率明显升高(P<0.01),睾丸NF-κB、NLRP3、ASC、Caspase-1、IL-1β、Keap1、Nrf2蛋白及NF-κB、Nrf2阳性细胞表达明显升高(P<0.05,P<0.01)。与模型组比较,Dio组上述炎症及氧化应激指标水平明显纠正(P<0.01),睾丸细胞凋亡率明显降低(P<0.01),睾丸Nrf2蛋白及阳性细胞表达明显升高(P<0.01),其余蛋白及阳性细胞表达均明显降低(P<0.01)。结论:Dio可改善免疫性睾丸炎小鼠的炎症及氧化应激,机制可能与调控NF-κB/NLRP3和Keap/Nrf2信号通路有关。

OBJECTIVE To investigate the effects and mechanisms of diosgenin(Dio)on autoimmune orchitis in mice.METHODS Mice were randomly divided into the control group,model group,levofloxacin hydrochloride group,and low-and high-dose Dio group.Immune orchitis was induced by injecting 30μL of 10%glacial acetic acid into the left testis in mice.After 21 days of administration,the levels of oxidative stress and inflammatory factors in testicular tissue,pathological changes,cellular apoptosis,and the expression of proteins associated with the NF-κB/NLRP3 and Keap/Nrf2 signaling pathways,as well as positive cell expression were examined.RESULTS In the model group,the seminiferous epithelial structure of the testes was disrupted,vacuolation occurred,and the testicular interstitium showed extensive inflammatory infiltration.In the Dio groups,the testicular pathological changes were effectively restored.Compared with the control group,the model group exhibited a significant increase in serum levels of IL-1β,TNF-α,IL-6 and MDA(P<0.01),and a significant decrease in SOD and GSH-Px levels(P<0.01);the apoptotic rate markedly elevated;the protein expression of NF-κB,NLRP3,ASC,Caspase-1,IL-1β,Keap1,Nrf2,as well as the positive cell expression of Nrf2 and NF-κB,were significantly upregulated(P<0.05,P<0.01).Compared with the model group,the Dio group exhibited significant correction in the levels of inflammatory and oxidative stress markers mentioned above(P<0.01);the apoptosis rate of testicular cells significantly decreased(P<0.01);the expression of Nrf2 protein and positive cells in the testes significantly increased(P<0.01),while the expression of other proteins and positive cells significantly decreased(P<0.01).CONCLUSION Dio can improve inflammatory and oxidative stress in immune orchitis mice,potentially through regulating the NF-κB/NLRP3 and Keap/Nrf2 signaling pathways.