加味补阳还五汤对氧化型低密度脂蛋白损伤血管内皮细胞的保护作用

Protective Effect of Buyang Huanwu Tang on Human Umbilical Vascular Endothelial Cells Injured by Oxidized Low Density Lipoprotein

目的:观察加味补阳还五汤(BHT)含药血清和含药血浆对氧化型低密度脂蛋白(ox-LDL)损伤血管内皮细胞(VEC)的保护作用及机制,同时比较加味BHT含药血清和含药血浆对VEC生成影响的差异。方法:制备加味BHT含药血清和含药血浆,体外培养人脐静脉血管内皮细胞(HUVECs),采用ox-LDL造模,通过噻唑蓝(MTT)法观察细胞活力和Hoechst33258染色法、流式细胞术观察细胞凋亡率,并通过测定一氧化氮(NO),丙二醛(MDA)含量和超氧化物歧化酶(SOD),谷胱甘肽过氧化物酶(GPx)活力,探讨加味BHT含药血清和含药血浆对ox-LDL损伤VEC的保护作用及含药血清和含药血浆之间的差异。结果:与空白组比较,模型组细胞死亡和凋亡率明显升高(P<0.05),和模型组比较,加味BHT含药血清组和含药血浆组能显著降低ox-LDL造成的HUVECs的凋亡和死亡率(P<0.05),细胞活性显著增强(P<0.05),NO含量,SOD和GPx活力显著提高,MDA含量均显著下降(P<0.05)。此外,含药血清组与含药血浆组比较细胞凋亡和死亡率更低(P<0.05),含药血浆组与含药血清组比较产生NO含量明显增加(P<0.05)。结论:加味BHT含药血清和含药血浆对于oxLDL损伤的VEC具有一定保护作用,其机制可能与改善内皮细胞功能、抗氧化损伤、抑制细胞凋亡有关。在一定条件下对于加味BHT含药血浆保护VEC的作用强于含药血清。

Objective： To investigate the protective effects and mechanism of Buyang Huanwu Tang（BHT） on vascular endothelial cells（VEC） injured by oxidized low density lipoprotein（ox-LDL）,and compare the effect between drug-containing serum and drug-containing plasma on VEC. Method： The modified BHTcontaining serum and plasma were prepared. Human umbilical vascular endothelial cells（HUVECs） were treated with serum and plasma containing modified BHT and incubated with ox-LDL for additional 24 hours for ox-LDL modeling. The cell viability was observed by MTT assay,and cells apoptosis rate was observed by using Hoechst33258 staining and flow cytometry; meanwhile the levels of nitric oxide（NO）,malondialdehyde（MDA）,and the activity of superoxidedismutase（SOD）,glutathione peroxidase（GPx） were determined to explore the difference in the protective effect between modified BHT-containing serum and plasma on injured VEC. Result： As compared with the blank control group,the percentage of cells death and apoptosis was significantly higher in the model group（P 0. 05）. As compared with the model group,the percentage of cells death and apoptosis was significantly reduced in modified BHT-containing serum group and plasma group（P 0. 05）; cell viability was significantly increased（P 0. 05）; the levels of NO,SOD and GPx were all significantly increased,the level of MDA was significantly reduced（P 0. 05）. Besides,the percentage of cells death and apoptosis was lower in modified BHTcontaining serum group than that in plasma group（P 0. 05）; however,the level of NO in modified BHTcontaining plasma group was significantly higher than that in serum group（P 0. 05）. Conclusion： The serum and plasma containing modified BHT could prevent VEC from oxidative injury induced by ox-LDL,and the mechanism may be associated with improving the function of endothelial cells,anti-oxidative damage and inhibiting cell apoptosis. The protective effect of modified BHT-containing plasma on VEC was more stronger than the modified BHT-containing serum under certain conditions.