活化血小板释放产物对内皮细胞的损伤及丹酚酸B的保护效应

目的探讨活化血小板释放产物(PLTaRP)对内皮细胞的损伤作用及丹酚酸B的保护效应。方法分离健康人外周血血小板,以二磷酸腺苷激活后提取PLTaRP,并与EA.hy926人脐静脉内皮细胞株共培养,分为对照组、模型组、阳性对照组、低剂量组、中剂量组、高剂量组,检测各组细胞活力及乳酸脱氢酶(LDH)释放水平。DAPI染色观察细胞核形态,比较各组细胞凋亡率。结果与对照组比较,模型组12 h内LDH含量持续升高,并呈时间依赖趋势(P<0.01),模型组、阳性对照组、低剂量组和中剂量组细胞存活率明显减低(P<0.05,P<0.01),高剂量组细胞存活率差异无统计学意义(P>0.05)。与模型组比较,低、中、高剂量组LDH含量明显降低(P<0.01)。结论 PLTaRP可刺激EA.hy926细胞株LDH释放增加,并呈时间依赖趋势,刺激12 h可使细胞活力明显下降。丹酚酸B可减少PLTaRP造成的LDH释放。高剂量丹酚酸B可明显抑制细胞凋亡。

Endothelial cell damage induced by activated platelet released products and the protective effect of salvianolic acid B

Objective To observe the endothelial cell damage induced by activated platelet-released products(PLTaRP) and the protective effect of salvianolic acid B(SalB).Methods PLTaRP was extracted from peripheral platelets in healthy people activated by ADP and was incubated with the human umbilical vein endothelial cell of cell strain EA.Hy926.The LDH release level in culture supernatants and cell viability were detected in the control group,model group,positive control group and three doses SalB preconditioning group(1,5,10μg/ml SalB preconditioning 24 h, PLTaRP stimulating 12 h).DAPI staining was employed to observe morphological changes of cell nuclus,and the apoptotic cell rates in each group were compared.Results PLTaRP could lead to gradual increase of LDH level in cell culture supernatants(compared with control group,P< 0.01) within 12 h.LDH release level in model group was higher than the control group(P< 0.01),and SalB preconditioning could reduce it significantly in a dose-dependent manner(P< 0.01).The cell viabilities in model,low and medium dose group were significantly lower than those in the control group(P<0.05,P<0.01).The typical apoptotic morphological changes were observed in model group,and high dose SalB preconditioning could reduce apoptotic cell rates sig- nificantly(P<0.01).Conclusions PLTaRP could stimulate EA.Hy926 LDH release was increased in a time-dependent manner,and cell viability was declined significantly at endpoint.SalB preconditioning may reduce the LDH level induced by PLTaRP,and has a protective effect on cell survival.High dose SalB preconditioning could inhibit cell apoptosis caused by PLTaRP.