车叶草苷对少弱精症大鼠抗氧化应激及抗细胞凋亡的作用机制研究  

Mechanism of Asperuloside on Antioxidative Stress and Anti-apoptosis in Rats with Oligoasthenospermia

作　　者：张敏[1] 颜峰 张泰宁 王雪楠[1] ZHANG Min;YAN Feng;ZHANG Taining;WANG Xuanan(Department of Reproductive Medicine,Affiliated Hospital of Jining Medical College,Jining 272029,China;Department of Laboratory Medicine,Qufu Hospital of Traditional Chinese Medicine,Qufu 273100,China)

机构地区：[1]济宁医学院附属医院生殖医学科,济宁272029 [2]曲阜市中医院检验科,曲阜273100

出　　处：《世界中医药》2024年第17期2584-2589,2595,共7页World Chinese Medicine

基　　金：国家自然科学基金项目(82200863);山东省中医药科技项目(Z—2023065);济宁市重点研发计划项目(2023YXNS228)。

摘　　要：目的:探讨车叶草苷(ASP)对少弱精症大鼠抗氧化应激及抗细胞凋亡的作用机制。方法:通过灌胃奥硝唑建立少弱精症大鼠模型,将SD大鼠按照随机数字表法分为对照组、模型组、车叶草苷组(60 mg/kg)和通路抑制剂(第3周腹腔注射2 mg/kg)+车叶草苷组,每组10只。检测大鼠精子参数;酶联免疫吸附试验法检测血清丙二醛(MDA)、谷胱甘肽(GSH)、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)水平;实时萤光定量聚合酶链式反应法检测睾丸组织B细胞淋巴瘤2(Bcl-2)、B细胞淋巴瘤2相关X蛋白(Bax)和核因子E2相关因子2(Nrf2)基因水平;蛋白质免疫印迹法检测睾丸组织Kelch样ECH关联蛋白1(Keap1)、Nrf2和血红素加氧酶1(HO-1)水平。结果:与对照组比较,模型组精子活力、精子浓度、GSH-Px、GSH、SOD、Bcl-2、Keap1、Nrf2、HO-1水平下降(P<0.05),MDA和Bax水平上升(P<0.05);与模型组比较,车叶草苷组精子活力、精子浓度、GSH-Px、GSH、SOD、Bcl-2、Keap1、Nrf2、HO-1水平上升(P<0.05),MDA和Bax水平下降(P<0.05)。结论:ASP能有效改善少弱精症大鼠精子活力、精子浓度,减少睾丸组织细胞的凋亡,改善氧化应激损伤,Keap1/Nrf2信号通路的激活可能是其作用机制。Objective:To investigate the mechanism of asperuloside(ASP)in combating oxidative stress and apoptosis in a rat model of oligoasthenospermia.Methods:A rat model of oligoasthenospermia was established by oral administration of ornidazole.SD rats were randomly divided into a control group,a model group,an ASP group(60 mg/kg),and pathway inhibitor group(intraperitoneal injection at 2 mg/kg in the 3rd week)+ASP group according to a random number table,with 10 rats in each group.Sperm parameters were assessed.Serum levels of malondialdehyde(MDA),glutathione(GSH),superoxide dismutase(SOD),and glutathione peroxidase(GSH-Px)were measured by enzyme-linked immunosorbent assay(ELISA).Real-time quantitative polymerase chain reaction(qPCR)was used to detect gene expression levels of B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax),and nuclear factor erythroid 2-related factor 2(Nrf2)in testicular tissue.Western blot was used to measure levels of Kelch-like ECH-associated protein 1(Keap1),Nrf2,and heme oxygenase-1(HO-1)in testicular tissue.Results:Compared with the control group,the model group showed decreased sperm motility,sperm concentration,GSH-Px,GSH,SOD,Bcl-2,Keap1,Nrf2,and HO-1 levels(P<0.05),and increased MDA and Bax levels(P<0.05).Compared with the model group,the ASP group demonstrated increased sperm motility,sperm concentration,GSH-Px,GSH,SOD,Bcl-2,Keap1,Nrf2,and HO-1 levels(P<0.05),and decreased MDA and Bax levels(P<0.05).Conclusion:ASP can effectively improve sperm motility and concentration in oligoasthenospermia rats,reduce apoptosis in testicular cells,and mitigate oxidative stress damage.Activation of the Keap1/Nrf2 signaling pathway may be the underlying mechanism of action.

关 键 词：车叶草苷 精子活力 精子损伤 Kelch样ECH关联蛋白1/核因子E2相关因子2信号通路 氧化应激 少弱精症 细胞凋亡 性激素 

分 类 号：R285[医药卫生—中药学] R588.1[医药卫生—中医学]