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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 protective and repair effects of extract from bamboo leaves of early-morning Beijing on DNA damage induced by H2O2 in Chinese hamster ovary cells (CHO-K1). Methods The crude extracts of bamboo leaves were prepared by ultrasonic assisted extraction with ethanol-water solution. The extracts were separated by SP825 macroporous resin and eluted with 10%, 30% and 60% ethanol aqueous solutions respectively. The extracts were extracted with high performance liquid Chromatography (HPLC) qualitative analysis of the early active ingredients of bamboo leaves extract; single cell gel electrophoresis (SCGE) detection of 25 ~ 400μg / ml H2O2 on DNA damage of CHO-K1 cells and the extraction of different components of the leaves Protection and repair of DNA damage induced by H2O2 in the concentration of 25 ~ 50μg / ml. Results The total contents of bamboo leaves contained two kinds of active substances of C-flavonoid glycosides and phenolic acids. After purified by SP825 macroporous resin, the main components of 10% ethanol elution components were phenolic acids, and 30% ethanol elution components In the main C-flavonoid glycosides. H2O2 has obvious DNA strand breaks, leading to a significant increase in tail length of comet. All the components, 10%, 30% components at 6.25μg / ml H2O2-induced DNA damage repair the strongest role of CHO-K1 cells, and 30% of the components of the most obvious repair. Pretreatment with 12.5μg / ml H2O2 induced the highest DNA damage in CHO-K1 cells. The protective effect of all the components was obvious, indicating that C-flavonoids play an important role in the anti-genotoxic effects. Conclusion A certain concentration of the early leaves extract of bamboo leaves of CHO-K1 cells induced by H2O2 can protect and repair DNA damage.