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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-κB的表达。结果:正常细胞组比,200μmol·L-1H2O2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-κB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-κB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-κB的表达有关。
Objective: To investigate the protective effect of Apocynum venetum flavonoids on the injury of human umbilical vein endothelial cells (HUVECs) induced by hydrogen peroxide (H2O2). Methods: The effects of different concentrations of Apocynum venetum flavonoids on the proliferation of HUVECs were observed by MTT assay. The effects of different concentrations of Apocynum venetum flavonoids on the cell cycle of HUVECs were detected by flow cytometry. The contents of NO and SOD Activity, MDA and LDH levels, and immunohistochemistry to detect the expression of NF-κB. Results: Compared with normal group, the proliferation activity of 200μmol·L-1H2O2 group was decreased, the apoptosis rate was significantly increased, the percentage of cells in G0 / G1 phase increased, the percentage of cells in S phase and G2 / M phase decreased significantly, The activities of NO and SOD in the fluid were significantly decreased, the release of MDA and LDH were significantly increased, and the expression of NF-κB was significantly increased (P <0.01). Compared with the model group, the total flavonoids of Apocynum venetum could significantly increase the cell proliferation, decrease the apoptosis rate, decrease the ratio of G0 / G1 phase cells and increase the ratio of S phase cells and G2 / M phase cells. Increasing NO level and SOD activity in cell supernatant and decreasing the release of MDA and LDH could inhibit the over-expression of NF-κB induced by H2O2. Among them, the medium and high dose groups have significant differences. CONCLUSION: The total flavonoids of Apocynum venetum can protect HUVECs induced by H2O2. Its mechanism may be related to its antioxidation and the regulation of the expression of NF-κB.