野蔷薇根总黄酮对H2O2诱导损伤的人脐静脉内皮细胞的影响

Effect of Total Flavonoids from Rosa multiflora Thunb on Damage of HUVEC Induced by Hydrogen Peroxide

目的研究野蔷薇根总黄酮(Total Flavonoids from Rosa multiflora Thunb,TF-RM)对过氧化氢(H2O2)诱导损伤的人脐静脉内皮细胞(Human umbilical vein endothelial cells,HUVEC)的影响。方法将SD大鼠随机分为3组,每组10只:空白对照组,维生素C(Vit C)组(0.08 g·kg-1),TF-RM组(2.5 g·kg-1),连续灌胃给药7 d,制备含药血清。采用H2O2诱导HUVEC损伤,分别加入Vit C、TF-RM低、中、高剂量含药血清,噻唑蓝(MTT)法检测细胞活力,AnnexinV-FITC/PI双染法检测细胞凋亡,微量酶标法检测乳酸脱氢酶(LDH)含量,硝酸还原酶法检测一氧化氮(NO)含量,酶联免疫吸附法(ELISA)检测内皮素-1(ET-1)含量。结果与模型组比较,Vit C组、TF-RM低、高剂量组细胞活力明显提高(P<0.05,P<0.01);Vit C组、TF-RM中、高浓度组正常细胞增加,早、晚期凋亡、坏死细胞减少;Vit C组、TF-RM高剂量组LDH含量显著降低(P<0.05,P<0.01);TF-RM高浓度组NO含量显著提高(P<0.05),ET-1含量显著降低(P<0.01)。结论TF-RM对H2O2诱导损伤的血管内皮细胞具有明显保护作用。

Objective To study the effect of total flavonoids from Rosa multiflora Thunb(TF-RM)on human umbilical vein endothelial cells(HUVECs)in vitro.Methods SD rats were randomly divided into blank control group,Vitamin C group(0.08 g·kg-1·d-1)and TF-RM group(2.5 g·kg-1·d-1).After oral administration for seven days,rats were anesthetized to prepare drug-containing serum.Hydrogen peroxide was used to induce damage in HUVEC;and then Vitamin C,serum containing TF-RM of high,medium and low dosages were added to HUVECs.The MTT method was used to detect the cell viability.AnnexinV-FITC/PI double staining was carried out to detect the apoptosis of cells.The micro-enzyme-linked method was applied to detect the release of lactate dehydrogenase(LDH).The nitrate reductase activity method was used to detect the content of nitric oxide.The enzyme-linked immuno sorbent assay was used to detect the content of endothelins-1(ET-1).Results The results revealed that the viability of HUVECs was improved when treated with serum containing Vit C,TF-RM low and high dosage(P<0.05 or P<0.01),respectively.The late stage apoptosis and necrosis of HUVECs was decreased when treated with serum containing VitC,TF-RM medium and high dosage,respectively.The release of LDH was significantly decreased when treated with serum containing VitC and TF-RM high dosage(P<0.05 or P<0.01).The content of nitric oxide was significantly improved(P<0.05)and the content of ET-1 was significantly decreased(P<0.01)when treated with serum containing TF-RM high dosage.Conclusion It is showed that TF-RM has inhibitory effects on damage of HUVECs in atherosclerosis induced by hydrogen peroxide.