参杞强精颗粒对小鼠精原细胞氧化损伤的保护作用及其机制  

Protective Effects and Mechanism of Shenqi Qiangjing Granules on H_(2)O_(2)Induced Injury of Mouse Spermatogonia by Inhibiting Oxidative Stress and Ferroptosis

作　　者：甘典辉[1] 夏冰雨 李鑫[2] 赖克道 宾彬[1] 唐爱存[1] GAN Dianhui;XIA Bingyu;LI Xin;LAI Kedao;BIN Bin;TANG Aicun(the First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,China;Graduate School,Guangxi University of Chinese Medicine,Nanning 530200,China;Guangxi Institute of Chinese Medicine and Pharmaceutical Science,Nanning 530022,China)

机构地区：[1]广西中医药大学第一附属医院,南宁530023 [2]广西中医药大学研究生院,南宁530200 [3]广西壮族自治区中医药研究院,南宁530022

出　　处：《医药导报》2025年第3期371-376,共6页Herald of Medicine

基　　金：广西重点研发计划项目(桂科AB22035073);国家自然科学基金资助项目(82160904);广西中医药适宜技术开发与推广项目(GZSY22-18);广西中医药管理局科研课题(GXZYA20230085)。

摘　　要：目的观察参杞强精颗粒含药血清对过氧化氢(H_(2)O_(2))诱导的小鼠精原细胞(GC-1spg)氧化损伤以及铁死亡的保护作用及其机制。方法将健康雄性SD大鼠30只随机分为空白组、左卡尼汀阳性组、参杞强精颗粒组,灌胃给药7 d后,采集含药血清。将小鼠精原细胞随机分为正常对照组、模型对照组、空白血清组、左卡尼汀含药血清组、参杞强精颗粒含药血清组,除正常对照组外,其余各组制备细胞氧化应激损伤模型,各组给药干预24 h,细胞计数试剂-8(CCK-8)法检测细胞存活率,酶联免疫吸附试验(ELISA)法检测细胞活性氧(ROS)、丙二醛(MDA)、过氧化氢酶(CAT)、谷胱甘肽(GSH)、超氧化物歧化酶(SOD)水平;铁离子比色法检测细胞内铁离子水平;比色法检测细胞内Caspase-3与Caspase-9活性;实时荧光定量聚合酶链式反应(qRT-PCR)法测定谷胱甘肽过氧化物酶4(GPX4)、长链酯酰辅酶A合成酶4(ACSL4)mRNA含量。结果与正常对照组比较,模型对照组细胞增殖活性显著下降,细胞内ROS、MDA和铁离子水平显著升高,CAT、GSH、SOD活性显著降低,Caspase-3和Caspase-9活性显著升高,均差异有统计学意义(P<0.05);与模型对照组比较,参杞强精颗粒含药血清组细胞增殖活性显著提高,细胞内ROS、MDA和铁离子水平降低,CAT、GSH、SOD活性升高,Caspase-3和Caspase-9活性降低,均差异有统计学意义(P<0.05);qRT-PCR结果表明,与模型对照组比较,参杞强精颗粒含药血清组GPX4 mRNA表达上调,ACSL4 mRNA表达降低,均差异有统计学意义(P<0.05)。结论参杞强精颗粒对氧化应激损伤和铁死亡的小鼠精原细胞具有显著保护作用,其机制可能与降低Caspase-3和Caspase-9活性及抑制氧化应激损伤和铁死亡有关。Objective To explore the protective effects and mechanism of Shenqi Qiangjing granules containing serum on hydrogen peroxide(H_(2)O_(2))induced oxidative damage and ferroptosis in mouse spermatogonia(GC-1 spg).Methods Thirty SPF grade healthy male SD rats were selected and randomly divided into blank group,levocarnitine group,and Shenqi Qiangjing granules group.After 7 days of intragastric administration,drug-containing serum was collected from each group.Using mouse spermatogonia as a cell model,they were randomly divided into the normal control group,the model control group,blank serum group,levocarnitine containing serum group,and Shenqi Qiangjing granules containing serum group.Except for the normal control group,the other groups used hydrogen peroxide at a concentration of 600μmol·L^(-1) to induce injury to mouse spermatogonia for 4 hours,and then established oxidative stress injury models,after 24 hours of medication intervention in each group.The survival rate of cells was detected using CCK-8 method;The levels of intracellular reactive oxygen species(ROS),malondialdehyde(MDA),catalase(CAT),glutathione(GSH),and superoxide dismutase(SOD)were detected by ELISA;The intracellular iron level was detected by iron ion colorimetry;The activities of Caspase-3 and Caspase-9 were detected by colorimetry;The mRNA levels of glutathione peroxidase 4(GPX4)and ACSL4 were determined by qRT-PCR.Results Compared with the normal control group,the cell proliferation activity of the model control group decreased significantly,the levels of ROS,MDA and Fe 3+were significantly increased,while the activities of CAT,GSH and SOD were significantly decreased in the model control group,however,Caspase-3 and Caspase-9 activities were significantly increased,the results showed significant difference(P<0.05).Compare with the model control group,Shenqi Qiangjing granules containing serum could significantly increase the cell proliferation activity,decrease the levels of ROS,MDA and Fe 3+,increase the activities of CAT,GSH and SOD,and de

关 键 词：参杞强精颗粒 含药血清 小鼠精原细胞 氧化应激损伤 铁死亡 

分 类 号：R286[医药卫生—中药学] R965[医药卫生—中医学]