Regulator Of Endogenious PAI-1 Expression From Chinese Herb And Its Pharmacology

The plasminogen activator inhibitor type 1 (PAI-1) is a primary factor in fibrinolysis system. This kind of therapeutic medicine using PAI-1 as the target, which is among the hotspots of thrombolytic researching. Up to now, there are some disadvantages in their application, such as short-lived half life, narrow-ranged safe dosage, expensive price etc. In this thesis, drug screening was started with Chinese Herb and PAI-1 was used as a target. The purpose is to find PAI-1 down-regulating Chinese Herb. This is very helpful for avoiding the above shortages and for the development of new anti-thrombosis agents.A realtime RT-PCR was constructed for detecting PAI-1 mRNA expression from Human microvascular endothelial cell (HMEC-1). SYBR Green I was used in development of real-time PCR amplification standards curves. The stability, specificity and sensitivity of the method were evaluated. With the high specificity the method could be used to detected as low as 103 copies PAI-1mRNA with the linear range from 103 to 1010 copies. The standard curves showed high correlations(R2 > 0.990). The coefficient of variation values for both intra-assay and inter-assay ranged from1.56%～4.79% to 8.54%～10.23%.Two Chinese herbs were found have the activity to forbid PAI-1 expression. They were chuanxiong and danshen. Compared with negative control, the drug chuanxiong could down-regulated PAI-1 mRNA 1.11 folds lower.Chuan Xiong were extracted by different solvents. The concentrations of tissue-type plasminogen activator(t-PA) and PAI-1 in HMEC-1 were assayed using the enzyme linked immunosorbent assay technique (ELISA).Also,the activities of t-PA and PAI-1 were assayed using Chromogenic assay technique(CSA), mRNA of t-PA and PAI-1 were assayed using realtime RT-PCR. The concentrations of t-PA in ethanol extraction were higher than control group.