淫羊藿苷调控P53/SLC7A11/GPX4通路抑制睾丸细胞铁死亡改善少弱精子症小鼠生精功能  

Icariin Improves Spermatogenic Function in Mice with Oligospermia through Inhibiting Testicular Ferroptosis via Regulating P53/SLC7A11/GPX4 Pathway

作　　者：江晓翠 萧闵[1,2] 刘祺 赵敏[2,4] 吕银娟 陈思易 曹继刚 JIANG Xiao-cui;XIAO Min;LIU Qi;ZHAO Min;LÜYin-juan;CHEN Si-yi;CAO Ji-gang(Traditional Chinese Medicine Experimental Center,Hubei University of Chinese Medicine,Wuhan,430065;Hubei Shizhen Laboratory,Wuhan,430065;Department of Urology,Hubei Hospital of Traditional Chinese Medicine,Wuhan,430061;College of Basic Medical Sciences,Hubei University of Chinese Medicine,Wuhan,430065)

机构地区：[1]湖北中医药大学中医药实验中心,武汉430065 [2]湖北时珍实验室,武汉430065 [3]湖北省中医院泌尿外科,武汉430061 [4]湖北中医药大学基础医学院,武汉430065

出　　处：《中国中西医结合杂志》2024年第11期1372-1379,共8页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金青年项目(No.82104704);湖北省中医药管理局中医药重点项目(No.ZY2023Z024);湖北省中医药管理局中医药指导性项目(No.ZY2023F134);武汉中青年名中医项目(No.武卫[2019]17号);湖北中医药大学校级科研平台项目(No.中医校[2022]80号);湖北省教育厅科研计划项目(No.D20222006)。

摘　　要：目的 观察淫羊藿苷通过肿瘤抑制因子(P53)/溶质载体家族7成员11(SLC7A11)/谷胱甘肽过氧化物酶4(GPX4)通路改善小鼠少弱精子症的作用。方法 将KM小鼠随机分为空白组、模型组、淫羊藿苷高、中、低(117.0、58.5、29.3 mg/kg)剂量组和左卡尼汀(400 mg/kg)组,每组10只。除空白组外,采用腺嘌呤(1.25 g/kg)灌胃14天构建少弱精子症模型后,给予相应剂量药物连续灌胃21 d。动物精子分析仪检测精子质量;ELISA检测血清睾酮(T)、雌二醇(E_(2))、促卵泡激素(FSH)水平;HE染色评估睾丸生精功能;普鲁士蓝染色法评估睾丸组织铁沉积;免疫组化法评估睾丸组织SLC7A11、GPX4蛋白表达量;生化法检测睾丸组织超氧化物歧化酶(SOD)活性及丙二醛(MDA)含量;Western Blot及RTPCR检测睾丸组织P53、GPX4、SLC7A11通路蛋白及m RNA表达。结果 模型组小鼠曲细精管分布疏松,生精细胞排列紊乱,管腔内仅见少量精子;各干预组干预后,睾丸形态明显改善,生精细胞不同程度增多,管腔内可见较多游离精子。与空白组比较,模型组小鼠精子密度和活动率、血清T、E_(2)水平、睾丸生精功能总分(TMS)、睾丸组织SLC7A11、GPX4蛋白及m RNA表达、SOD活性降低(P<0.01),血清FSH水平、铁沉积阳性平均光密度(AOD)、睾丸组织MDA含量、P53蛋白及m RNA表达升高(P<0.01)。与模型组比较,干预组小鼠血清T水平、睾丸组织GPX4蛋白及m RNA表达、P53 mRNA表达、SOD活性升高,血清FSH水平降低(P<0.05,P<0.01);除淫羊藿苷低剂量组外其他干预组小鼠精子密度和活动率、血清E_(2)水平、TMS、睾丸组织SLC7A11蛋白及m RNA表达升高(P<0.05,P<0.01),铁沉积阳性AOD、睾丸组织P53蛋白表达降低(P<0.05,P<0.01);淫羊藿高、中剂量组小鼠睾丸组织SLC7A11蛋白表达升高(P<0.05);高剂量组及左卡尼汀组小鼠睾丸组织MDA含量降低(P<0.05)。结论 淫羊藿苷改善少弱精子症小鼠睾丸生精功能的机制可能与调控Objective To investigate the effect and mechanism of icariin in improving oligospermia mice through tumor suppressor factor(P53)/solute carrier family 7 member 11(SLC7A11)/glutathione peroxidase 4 (GPX4) pathway. Methods KM mice were randomly divided into blank group,model group,high,medium,low icariin (117.0,58.5,29.3 mg·kg-1) and levocarnitine (400 mg·kg-1) group,with 10 mice in each group.Except for the blank group,adenine (1.25 g·kg-1) was administered intragastrically for 14 days to establisha model of oligoasthenospermia,and the corresponding dose of drug was administered intragastrically for21 days. Sperm quality was detected by animal sperm analyzer. ELISA was used to detect serum testosterone(T),estradiol (E_(2)) and follicle-stimulating hormone (FSH) levels. HE was used to evaluate testicular spermatogenicfunction. Prussian blue staining was used to evaluate testicular iron deposition. Immunohistochemistry was usedto evaluate testicular tissue SLC7A11 and GPX4 protein expression. Biochemical analysis was used to detectthe activity of superoxide dismutase(SOD) and malondialdehyde(MDA) content in testicular tissue. WesternBlot and RT-PCR was used to detect the protein and mRNA expression of P53,GPX4,SLC7A11 in testiculartissues,respectively. Results In the model group,the distribution of seminiferous tubules was loose,andthe arrangement of spermatogenic cells was disordered,and only a small number of sperm could be seen in thelumen. After the intervention in each intervention group,the morphology of the testes was significantly improved,the spermatogenic cells increased to varying degrees,and more free sperm could be observed in the lumen.Compared with the blank group,the sperm density and motility rate,serum T and E_(2) levels,total testicularspermatogenic function score (TMS),testicular tissue SLC7A11,GPX4 protein and mRNA expression,andSOD activity decreased(P<0.01),while serum FSH level,the positive average optical density(AOD) of irondeposition,MDA content in testicular tissue,and P53 protein and

关 键 词：淫羊藿苷 少弱精子症 肿瘤抑制因子 溶质载体家族7成员11 谷胱甘肽过氧化物酶4 铁死亡 睾丸生精功能 

分 类 号：R285.5[医药卫生—中药学]