目的研究丹参酮ⅡA对高血压病患者血清损伤后的血管内皮细胞(VEC)的影响,进一步探讨丹参酮ⅡA保护VEC的作用机制。方法利用15%的高血压病患者血清损伤正常培养的人脐静脉内皮细胞(ECV-304),用不同质量浓度的丹参酮ⅡA(10、20、40μg/mL)作用于损伤后的细胞24 h,运用四甲基偶氮唑蓝(MTT)法检测细胞的活性;单细胞凝胶电泳(SCGE)技术检测细胞DNA的损伤;逆转录-聚合酶链式反应(RT-PCR)技术检测内皮细胞血栓调节蛋白(TM)和血管性假血友病因子(vWF)mRNA的表达。结果与模型组比较,20μg/mL和40μg/mL的丹参酮ⅡA能增加内皮细胞的活性(P〈0.05);3个质量浓度的丹参酮ⅡA均能减轻内皮细胞DNA的损伤(P〈0.01);3个质量浓度的丹参酮ⅡA都可以下调TM mRNA表达(P〈0.01);20μg/mL和40μg/mL的丹参酮ⅡA可以下调vWF mRNA表达(P〈0.01)。结论丹参酮ⅡA对高血压病患者血清损伤后的ECV-304有保护作用,其作用机制可能与丹参酮ⅡA减轻细胞的DNA损伤有关。
AIM To study the effect of tanshinoneⅡA on the vascular endothelial cell(VEC) injury induced by the serum of the patients with hypertension,and to explore the mechanism of action further.METHODS The human umbilicus vein endothelial cells(ECV-304) were injured by the 15% serum of the patients with hypertension,then treated with different concentrations of tanshinoneⅡA(10 μg/mL,20 μg/mL,40 μg/mL) for 24 hours.And the activity of the VEC was determined by MTT assay.The DNA damage to VEC were determined by single cell gel electrophoresis(SCGE).The expression of the mRNA of thrombomodulin(TM) and von willebrand factor(vWF) were determined by the reverse transcriptase-Polymerase chain reaction(RT-PCR).RESULTS To compare with the model group,20 μg/mL and 40 μg/mL tanshinoneⅡA can increase the activity of the VEC(P〈0.05),all the three doses tanshinoneⅡA can reduce the cell DNA damage and decrease the expression of the mRNA of TM significantly(P〈0.01),and 20 μg/mL and 40 μg/mL tanshinoneⅡA can decrease the expression of the mRNA of vWF(P〈0.05).CONCLUSION TanshinoneⅡA has a protective effect on the VEC injured by the serum of patients with hypertension,and its possible mechanism reduces the damage to the cell DNA.