Geriatrics Research Center, General Hospital of Air Force, PLA, Beijing 100142, China.
Phytomedicine. 2013 Jan 15;20(2):114-9. doi: 10.1016/j.phymed.2012.10.005. Epub 2012 Dec 4.
In the present study, we investigated whether the therapeutic dosages of Ginkgo biloba extract (EGb) and Aspirin (ASP) might synergistically suppress oxidative stress through regulating the expressions of LOX-1 and phosphorylated p38MAPK (p-p38MAPK) in human coronary artery endothelial cells (HCAECs) ex vivo.
HCAECs were stressed with activated platelets (2×10(8)/ml) and followed by ASP (1, 2 or 5 mmol/l), EGb (4, 40 or 400 μg/ml) and combinational (1 mmol/l ASP and 40μg/ml EGb) treatments in three groups for 12 h. Superoxide anion in HCAECs was measured with H2DCF-DA probe. The expressions of LOX-1 and p-p38MAPK were examined by Western blot.
After stimulation of activated platelets, intracellular superoxide anion was increased about 3-folds in HCAECs. Both ASP and EGb reduced superoxide anion in HCAECs in a dosage dependent manner. Combinational administration of ASP and EGb showed synergistic effect. By Western blot analysis, we were able to show that activated platelets markedly enhanced the expressions of LOX-1 and p-p38MAPK. Both ASP and EGb only inhibited LOX-1 expression in a concentration-dependent manner, but not p-p38MAPK. As expected, the combination of ASP and EGb markedly reduced not only the expression of LOX-1 but also the phosphorylation of p38MAPK.
Both EGb and ASP attenuate the oxidative stress of HCAECs stimulated by activated platelets ex vivo. It appears that the synergistic effect of EGb and ASP may correlate with the inhibition of ROS production, LOX-1 expression and p38MAPK phosphorylation.
本研究旨在探讨银杏叶提取物(EGb)和阿司匹林(ASP)的治疗剂量是否通过调节人冠状动脉内皮细胞(HCAEC)中 LOX-1 和磷酸化 p38MAPK(p-p38MAPK)的表达来协同抑制氧化应激。
用激活的血小板(2×10(8)/ml)对 HCAEC 进行应激处理,然后在三组中分别用 ASP(1、2 或 5 mmol/l)、EGb(4、40 或 400 μg/ml)和联合(1 mmol/l ASP 和 40μg/ml EGb)处理 12 小时。用 H2DCF-DA 探针测量 HCAEC 中的超氧阴离子。用 Western blot 检测 LOX-1 和 p-p38MAPK 的表达。
在激活的血小板刺激后,HCAEC 中的细胞内超氧阴离子增加了约 3 倍。ASP 和 EGb 均以剂量依赖的方式减少 HCAEC 中的超氧阴离子。ASP 和 EGb 的联合给药显示出协同作用。通过 Western blot 分析,我们能够表明激活的血小板显着增强了 LOX-1 和 p-p38MAPK 的表达。ASP 和 EGb 仅以浓度依赖的方式抑制 LOX-1 表达,但不抑制 p-p38MAPK。正如预期的那样,ASP 和 EGb 的组合不仅显着降低了 LOX-1 的表达,而且还降低了 p38MAPK 的磷酸化。
EGb 和 ASP 均可减轻体外激活的血小板刺激的 HCAEC 的氧化应激。似乎 EGb 和 ASP 的协同作用可能与抑制 ROS 产生、LOX-1 表达和 p38MAPK 磷酸化有关。