目的 研究瑞舒伐他汀（Rosu）对同型半胱氨酸（Hcy）诱导的内皮祖细胞（EPCs）活性氧（ROS）产生以及凋亡的影响。方法 从外周血中分离EPCs与Hcy共孵育，或经Rosu、不同的应激信号通路抑制剂甲羟戊酸（100 μmol/L）、乙酰半胱氨酸（NAC，10 μmol/L）、NADPH氧化酶（Nox）抑制剂（DPI，10 μmol/L）、内皮一氧化氮合成酶抑制剂（LNMA，1 mmol/L）预培养后再加入Hcy共孵育。采用流式法检测细胞凋亡率，2,7-二氯荧光黄双乙酸盐法（H2DCF-DA）检测细胞内ROS水平，光泽精化学发光法检测Nox活性，RT-PCR检测Nox4 mRNA的表达。结果 Rosu显著抑制了Hcy诱导的ROS蓄积和EPCs凋亡，拮抗了Hcy诱导的Nox激活以及Nox4 mRNA表达。Nox4 siRNA转染EPCs可以产生相似的效果。结论 Rosu对EPCs的保护作用可能是通过Nox4途径抑制Hcy诱导的Nox激活、ROS蓄积和EPCs凋亡来实现。

Objective To study inhibition of rosuvastatin (Rosu) on homocysteine (Hcy)-induced reactive oxygen species (ROS) and apoptosis in endothelial progenitor cells (EPCs). Methods EPCs were isolated from peripheral blood and then incubated with Hcy, or pre-incubated with Rosu or with different stress signaling pathway inhibitors including mevalonate (100 μmol/L), acetyl-cysteine (NAC, 10 μmol/L), NADPH oxidase inhibitor (DPI, 10 μmol/L), and endothelial nitric oxide synthase inhibitor (LNMA, 1 mmol/L) before adding Hcy. Apoptosis rate was evaluated by fluorescence activated cell sorting (FACS) analysis. ROS levels were detected by 2’,7’-dichlorodihydrofluorescein diacetate (H2DCFH-DA). NADPH oxidases were evaluated with lucigenin-enhanced chemiluminescence. Expression of Nox4 mRNA was measured by RT-PCR. Results Rosu remarkably inhibited Hcy-induced ROS accumulation and apoptosis of EPCs, and antagonized Hcy-induced activation of NADPH oxidase and Nox4 mRNA expression. Nox4 siRNA transfected EPCs with a similar effect. Conclusion The protective effect of Rosu on EPCs possibly involves inhibition of Hcy-induced activation of Nox, ROS accumulation, and apoptosis of EPCs through Nox4 dependent mechanisms.