位庚，刘红利，李红蓉，马柳一，尹玉洁，姚兵，贾振华

通心络对同型半胱氨酸诱导大鼠心肌微血管内皮细胞损伤的干预作用及氧化应激机制研究

位庚，刘红利，李红蓉，马柳一，尹玉洁，姚兵，贾振华

目的：探讨通心络对同型半胱氨酸诱导大鼠心肌微血管内皮细胞损伤的干预作用及氧化应激机制。

方法：采用植块法原代培养大鼠心肌微血管内皮细胞，倒置显微镜观察细胞形态并通过CD31免疫荧光法对培养的大鼠心肌微血管内皮细胞进行辨别、鉴定。用同型半胱氨酸（Hcy）建立细胞损伤模型，实验分为对照组、同型半胱氨酸组、通心络低浓度组、通心络中浓度组、通心络高浓度组。分别检测各组细胞的存活率、细胞上清液中超氧化物歧化酶（SOD）活性和丙二醛（MDA）含量、细胞内活性氧（ROS）水平、内皮素-1（ET-1）和内皮型一氧化氮合酶（eNOS）表达水平的变化。

结果：与对照组相比，同型半胱氨酸组细胞存活率降低 ［（74.61±2.88）% vs（100.00±2.07）%］，细胞上清液中MDA含量升高［（4.10±0.18）nmol/ml vs（1.92±0.10）nmol/ml］，SOD活性降低［（7.55±0.71）U/ml vs（20.77±0.68）U/ml］，细胞内ROS水平升高［（38.17±10.28）% vs（19.83±2.97）%］，ET-1 mRNA表达上调，eNOS蛋白表达下调；与同型半胱氨酸组相比，通心络各剂量组均能不同程度的改善上述指标变化。

结论：通心络能够改善同型半胱氨酸诱导的大鼠心肌微血管内皮细胞的功能损伤，并通过减轻氧化应激损伤发挥细胞保护作用。

通心络；心肌微血管内皮细胞；同型半胱氨酸；内皮功能；氧化应激

Abstract

Objective: To observe the effect of Tongxinluo （TXL） on homocysteine-induced rat’s cardiac micro vascular endothelial cell（RCMECs） injury and to study the oxidative stress mechanism.

Methods: Primary RCMECs were cultured with tissue explants process， cell morphology was observed by inverted microscope and the cell was identified by CD31 immunofluorescence method. RCMEC injury model was established by Homocysteine （Hcy）induction and the cells were divided into 5 groups: Control group， with normal cells， Hcy group， the cells were treated by Hcy at 10 mmol/L， Low-dose TXL group， Hcy treated cells were cultured with TXL at 100 mg/L， Middle-dose TXL （200 mg/L）group and high-dose TXL （400 mg/L） group. Cell survival rates were detected， supernatant levels of superoxide dismutase （SOD）activity and malondialdehyde （MDA） content were examined， intracellular protein expressions of reactive oxygen species （ROS）and endothelial nitric oxide synthase （eNOS） were detected and mRNA expression of endothelin-1 （ET-1） was measured in different groups respectively.

Results: Compared with Control group， Hcy group showed decreased cell survival rate （74.61 ± 2.88）% vs （100.00 ± 2.07）%， increased supernatant level of MDA （4.10 ± 0.18） nmol/ml vs （1.92 ± 0.10） nmol/ml， reduced SOD activity （7.55 ± 0.71） U/ml vs（20.77 ± 0.68） U/ml， elevated ROS level（38.17 ± 10.28） % vs （19.83 ± 2.97） %， up-regulated mRNA expression of ET-1 and down-regulated protein expression of eNOS. Compared with Hcy group， the above indexes were improved in each TXL group at different levels.

Conclusion: TXL could decrease Hcy induced RCMECs injury， such protection was conducted by reducing the oxidative stress mechanism in cells.

（Chinese Circulation Journal， 2016，31: 908.）

同型半胱氨酸（Homocysteine，Hcy）是诱发动脉粥样硬化的独立危险因素，它可以通过诱导血管壁氧化应激反应而导致内皮细胞功能障碍［1］，这可能是Hcy导致动脉粥样硬化的机制之一。……