绞股蓝多糖提取条件优化及对UVB辐射损伤的保护作用

Optimization of Extraction Conditions of Gynostemma Pentaphyllum Polysaccharide and Its Protective Effect Against UVB Radiation Damage

在优化绞股蓝多糖提取条件的基础上,初步评价绞股蓝多糖的抗紫外辐射活性。通过单因素和正交实验确定绞股蓝多糖最佳提取条件,利用人永生化表皮细胞HaCaT制备UVB辐射模型,考察0.25~1.0 mg/mL多糖对细胞存活率的影响,并用DCFH-DA荧光探针标记检测胞内ROS水平,考察多糖对ROS表达水平的影响。结果显示,经单因素和正交实验,绞股蓝多糖最佳提取条件为料液比1:50(w:v)、80℃提取1 h,得率为9.85%。系列质量浓度绞股蓝多糖预处理均可显著改善UVB辐射的HaCaT细胞存活率,1 mg/mL多糖可使120 mJ/cm^(2)剂量下的细胞存活率从29%提高到57.24%。1 mg/mL绞股蓝多糖可显著降低UVB辐射导致的胞内ROS水平,降低UVB所致氧化损伤。研究提示,绞股蓝多糖具有较好的抗UVB辐射活性。

The aim of this study is to optimize the extraction conditions of Gynostemma pentaphyllum polysaccharides,and to evaluate the anti-ultraviolet activity of the polysaccharides.The optimal extraction conditions of gynostemma pentagemma polysaccharides were determined by single factor and orthogonal experiments.The effects of 0.25-1.0mg/mL polysaccharides on cell viability were investigated by UVB radiation model using HaCaT cells.The intracellular ROS level was detected by DCFH-DA fluorescent probe labeling,and the effects of polysaccharides on ROS expression level were investigated.The results showed that the optimal extraction conditions of Gynostemma pentaphyllum polysaccharide were solid to liquid ratio of 1:50(w:v)and 80°C for 1h,and the yield was 9.85%.The survival rate of HaCaT cells treated with UVB radiation was significantly improved by a series of concentration of the polysaccharide pretreatment.The survival rate of HaCaT cells irradiated by 120 mJ/cm^(2) was increased from 29% to 57.24%by 1mg/mL of polysaccharide pretreatment.1mg/mL Gynostemma pentaphyllum polysaccharide can significantly reduce the intracellular ROS level induced by UVB radiation and reduce the UVB induced oxidative damage.The results indicated that Gynostemma pentaphyllum polysaccharide had better anti-UVB radiation activity.