Protective effect of HSP27 on Hcy induced vascular endothelial cell injury and its mechanism / 重庆医学

ABSTRACT

Objective To study the protective effect of heat shock protein 27(HSP27)on homocysteine(Hcy)induced vascular endothelial cell injury and its mechanism.Methods Human umbilical vein endothelial cells(HUVECs) were cultured.The influence of Hcy on the survival rate of HUVECs was detected by MTT method;the Annexin V FITC apoptosis detection kit and flow cytometry were used to detect the influence of Hcy and HSP27 on cellular apoptosis;the experiment design adopted the total nitric oxide detection kit(nitrate reductase method) for detecting the NO level in cell culture fluid,the DCFH DA reactive oxygen species (ROS) detection kit and flow cytometry were used to detect intracellular ROS to research possible protection mechanism.Results After action of HUVECs with different concentrations of Hcy,the mean values of survival rates in the 0 mmol/L,0.2 mmol/L,0.4 mmol/L,0.6 mmol/L,0.8 mmol/L and 1.0 mmol/L groups were 97.33％,95.17％,90.72％,78.29％,63.65％ and 41.51％ respectively.The difference between the 0 mmol/L group and 0.2 mmol/L group had no statistical significance(P＞0.05);the survival rate in the Hcy 0.4,0.6,0.8,1.0 mmol/L groups was lower than that in the 0 mmol/L group,the difference was statistically significant(P＜0.05).The cellular survival rate was negatively correlated with the Hcy concentration.The higher the HSP27 concentration,the higher the cellular survival rate,showing the concentration dependence.HSP27 had a protective effect on HUVECs damage.Hcy could induce apoptosis of endothelial cells,while HSP27 could significantly inhibit apoptosis,indicating that HSP27 could reduce the damage of Hcy on endothelial cells.HSP27 could inhibit the NO decrease caused by Hcy and inhibited the production of Hcy induced ROS.Conclusion HSP27 protects Hcyinduced injured endothelial cells by influencing NO expression and regulating intracellular ROS level.