【作者】 彭志允；

【导师】 陈利国；

【作者基本信息】 暨南大学 ， 中西医结合临床， 2008， 硕士

【摘要】 目的:研究丹参酮ⅡA(TanⅡA)对高血压病血瘀证患者血清损伤后的血管内皮细胞(VEC)的影响,进一步探讨TanⅡA保护VEC的作用机制。方法:利用15%的高血压病血瘀证患者血清损伤正常培养的人脐静脉内皮细胞(ECV-304),用不同浓度的TanⅡA(10μg/ml、20μg/ml、40μg/ml)作用于损伤后的细胞24h,运用四甲基偶氮唑蓝(MTT)法检测细胞的活性;硫代巴比妥酸法检测细胞上清液中丙二醛(MDA)含量;黄嘌呤氧化酶法检测细胞上清液中超氧化物歧化酶(SOD)的活力;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TanⅡA能增加内皮细胞的活性(P＜0.05);20μg/ml和40μg/ml的TanⅡA可降低细胞培养液中MDA的含量(P＜0.05);40μg/ml的TanⅡA可增加SOD的活性(P＜0.01);三个浓度的TanⅡA都能减轻内皮细胞DNA的损伤(P＜0.01);三个浓度的TanⅡA都可以下调TMmRNA表达(P＜0.01);20μg/ml和40μg/ml的TanⅡA可以下调vWFmRNA表达(P＜0.01)。结论:TanⅡA对高血压病血瘀证患者血清损伤后的ECV-304有保护作用,其作用机制可能与TanⅡA抗氧化损伤,减轻细胞的DNA损伤有关。更多还原

【Abstract】 Object:To study the effect of TanⅡA on the vascular endothelial cell(VEC) injury induced by the serum of the patients of blood stasis syndrome(BSS) associated with hypertension disease,and discuss the mechanism of this effect further.Methods:The human umbilicus vein endothelial cells(ECV-304) was injured by the 15% serum of the patients of BSS associated with hypertension disease,then treated with different concentrations of TanⅡA(10μg/ml、20μg/ml、40μg/ml) for 24 hours.And the activity of the VEC was determined by MTT assay.The content of Malondialdehyde(MDA) in cell medium was measured by thlobarbiturie acid reaction and the activity of superoxide dismutase(SOD) measured by xanthosine oxidase method.The DNA damage of the VEC was determined by Single cell gel electrophoresis(SCGE).The expression of the mRNA of thrombomodulin(TM) and von willebrand factor(vWF) was determined by the reverse transcriptase-Polymerase chain reaction(RT-PCR).Results:Compare with the model group,the 20μg/ml and 40μg/ml TanⅡA can increase the activity of the VEC and decrease the MDA content in cell medium(P＜0.05),the 40μg/ml TanⅡA can increase the SOD activity in cell medium significantly(P＜0.01),all the three doses TanⅡA can reduce the cell DNA damage and decrease the expression of the mRNA of TM significantly(P＜0.01),and the 20μg/ml and 40μg/ml TanⅡA decreased the expression of the mRNA of vWF(P＜0.05).Conclusion:TanⅡA has a protective effect on the VEC injured by the serum of the patients of BSS associated with hypertension disease,and its possible mechanisms are antioxidant and reduce the damage of the cell DNA.更多还原