罗布麻总黄酮对过氧化氢致血管内皮细胞损伤的保护作用

摘要

AIM: To study the protective effect of the total flavonoids of Folium Apocyni Veneti (TFF) on human umbilical vein endothelial cells (HUVECs) damaged by H2O2.METHODS: HUVECs were cultured in vitro, and an injured model by H2O2 was established.Cell viability was measured by MTT assay.The rate of apoptosis and cell circles were detected by flow cytometry (FCM).The content of MDA and LDH, the activity of SOD and NO were de- termined by spectrophotometry.The immunohistochemistry was used to observe the expression of NF-kB.RESULTS: Compared with the normal group, the cell viability, the percentage of cells of S phase and G2/M phase, the activity of the cells and the percentage of cells of G0/G1 phase, the level of LDH and MDA along with the expression of NF-kB in was shown to increase cells viability, decrease the content of LDH, MDA and increase the activity of SOD and NO, de- crease the apoptosis rate of cells and the percentage of cells of G0/G1 phase, increase the percentage of cells of S phase and G2/M phase, decrease the expression of NF-kB.CONCLUSION: The total flavonoids of Apocyni veneti, have pre- cise protective effect on HUVECs injury induced by H2O2 which may be related to its antioxidant activity and regulating the expression of NF-kB.%目的:探讨罗布麻总黄酮对过氧化氢(H2O2)所致的人脐静脉血管内皮细胞(HUVEC)损伤的保护作用.方法:采用MTT法观察不同浓度罗布麻总黄酮对HUVEC增殖的影响,流式细胞仪检测不同浓度罗布麻总黄酮对HUVEC细胞周期变化的影响,采用比色法分别检测细胞培养液NO和SOD的活性、MDA和LDH的含量,免疫组化法测定NF-kB的表达.结果:正常细胞组比,200 μmol·L-1 H2O2模型组细胞增殖活性下降,细胞凋亡率明显增加,G0/G1期细胞比率增加,S期细胞和G2/M期细胞比率明显降低,培养上清液中NO和SOD活性明显下降,MDA和LDH的释放量明显增加,NF-kB的表达时明显增加(P＜0.01).不同浓度的罗布麻总黄酮作用后,与模型组比较,可明显提高细胞增殖活性,降低细胞凋亡率,降低G0/G1期细胞比率,增加S期细胞和G2/M期细胞比率;并可提高细胞上清液中NO水平和SOD活性,降低MDA和LDH的释放,可抑制H2O2引起的NF-kB的过度表达.其中,中、高剂量组有显著性差异.结论:罗布麻总黄酮对H2O2引起HUVEC损伤具有明显的保护作用,其作用机制可能与其抗氧化作用及调节有关NF-kB的表述有关.