摘要
目的探讨北京早园竹叶提取物对H2O2诱导的中国仓鼠卵巢细胞(CHO-K1)DNA损伤的保护与修复作用。方法采用乙醇-水溶液超声辅助提取方法制备竹叶粗提物,经SP825大孔树脂吸附分离,分别以10%,30%,60%的乙醇水溶液洗脱得到不同成分的提取液,并用高效液相色谱(HPLC)定性分析早园竹叶提取物的主要活性成分;采用单细胞凝胶电泳法(SCGE)检测25~400μg/ml H2O2对CHO—K1细胞的DNA损伤作用以及不同组分竹叶提取物在25~50μg/ml浓度下抗H2O2诱导DNA损伤的保护与修复作用。结果竹叶总提物中含有C-黄酮苷类和酚酸类两类活性物质,经SP825大孔树脂纯化后,10%乙醇洗脱成分主要成分为酚酸类物质,30%乙醇洗脱成分中主要为C-黄酮苷类物质。H2O2具有明显的DNA链断裂作用,导致彗星尾长明显增加。全组分、10%组分、30%组分在6.25μg/ml浓度下对H2O2诱导的CHO—K1细胞DNA损伤的修复作用最强,且30%组分的修复作用最明显。在12.5μg/ml下预防H2O2诱导的CHO-K1细胞DNA损伤作用最强,其保护作用均以全组分为明显,由此说明C-黄酮类活性成分在抗遗传毒性作用中起重要作用。结论一定浓度的早园竹叶提取物对H2O2诱导的CHO-K1细胞DNA损伤具有保护与修复作用。
Objective To investigate the preventive and repairing effect of extract of bamboo (North Ph. Praecox) leaves on DNA damage in Chinese hamster ovary cell-K1 (CHO-K1). Methods The crude extracts of North Ph. praecox leaves were prepared by ethanol-aqueous solution with ultrasonic-assisted extraction. The extracts were further purified and isolated by SP825 resin column, then eluted by 10%,30% and 60% ethanol and analyzed qualitatively by HPLC. The DNA damage effect of 25-400 μg/ml H2O2 and the protective and repaired effects of 25-400 μg/ml leaf extracts in different components on CHO-K1 DNA injury by H2O2 were determined by the single cell gel electrophoresis (comet)assay. Results C-glycosides and phenolic acids were detected in crude extracts. After purified by SP825 resin column chromatograph, the main components of 10% ethanol eluent was phenolic acids, and C-glycosides was the main ingredients of 30% ethanol eluent. No C-glycosides and phenolic acids were found in 60% ethanol eluent. H2O2 showed a significant effect on DNA damage, with dose dependent manner, for CHO-K1 cells and DNA tail length showed significant difference, compared with negative control. At concentration of 6.25 μg/ml, crude extracts, 10% ethanol eluent and 30% ethanol eluent had strong repairing DNA damage effect on CHO-K1 cells, and the effect of 30% group was the most obvious. In addition, three groups showed strong anti-DNA damage function to CHO-K1, and protecting effect of crude extracts was the most strong. The results suggest that C-glycosides ingredients play an important role in antigenotoxicity in extract Conclusion The extract of bamboo leaves has an antagonism to the DNA damage induced by H2O2 in CHO-K1.
出处
《环境与健康杂志》
CAS
CSCD
北大核心
2011年第2期118-121,共4页
Journal of Environment and Health
基金
北京市自然科学基金(7092015)
北京联合大学本科生研究计划项目
关键词
竹叶提取物
DNA损伤
单细胞凝胶电泳
H2O2
Extract of bamboo leaves
DNA damage
Single cell gel electrophoresis( SCGE )
H2O2