红花水提物不同有效部位的体外抗辐射作用及机制研究  

Radioprotection effects from different extracts of Carthamus tinctorius

作　　者：范莉[1] 刘建香[1] 封江彬[1] 姜晓燕[1] 高刚[1] 田梅[1] 

机构地区：[1]中国疾病预防控制中心辐射防护与核安全医学所、辐射防护与核应急中国疾病预防控制中心重点实验室,北京100088

出　　处：《癌变．畸变．突变》2017年第2期125-128,133,共5页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：北京市自然科学基金项目(7142116)

摘　　要：目的:通过对红花水提物中不同有效部位进行抗辐射效应及作用机制的初步探讨,为开发新型安全有效的辐射防护药物提供实验依据。方法:红花药材用蒸馏水60℃提取得红花水提物,采用大孔树脂HP-20洗脱分离得到水部位和30%、50%、70%、95%醇部位,采用电子自旋共振波谱仪(ESR)检测红花水提物不同有效部位清除自由基的能力;经红花水提物30%及5%醇部位处理后,利用CCK-8法检测其对^(60)Co γ射线照射后人肝L02细胞存活率的影响;流式细胞术研究其对^(60)Co γ射线照射后人淋巴细胞AHH-1细胞凋亡、细胞周期及Bcl-2/Bax比率的影响。结果:ESR自由基清除实验显示红花水提物的不同有效部位均具有清除自由基能力,半数抑制浓度IC 50值为1.7~79.7 mg/m L。CCK-8法检测结果显示30%及50%醇部位处理辐照后L02细胞的存活率分别为128.3%及130.9%,与照射组比较显著增加(P<0.01)。流式细胞术结果显示与照射组凋亡率(24.7%±4.4%)比较,红花水提物30%及50%醇部位处理组凋亡率分别为15.3%±1.6%及15.2%±1.9%,明显降低(P<0.01)。细胞周期实验结果显示与照射组AHH-1细胞G2/M期比例(45.7%±0.7%)比较,红花水提物30%及50%醇部位处理组G2/M期细胞比例分别为(42.3%±0.6%)及(36.4%±0.4%),差异有统计学意义(P<0.01),同时红花水提物30%及50%醇部位能缓解照射引起的Bcl-2/Bax比率下降(P<0.05)。结论:红花水提物的30%及50%醇洗脱部位对^(60)Co γ射线照射的L02、AHH-1细胞具有明显的辐射防护作用,其作用机制可能与清除自由基、改善细胞周期G2/M期阻滞、调控Bcl-2/Bax比率进而抑制细胞凋亡有关,可作为潜在的安全有效的抗辐射药物继续研究。OBJECTIVE：To develop new,safe and effective radioprotectors,the protective effects from different water extracts ofCarthamustinctorius were investigated.METHODS：Safflower water extract was prepared with distilled water at 60 ℃ and isolated by HP-20 macroporous resin. Water elution fraction,30%,50%,70% and 95% ethanol elution fractions were separated for the study. The free radical scavenging ability of the different extraction fractions were evaluated by electron spin resonance （ESR） spectra. CCK-8 assay was performed to evaluate the survival rate of human liver L02 cells in vitro. Flow cytometry was applied to study apoptosis,cell cycle and the ratio of Bcl-2/Bax of 60human lymphocyte AHH-1 after Co ay irradiation.RESULTS：The ESR results showed that all the different effective parts of safflower water extract had the ability to scavenge free radicals,with IC50 at 1.7-79.7 mg/mL. The survival rates from the 30% and 50% ethanol elution were 128.3% and 130.9% by the CCK-8 assay. The survival rates were significantly higher than the irradiated group and suggested that the two parts had equal radioprotection effect to the water extract of safflower. The decreased apoptosis rate of AHH-1 cells： the 30% fraction （15.3%±1.6%） and the 50% fraction（15.2%±1.9%） were significantly lower than that of the irradiation group （24.7%±4.4%） （P〈0.01）. The results of AHH-1 cell cycle analysis showed that the 30% and 50% ethanol elution fractions had significant improvements in the G2/M phase block that was induced by irradiation （P〈0.01）. At the same time,the two fractions increased the Bcl-2/Bax ratio （P〈0.05） and inhibited the cell apoptosis.CONCLUSION：Our data show that the 30% and 50% ethanol elution 60fractions had good radioprotective effects for L02 and AHH-1 cells which were irradiated with Coγrays. The mechanism for the protection may be related to the elimination of free radicals,the improvement of G2/M cell cycle block,and the regulation Bcl-2/Bax ratio in order to inhibit

关 键 词：红花 有效部位 电子自旋共振波谱仪 CCK-8 作用机制 

分 类 号：R345.7[医药卫生—基础医学]