绿豆皮黄酮对H2O2诱导人脐静脉血管内皮细胞损伤的保护作用

Protective Effects of Flavonoids from Mung Bean Hull on Human Umbilical Vein Endothelial Cells Induced by Hydrogen Peroxide

目的:研究绿豆皮黄酮对H2O2诱导损伤人脐静脉血管内皮细胞的保护作用及其机制。方法:用体积分数70%的乙醇提取绿豆皮中黄酮,采用高效液相色谱法测定其有效成分;通过H2O2诱导HUVEC细胞损伤,建立细胞氧化损伤模型。试验被分为正常对照组、H2O2组、维生素C组、绿豆皮黄酮低、中、高剂量组(20,60,80μg/m L),通过MTT法测定细胞增殖能力,生化比色法检测各组细胞及培养液中的超氧化物酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、乳酸脱氢酶(LDH)活性以及谷胱甘肽(GSH)和丙二醛(MDA)含量。结果:H2O2呈剂量依赖性降低内皮细胞活力,并引起细胞凋亡。其中1 000μmol/L H2O2处理HUVEC后,与正常组相比,细胞存活率明显受到抑制,而绿豆皮各剂量组能显著提高细胞和细胞培养液中的GSH-Px、SOD、GSH活性,降低MDA含量,同时降低细胞中LDH活性并增强细胞培养液中的LDH含量。结论:绿豆皮黄酮能抑制H2O2诱导的内皮细胞凋亡,减轻H2O2对内皮细胞的损伤作用,其机制是:绿豆皮黄酮抑制损伤细胞脂质过氧化,提高机体抗氧化酶活性,清除自由基。

Objective: Study on the protective effects and mechanisms of flavonoids from Mung bean hull on human umbilical vein endothelial cells(HUVEC) following oxidative injury caused by hydrogen peroxide(H2O2). Method:Flavonoids were extracted from Mung bean hull with 70% ethanol and its bioactive components were quantified by HPLC.The HUVEC cells oxidative stress damage model was established by using H2O2. Experiments were divided into normal control group, H2O2 injury group, VitC control group, and low, medium, high flavonoids protective groups(20,60,80 μg/mL). The proliferation was determined by the MTT assay. SOD, GSH-Px, LDH activities, GSH, MDA content in HUVEC cells and culture medium were assayed using biochemical colorimetry. Result: H2O2 could decrease the activity of endothelial cells in a dose-dependent manner, and induce apoptosis. Among them, compared with normal control group,cell proliferation was obviously inhibited after 24 h incubation with 1 000 μmol/L H2O2. However, In all dose groups of flavonoids from mung bean hull, the activities of superoxide dismutase(SOD), glutathione peroxidase(GSH-Px) and glutathione(GSH) content were significantly increased(P<0.05). The content of malondialdehyde(MDA) was significantly decreased(P<0.05). The lactate dehydrogenase(LDH) activity was significantly decreased in cells and was significantly reduced in cell culture medium(P <0.05). Conclusion: Flavonoids from mung bean hull can decrease the apoptosis of HUVEC injured by H2O2, its mechanism may be related to inhibiting lipid peroxidation damage cells, effectively improving the cells antioxidant enzyme activity, eliminating free radicals.