CAJ | 학술논문

目的观察川芎嗪(TMP)对凝血酶诱导脐静脉血管内皮细胞株ECV304表达组织因子(TF)的影响.方法 ECV304细胞培养采用RPMI 1640完全培养基;一期凝同法测总的细胞促凝活性(PCA);RT-PCR的方法检测TFmRNA.结果凝血酶增强ECV304细胞的促凝活性(PCA),并具有明显的量-效依赖关系(r=0.9602,P<0.01).通过乏FVII血浆与TF单克隆抗体法确认PCA来自TF活性.单用TMP(125～1000μg/ml)对ECV304细胞TF表达没有影响(P>0.05).在给予凝血酶之前30 min用TMP(125～1000 μg/ml)预处理ECV304细胞,TMP可呈剂量依赖式地抑制凝血酶(20 U/ml)诱导ECV304细胞PCA(r=-0.9644,P<0.01)和TF mRNA(r=-0.9576,P<0.05)增加的作用,在1000μg/ml时抑制作用达到最强;在凝血酶刺激ECV304细胞后4、6、8、10、12 h,TMP(1000 μg/ml)均可以抑制凝血酶诱导的ECV304细胞PCA增加,在8 h时最明显(P＜0.05).结论 TMP可抑制凝血酶诱导ECV304细胞TF的表达,这一作用是在mRNA水平上实现的.
목적 관찰천궁진(TMP)대응혈매유도제정맥혈관내피세포주ECV304표체조직인자(TF)적영향.방법 ECV304세포배양채용RPMI 1640완전배양기;일기응동법측총적세포촉응활성(PCA);RT-PCR적방법검측TFmRNA.결과 응혈매증강ECV304세포적촉응활성(PCA),병구유명현적량-효의뢰관계(r=0.9602,P<0.01).통과핍FVII혈장여TF단극륭항체법학인PCA래자TF활성.단용TMP(125～1000μg/ml)대ECV304세포TF표체몰유영향(P>0.05).재급여응혈매지전30 min용TMP(125～1000 μg/ml)예처리ECV304세포,TMP가정제량의뢰식지억제응혈매(20 U/ml)유도ECV304세포PCA(r=-0.9644,P<0.01)화TF mRNA(r=-0.9576,P<0.05)증가적작용,재1000μg/ml시억제작용체도최강;재응혈매자격ECV304세포후4、6、8、10、12 h,TMP(1000 μg/ml)균가이억제응혈매유도적ECV304세포PCA증가,재8 h시최명현(P＜0.05).결론 TMP가억제응혈매유도ECV304세포TF적표체,저일작용시재mRNA수평상실현적.
Objective To observe the effects of tetramethylpyrazine (TMP) on tissue factor (TF) expression induced by thrombin in human umbilical vein endothelium derived cell line ECV304. Methods The changes in the total cellular procoagulant activity (PCA) of ECV304 cells exposed to thrombin were observed with one-stage clotting assay. TF mRNA expression in the exposed cells was examined using semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). Results ECV304 cells stimulated with increasing concentrations of thrombin (1.25-20 U/ml) showed a gradual increase of PCA (r=0.9602, P<0.01). The application of FVII-deficient plasma and the monoclonal antibody of TF confirmed that the PCA of the cells mediated by TF activity. TMP at 125-1000 μg/ml alone did not affect TF expression in ECV304 cells (P0>0.20), TMP administered 30 min prior to thrombin exposure showed a significant concentration-dependent inhibitory effect on the increments of PCA (r=-0.9644, P<0.01) and TF mRNA expression (r=-0.9576, P<0.05) in ECV304 cells, and 1000 μg/ml TMP produced the strongest effect. In ECV304 cells stimulated with thrombin for 4, 6, 8, 10 and 12 h, TMP administration significantly inhibited the thrombin-induced PCA, and the effect was especially obvious at 8 h following thrombin exposure (P<0.05). Conclusion Thrombin induces TF expression in vascular endothelial cells, and this effect can be inhibited by TMP at the mRNA level.