目的 探讨葡萄籽原花青素(GSPs)对砷暴露小鼠睾丸核因子-E2相关因子(Nrf2)抗氧化系统和线粒体生物合成的影响.方法 ICR小鼠随机分为对照组、模型组、GSPs组和实验组.模型组和实验组小鼠饮用亚砷酸钠水溶液(10 mg·L-1砷)诱导睾丸损伤,GSPs组和实验组给予100 mg·kg GSPs灌胃,对照组和模型组给予等体积蒸馏水灌胃,每天1次,连续8周.用苏木精-伊红(HE)染色制片对睾丸组织学参数测量,用试剂盒法测定睾丸组织丙二醛(MDA)、超氧化物歧化酶(SOD)和腺苷三磷酸(ATP)含量,用实时定量逆转录聚合酶链反应(RT-PCR)法检测血红素加氧酶1(HO-1)、醌氧化还原酶1(NQO1)的表达水平,用免疫组化法检测睾丸组织Nrf2蛋白表达,用蛋白质印迹法检测睾丸线粒体合成相关蛋白的表达.结果 对照组、模型组、GSPs组、实验组的生精小管直径分别为(184.32±14.14)、(170.41±10.70)、(186.87±8.03)和(181.70±9.15)μm,MDA 分别为(2.30±0.26)、(3.28±0.64)、(2.32±0.40)和(2.74±0.31)nmol·mg-1,Nrf2 阳性表达细胞比例分别 为(46.50±11.98)％、(22.33±8.82)％、(51.67±12.44)％和(39.83±8.35)％,HO-1 mRNA 表达水平分别为 1.00±0.21、0.51±0.10、1.00±0.28 和 0.80±0.06,NQO1 mRNA 表达水平分别为 1.00±0.18、0.59±0.11、1.09±0.28 和 0.81±0.08,ATP 分别为(491.83±67.16)、(368.81±69.93)、(512.44±70.96)和(472.20±68.24)μmol·g-1,PGC-1α蛋白相对表达水平分别为 1.00±0.06、0.22±0.03、0.94±0.05 和 0.48±0.05,TFAM蛋白相对表达水平分别为1.00±0.07、0.32±0.05、0.80±0.05和0.67±0.06.实验组的上述指标与模型组相比,在统计学上差异均有统计学意义(P＜0.05,P＜0.01).结论 GSPs可通过激活Nfr2抗氧化系统和促进线粒体生物合成,对砷诱导的小鼠睾丸损伤具有保护作用.
Objective To investigate the effect of grape seed proanthocyanidins(GSPs)on nuclear factor erythroid 2-related factor 2(Nrf2)antioxidant system and mitochondrial biosynthesis in testes of arsenic-exposed mice.Methods ICR mice were randomly divided into four groups:Control group,model group,GSPs group and experimental group.The mice in the model group and the experimental group drank sodium arsenite solution(10 mg·L-1 arsenic)to induce testicular injury.GSPs group and experimental group were given 100 mg·kg-1 GSPs by gavage,control group and model group were given equal volume of distilled water by gavage once a day for 8 weeks.The histological parameters of testes were measured by hematoxylin-eosin(HE)staining.The contents of malondialdehyde(MDA),superoxide dismutase(SOD)and adenosine triphosphate(ATP)in testicular tissue were determined by kit method.The expression levels of heme oxygenase 1(HO-1)and quinone oxidoreductase 1(NQ01)were detected by real-time quantitative reverse transcription polymerase chain reaction(RT-PCR).The expression of Nrf2 protein in testicular tissue was detected by immunohistochemistry.The expressions of mitochondrial synthesis-related proteins were detected by Western blot.Results The diameters of seminiferous tubules in control group,model group,GSPs group and experimental group were(184.32±14.14),(170.41±10.70),(186.87±8.03)and(181.70±9.15)μm;the contents of MDA were(2.30±0.26),(3.28±0.64),(2.32±0.40)and(2.74±0.31)nmol·mg-1;the proportions of Nrf2 positive cells were(46.50±11.98)％,(22.33±8.82)％,(51.67±12.44)％and(39.83±8.35)％;the mRNA expression levels of HO-1 were 1.00±0.21,0.51±0.10,1.00±0.28 and 0.80±0.06;the mRNA expression levels of NQO1 were 1.00±0.18,0.59±0.11,1.09±0.28 and 0.81±0.08;the contents of ATP were(491.83±67.16),(368.81±69.93),(512.44±70.96)and(472.20±68.24)μmol·g-1;the relative expression levels of PGC-1α protein were 1.00±0.06,0.22±0.03,0.94±0.05 and 0.48±0.05;the relative expression levels of TFAM were 1.00±0.07,0.32±0.05,0.80±0.05 and 0.67±0.06.The above indexes in the experimental group were statistically significant compared with the model group(all P＜0.05,P＜0.01).Conclusion GSPs has a protective effect against arsenic-induced testicular injury in mice by activating the Nfr2 antioxidant system and promoting mitochondrial biosynthesis.
曾群;赵果毅;吴娜;杨李阳;苗宇船
山西中医药大学 基础医学院,山西 晋中 030619
中医学
葡萄籽原花青素;砷;氧化应激;核因子NF-E2相关因子;线粒体生物合成
grape seed proanthocyanidins;arsenic;oxidative stress;nuclear factor erythroid 2-related factor 2;mitochondrial biogenesis
《中国临床药理学杂志》 2024 (005)
713-717 / 5
山西省基础研究计划基金资助项目(20210302123232);山西省卫生健康委科研基金资助项目(2020097);山西中医药大学科技创新能力培育计划基金资助项目(2020PY-JC-08);山西中医药大学山西教育厅基金资助项目(2020L0459);山西中医药大学博士科研启动基金项目(2023BK55)
10.13699/j.cnki.1001-6821.2024.05.017
评论