五倍子酸对60Co-γ射线辐照人肠上皮细胞的防护效应研究
投稿时间:2018-11-26  修订日期:2019-04-23  点此下载全文
引用本文:孙会娟,赵涛.五倍子酸对60Co-γ射线辐照人肠上皮细胞的防护效应研究[J].药学实践杂志,2019,37(4):299~303
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孙会娟 陕西渭南职业技术学院护理学院 陕西 渭南 714000
空军军医大学军事预防医学系辐射防护医学教研室, 特殊作业环境危害评估与防治教育部重点实验室, 陕西省自由基生物学与医学重点实验室 陕西 西安 710032 
 
赵涛 空军军医大学军事预防医学系辐射防护医学教研室, 特殊作业环境危害评估与防治教育部重点实验室, 陕西省自由基生物学与医学重点实验室 陕西 西安 710032 zhaotao@fmmu.edu.cn 
基金项目:全军重点科研项目(BWS14J033)
中文摘要:目的 研究中药五倍子的有效成分五倍子酸(GA)对电离辐射诱导人肠上皮细胞(HIEC)损伤的防护作用及机制初探。方法 以4 Gy 60Co-γ射线辐照HIEC细胞,建立细胞辐射损伤模型,采用细胞计数试剂盒-8(CCK-8)法观察不同浓度GA分别作用12、24、48 h后的细胞毒作用,以及GA对受照HIEC增殖能力的影响;平板克隆实验,观察GA对受照HIEC克隆形成能力的作用。酶联免疫吸附法(ELISA)检测GA对受照HIEC的超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽(GSH)活力的影响;Western-Blot法检测GA对受照HIEC的G1/S-特异性周期蛋白Cyclin D1和细胞周期素依赖性激酶CDK4的表达的影响,以探讨GA对受照HIEC防护作用机制。结果 研究显示:① GA作用12 h,可以促进HIEC细胞的生长(P<0.05),但随着作用时间的延长以及GA浓度的增加,GA对HIEC细胞的生长表现出一定的毒性作用。② 4Gy的60Co-γ射线照射下,1、10、20 μmol/L的GA预处理能显著促进HIEC的细胞克隆形成率升高(P<0.05)。③相同实验条件下,1、20 μmol/L的GA预处理能显著升高受照HIEC的细胞增殖力(P<0.05)。④同样实验条件下,1、10、20 μmol/L的GA预处理能显著升高HIEC的细胞SOD活力(P<0.05);10 μmol/L的GA预处理能显著升高GSH活力(P<0.05);1 μmol/L的GA预处理能显著升高细胞周期蛋白D1(Cyclin D1)和周期蛋白依赖性激酶4(CDK4)的水平(P<0.05)。结论 本实验条件下GA能一定程度上减轻60Co-γ射线所致HIEC损伤,其作用机制可能与GA降低γ射线引起的脂质过氧化并减轻细胞周期阻滞有关。
中文关键词:五倍子酸  辐射防护  脂质过氧化  细胞周期蛋白D1  周期蛋白依赖性激酶4
 
Protective effects of Gallic acid on ionizing irradiated HIEC cells
Abstract:Objective To investigate the protective effect and primary mechanism of Gallic acid on 60Co-γ ray induced Human Intestinal Epithelial cell (HIEC) damage.Methods HIEC in the exponential phase were inoculated in petri dishes and were randomly divided into sham exposure group and exposure group.The exposure group was exposed to 4Gy 60Co-γ ray.CCK-8 method was used to detect cell proliferation ability.Clone formation method was used to observe the effects on clone formation ability,ELISA assay was used to evaluate the resistance effects on the lipid oxidation reaction,and Western-Blot test was used to observe the effects of GA on cycle protein Cyclin D1 and CDK4 in irradiated HIEC cells.Results 1.GA can promote the growth of HIEC cells 12h after GA treatment (P<0.05).But GA has a toxic effect on the HIEC growth 24h and 48h after the treatment.2.Compared with 4Gy 60Co-γ ray irradiation exposure group,GA(1,10,20 μmol/L) can increase the proliferation rate of HIEC cell significantly (P< 0.05).3.Compared with 4Gy 60Co-γ ray irradiation exposure group,pretreatment with GA(1,20 μmol/L) increased the clone formation rates of HIEC cell significantly (P<0.05).4.Compared with 4Gy 60Co-γ ray irradiation exposure group,SOD activity of GA(1,10,20μmol/L)treatment group increased significantly (P<0.05).GSH activity of 10 μmol/L GA treatment group increased significantly (P<0.05).5.The expression level of cell cycle protein Cyclin D1 and CDK4 in GA (1 μmol/L) treatment group were significantly increased compare with γ ray irradiation exposure group.Conclusion GA has protective effect on 60Co-γ ray irradiated HIEC cells.The mechanism may be related to the GA's capability as antioxidant,scavenging free radicals,regulating cell cycle and inhibiting cell apoptosis.
keywords:gallic acid  radiation protection  lipid oxidation reaction  Cyclin D1  CDK4
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