二陈汤合桃红四物汤含药血清对ox-LDL诱导内皮细胞损伤的保护作用及机制

Effect and mechanism of serum containing of Erchen decoction and Taohong Siwu decoction on ox-LDL induced endothelial cell damage

目的观察二陈汤合桃红四物汤含药血清对氧化型低密度脂蛋白(ox-LDL)诱导内皮细胞损伤的保护作用并探讨其可能的机制。方法培养EA.hy926细胞,随机分为对照组、ox-LDL组、干预组、抑制剂组。流式细胞仪检测各组细胞凋亡水平,比色法检测细胞内丙二醛(MDA)、一氧化氮(NO)水平及超氧化物歧化酶(SOD)活性,ELISA检测细胞上清液中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)水平,Western blot检测磷脂酰肌醇3-激酶(PI3K)、磷酸化AKT(p-AKT)、内皮型一氧化氮合酶(eNOS)蛋白表达水平。结果与对照组比较,ox-LDL组细胞上清液中TNF-α和IL-6水平显著升高(P<0.01),细胞内MDA含量显著升高(P<0.01),SOD活性明显降低(P<0.05),上清液中NO水平显著降低(P<0.01),活性氧(ROS)含量和细胞凋亡率明显升高,PI3K、eNOS蛋白表达水平和AKT蛋白磷酸化水平显著降低(P<0.01)。与ox-LDL组比较,干预组细胞上清液中TNF-α和IL-6水平显著降低(P<0.01),细胞内MDA含量显著降低(P<0.01),SOD活性明显升高(P<0.05),上清液中NO水平显著升高(P<0.01),ROS含量和细胞凋亡率明显降低,PI3K、eNOS蛋白表达水平和AKT蛋白磷酸化水平显著升高(P <0.01)。与干预组比较,抑制剂组细胞上清液中TNF-α和IL-6水平显著升高(P<0.01),细胞内MDA含量显著升高(P<0.01)、SOD活性明显降低(P<0.05),上清液中NO水平明显降低(P<0.05),细胞内ROS含量升高,PI3K、eNOS蛋白表达水平和AKT蛋白磷酸化水平显著降低(P<0.01)。结论二陈汤合桃红四物汤含药血清能有效保护ox-LDL诱导的内皮细胞损伤,其机制可能与激活PI3K/AKT/eNOS信号通路有关。

Aim To investigate the effect and mechanism of Erchen decoction and Taohong Siwu decoction containing serum on oxidized low density lipoprotein( ox-LDL) induced endothelial cell damage. Methods EA. hy926 cells were cultured in vitro,then randomly divided into control group,ox-LDL group,intervention group,inhibitor group.Cell apoptosis was detected by flow cytometry,malondialdehyde( MDA),nitric oxide( NO) and superoxide dismutase( SOD) activity were detected by colorimetry,tumor necrosis factor-α( TNF-α) and interleukin-6( IL-6) levels in cell supernatant were detected by ELISA,and phosphatidylinositol 3-kinase( PI3 K),phosphorylated AKT( p-AKT) and endothelial nitric oxide synthase( eNOS) protein expressions were detected by Western blot. Results Compared with control group,the level of TNF-α and IL-6 increased significantly( P<0.01),MDA content in cells increased significantly( P<0.01),SOD activity decreased( P<0.05),NO level in supernatant decreased significantly( P<0.01),content of reactive oxygen species( ROS) and apoptosis rate increased significantly,the protein expression level of PI3 K,eNOS and the protein phosphorylation of AKT were significantly decreased( P<0.01) in ox-LDL group. Compared with ox-LDL group,the level of TNF-α and IL-6 in cell supernatant decreased significantly( P<0.01),MDA content in cells decreased significantly( P<0.01),SOD activity increased significantly( P<0.05),NO level in the supernatant increased significantly( P<0.01),ROS content and apoptosis rate decreased significantly,the protein expression level of PI3 K,eNOS and the protein phosphorylation of AKT were significantly increased( P<0.01) in intervention group. Compared with intervention group,the level of TNF-α and IL-6 increased significantly( P<0.01),MDA content in cells increased significantly( P<0.01),SOD activity decreased( P<0.05),NO level in supernatant decreased significantly( P< 0.05),content of ROS and apoptosis rate increased significantly,the protein expression level of PI3 K,eNOS and the protein phosphorylation of AKT were significantly decreased( P<0.01) in inhibitor group. Conclusion The serum containing of Erchen decoction and Taohong Siwu decoction can effectively protect ox-LDL induced endothelial cell injury,which may be related to activation of PI3 K/AKT/eNOS signaling pathway.