金松双黄酮对砷致大鼠海马神经细胞衰老的干预效应  

Effect of sciadopitysin on sodium arsenite-induced senescence of rat hippocampal neurons

作　　者：陈诚 陈雄[1] 张爱华[1] CHEN Cheng;CHEN Xiong;ZHANG Aihua(Key Laboratory of Environmental Pollution Monitoring and Disease Control,Ministry of Education/School of Public Health,Guizhou Medical University,Guiyang,Guizhou 550025,China)

机构地区：[1]贵州医科大学,环境污染与疾病监控教育部重点实验室/公共卫生与健康学院,贵州贵阳550025

出　　处：《环境与职业医学》2022年第5期550-555,共6页Journal of Environmental and Occupational Medicine

基　　金：喀斯特地区特色民族医药若干基础问题研究项目(U1812403)。

摘　　要：[背景]长期砷暴露人群除表现出典型的皮肤损害体征外,往往还伴随神经行为异常症状和体征。[目的]探讨金松双黄酮对亚砷酸钠致大鼠神经细胞衰老的干预效应以及细胞周期相关转录因子E2F1在其中可能的介导作用。[方法]采用4μmol·L亚砷酸钠作用于SH-SY5Y细胞24 h,并分别使用50μg·mL^(-1)银杏叶提取物EGb761及四种主要银杏叶双黄酮(异银杏双黄酮、7-去甲基银杏双黄酮、金松双黄酮和银杏双黄酮)干预24 h,采用CCK-8法测定细胞活性。将32只180~200 g SPF级大鼠随机分为对照组、染砷组(10 mg·kg^(-1))、银杏叶提取物干预组(10 mg·kg^(-1))和金松双黄酮干预组(10 mg·kg^(-1)),每组8只,雌雄各半。采用自由饮水方式进行染毒,连续染毒3个月;干预在染毒2个月后进行,采用灌胃的方式进行给药,干预1个月。采用HE染色法检测大鼠海马结构的改变,采用尼氏染色法检测海马形态及神经细胞数量的改变。此外,分别采用β-半乳糖苷酶(SA-β-gal)染色法和Western blotting法检测海马神经细胞的衰老情况及E2F1的表达水平。[结果]与染砷组相比,银杏叶提取物和四种主要的银杏叶双黄酮均有不同程度的细胞活性恢复作用(均P <0.05),其中金松双黄酮相对于银杏叶提取物在拮抗亚砷酸钠神经细胞毒性方面具有更好的活性恢复作用(P <0.05)。HE染色和尼氏染色结果表明:与对照组相比,染砷组大鼠海马神经细胞明显减少且突触结构异常,细胞发生肿胀、核皱缩,出现空泡现象;与染砷组大鼠比较,银杏叶提取物干预组和金松双黄酮干预组大鼠海马神经细胞明显增多且突触结构较正常。SA-β-gal染色结果表明:与对照组(2.88±0.84)相比,染砷组(15.75±3.01)的衰老细胞数量明显增加(P <0.05);与染砷组相比,银杏叶提取物干预组(9.38±1.92)和金松双黄酮干预组(7.75±2.38)衰老细胞数量明显下降(均P <0.05)。Western blotting结果表明:�[Background] In addition to the typical signs of skin damage, long-term arsenic exposure is often accompanied by signs and symptoms of neurobehavioral abnormalities.[Objective] To investigate potential intervention effect of sciadopitysin on senescence of neurons induced by sodium arsenite in rats and possible underlying mediating effect of cell cyclerelated transcription factor E2F1.[Methods] SH-SY5 Y cells were treated with 4 μmol·Lsodium arsenite for 24 h and intervened with 50 μg·m LGinkgo biloba extract(EGb761) or four major biflavonoids in Ginkgo biloba leaves(isoginkgetin, bilobetin, sciadopitysin,and ginkgetin) for 24 h respectively. Then, cell viability was measured by CCK-8 assay. Thirty-two 180-200 g SPF rats were randomly divided into a control group, an arsenic treatment group(10 mg·kg^(-1)), a Ginkgo biloba extract intervention group(10 mg·kg^(-1)), and a sciadopitysin intervention group(10 mg·kg^(-1)), 8 rats in each group, half male and half female. The rats were treated with sodium arsenite by free drinking water for 3 consecutive months, and the intervention treatment was conducted after 2 months of poisoning with drug intake by gavage for 1 month. HE staining was used to detect structural changes in the hippocampus, while Nissl’s staining was used to detect changes in hippocampal morphology and neuron numbers. Moreover, senescence-associated β galactosidase(SA-β-gal) staining and Western blotting were used to detect senescence of hippocampal neurons and the expression level of E2F1, respectively.[Results] Compared to the arsenic treatment group, EGb761 and the four biflavonoids in Ginkgo biloba leaves effectively antagonized the inhibitory effect of sodium arsenite on cell viability(all Ps < 0.05), and sciadopitysin showed better restoration of cellular viability than Ginkgo biloba extract(P < 0.05). The results of HE staining and Nissl’s staining showed that the hippocampal neurons in the arsenic treatment group were reduced in cell count and the synaptic structure was abnormal, w

关 键 词：亚砷酸钠 金松双黄酮 海马 神经细胞衰老 

分 类 号：R114[医药卫生—卫生毒理学]