补阳还五汤精简方对氧化应激损伤血管内皮细胞的保护作用研究

目的观察补阳还五汤精简方对氧化应激损伤后血管内皮细胞的保护作用,并从核转录因子E2相关因子/抗氧化反应元件(NF-E2-related factor 2/antioxidant response element,Nrf2/ARE)途径探讨其作用机制。方法采用H_2O_2作用大鼠血管内皮细胞4 h建立体外血管内皮细胞氧化应激模型,根据不同处理分成正常组,模型组,补阳还五汤含药血清组(补阳组),补阳还五汤精简方含药血清组(5%精简组、10%精简组、20%精简组)6组。采用倒置显微镜观察细胞形态;流式细胞术检测血管内皮细胞调亡;测定细胞SOD活力、MDA含量;采用Western blot法检测细胞Nrf2、血红素加氧酶-1(HO-1)蛋白的表达。结果与模型组比较,含药血清组均能不同程度改善细胞形态,升高细胞存活率,其中10%精简组较5%精简组、20%精简组细胞损伤程度低;与模型组比较,补阳组和10%精简组可有效抑制细胞凋亡(P<0.05);与模型组比较,补阳组及10%精简组可显著提升SOD活力(P<0.01);补阳组、10%精简组及20%精简组可降低MDA含量(P<0.05);补阳组及10%精简组可明显上调Nrf2、HO-1蛋白表达(P<0.05)。结论 10%补阳还五汤精简方可以显著减轻氧化应激造成血管内皮细胞的损伤,抑制细胞调亡,这一作用可能与上调Nrf2/ARE抗氧化信号通路途径表达有关。

Protective Effect of Thin Recipe of Buyang Huanwu Decoction Against Oxidative Stress Injury in Vascular Endothelial Cells

Objective To investigate the protective effect of thin recipe of Buyang Huanwu Decoction on vascular endothelial cells after oxidative stress injury and its mechanism of action based on the nuclear factor-erythroid 2-related factor 2(Nrf2)/antioxidant response element(ARE)pathway.Methods Rat vascular endothelial cells were treated with H2O2 for 4 hours to establish an in vitro model of oxidative stress in vascular endothelial cells,and according to the treatment method,the cells were divided into normal group,model group,serum containing Buyang Huanwu Decoction group(Buyang group),and serum containing thin recipe of Buyang Huanwu Decoction groups(5%,10%,and 20%thin recipe groups).Inverted microscopy was performed to observe cell morphology;flow cytometry was used to measure the apoptosis of vascular endothelial cells;the activity of superoxide dismutase(SOD)and the content of malondialdehyde(MDA)were measured;Western blot was used to measure the protein expression of Nrf2 and heme oxygenase-1(HO-1)in cells.Results Compared with the model group,the drug-containing serum groups had varying degrees of improvement in cell morphology and an increase in cell survival rate,and the 10%thin recipe group had a lower degree of cell injury than the 5%and 20%thin recipe groups.Compared with the model group,the Buyang group and the 10%thin recipe group had significant inhibition of cell apoptosis(P<0.05)and a significant increase in the activity of SOD(P<0.01).The Buyang group,the 10%thin recipe group,and the 20%thin recipe group had a significant reduction in the content of MDA(P<0.05).The Buyang group and the 10%thin recipe group had significant increases in the protein expression of Nrf2 and HO-1(P<0.05).Conclusion The 10%thin recipe of Buyang Huanwu Decoction can significantly reduce oxidative stress injury in vascular endothelial cells and inhibit cell apoptosis,possibly by upregulating the expression of the Nrf2/ARE antioxidant signaling pathway.