山楂核各萃取组分对H_(2)O_(2)诱导氧化损伤细胞保护作用的比较及活性组分筛选

Protection Effect of Hawthorn Kernel Ethanol Extract and Its Derived Fractions on H_(2)_(O) Induced Oxidative Damaged Cells and Active Fractions Screening

评价山楂核乙醇提取物及其各萃取组分对两种氧化损伤细胞的保护作用并筛选活性组分,为其开发利用提供数据支持。以H_(2)O_(2)诱导永生化的人角质形成(Ha Ca T)细胞和人胃黏膜上皮(GES-1)细胞为氧化损伤细胞模型,比较山楂核乙醇提取物(EE)及其各萃取组分对氧化损伤细胞存活率的影响,测定活性组分对氧化损伤细胞内超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)水平的影响,并对活性组分大类成份进行分析。结果显示,EE及其乙酸乙酯萃取组分(EAF)和水萃取组分(WF)对H_(2)O_(2)诱导氧化损伤细胞均具有一定保护作用。其中,25μg/m L EAF可提高氧化损伤Ha Ca T细胞存活率21.23%,使细胞内SOD水平增加9.09%;25μg/m LWF可提高氧化损伤Ha Ca T和GES-1细胞存活率16.20%和23.61%,使细胞内SOD水平分别增加16.62%和17.38%,GSH-Px水平分别增加1.22倍和2.50%。大类成份分析表明,EAF主要含有糖(32.73%)、黄酮(15.70%)和木脂素(8.09%)等;WF主要含有糖(59.59%)和黄酮(1.14%)等。各组分中,WF对H_(2)O_(2)诱导氧化损伤Ha Ca T和GES-1细胞均具有保护作用,效果较好,其主要成份为糖,作用机制可能与调节细胞内抗氧化物酶水平有关,具有较好开发应用价值。

To investigate the protective effect of hawthorn kernel and screen its active fractions on oxidative damaged cells,providing supports for its applications,the oxidative damaged HaCaT cells and GES-1 cells were induced by H_(2)O_(2).MTT assay and biochemical kit were used to detect cell viability and superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)in the oxidation-damaged cells.The chemical components analysis method was used to study the chemical components of the active fractions.The results showed that hawthorn kernel ethanol extract(EE)and its derived fractions got from ethyl acetate(EAF)and water(WF)had protective activity on oxidative damaged cells.25μg/m L of EAF significantly increased the survival rate of oxidation-damaged Ha Ca T cells 21.23%,the levels of SOD in cells were also increased 9.09%.25μg/m L of WF significantly improved HaCaT and GES-1 cells survival rate 16.20%and 23.61%,increased the levels of SOD 16.62%and 17.38%,increased the levels of GSH-Px in cells 1.22 and 2.50%,respectively.The chemical components of ethyl acetate fraction were sugar(32.73%),flavonoids(15.70%),lignans(8.09%)etc.The water fraction contained sugar(59.59%)and flavonoids(1.14%)etc.It was suggested that WF can protect Ha Ca T and GES-1 cells from H_(2)O_(2)induced oxidative damage,and its main component is sugar.The mechanism of WF may be related to the regulation of intracellular antioxidant enzyme levels,which has a good development and application values.