Effect of Tongxinluo on Homocysteine-induced Rat’s Cardiac Micro Vascular Endothelial Cell Injury and the Oxidative Stress Mechanism / 中国循环杂志

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 identiifed by CD31 immunolfuorescence 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.