羊栖菜多糖对氧化应激人成纤维细胞的保护作用

Protective effects and mechanisms of Sargassum Fusiforme polysaccharide on human fibroblast after oxidative stress

目的研究羊栖菜多糖对氧化应激的人皮肤成纤维细胞的保护作用及机制。方法以0.8 mmol/L H2O2诱导人皮肤成纤维细胞建立氧化应激模型,MTT法测定不同浓度羊栖菜多糖(15、30、60mg/L)对过氧化氢(H2O2)损伤人皮肤成纤维细胞增殖的影响,Hoechst 33258染色法观察细胞凋亡,流式细胞仪测定细胞周期分布。测定细胞丙二醛(MDA)含量,超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性、还原性谷胱甘肽(GSH)水平和活性氧(ROS)水平。结果羊栖菜多糖可抑制H2O2对人皮肤成纤维细胞增殖活性的下降和凋亡率的上升,维持细胞正常形态。流式细胞仪检测表明,羊栖菜多糖能显著降低H2O2损伤的细胞G0/G1期阻滞现象;升高SOD和GSH-Px活力,降低MDA和ROS水平,升高GSH水平,差异有统计学意义且具有剂量依赖性(P<0.05,P<0.01)。结论羊栖菜多糖能减轻H2O2对人皮肤成纤维细胞的氧化损伤,具有较好的抗氧化保护作用,与其增强抗氧化酶活性,降低脂质过氧化有关。

Objective To study the protective effects of Sargassum Fusiforme polysaccharide（SFPS）on human fibroblast after oxidative stress and its mechanisms.Methods For establishing the oxidatively injured cell model,normal skin fibroblast was incubated with 0.8 mmol/L H2O2.The proliferation of cells treated by different doses of SFPS（15,30 and 60mg/L respectively）was determined by methyl thiazolyl tetrazolium（MTT）assay.Apoptotic rate of the cells was observed using Hoechst 33258 dye under fluorescent microscope and analyzed using flowcytometry.Then the MDA,GSH and ROS levels,the activities of SOD,GSH-Px were measured.Results SFPS could promote fibroblast proliferation and inhibite the raise of cell apoptosis induced by H2O2.It could maintain the normal morphology and decrease the cell cycle blocking in G0/G1 phase of fibroblast after oxidative stress.SFPS with different concentrations could increase the activities of SOD and GSH-Px,increase the level of GSH and decrease the levels of MDA and ROS in a dose dependent manner（P〈0.05,P〈0.01）.Conclusion SFPS could prevent human fibroblast from the oxidative injury induced by hydrogen peroxide.The mechanisms might relate to its enhancement on activities of antioxidase,and decrease of lipid peroxidation.