Pawlowska Z, Pluskota E, Chabielska E, Buczko W, Okruszek A, Stec W J, Cierniewski C S
Department of Biophysics, Medical University of Lodz, Poland.
Thromb Haemost. 1998 Feb;79(2):348-53.
The effect of systemic inhibition of PAI-1 expression in rats by PS-16R, a phosphorothioate analogue of hexadecadeoxyribonucleotide complementary to a signal peptide coding sequence of rat PAI-1 mRNA, on PAI-1 activity in blood plasma and thrombus formation was studied in rat models for experimental thrombosis. In previous in vitro studies, oligonucleotides of PS-16R family have been shown to inhibit efficiently PAI-1 synthesis in endothelial cells by antisense mechanism. When PS-16R was administered intravenously as a single bolus injection (1 to 5 mg per rat), it produced a significant reduction in PAI-1 activity of blood plasma. This effect was both time- and concentration-dependent. Under the same conditions, three groups of rats were treated with control oligodeoxynucleotides such as PS-16R with double mismatches, with scrambled sequence, and an oligodeoxynucleotide with sense sequence (complementary to PS-16R), respectively. Based on these preliminary experiments, a low dose of 1.5 mg per rat was selected to produce approximately 20-30% reduction of PAI-1 activity in blood plasma and the effect of such a decrease in PAI-1 expression was tested on thrombus formation in two rat models for experimentally induced thrombosis. Such a limited decrease in PAI-1 activity produced a significant antithrombotic effect in the arterial thrombosis model. There was a profound delay in the occlusion time in rats treated with PS-16R when compared to control animals (80 +/- 3 and 55 +/- 3 h, respectively), although blood plasma activity of PAI-1 in the same groups of rats differed only by 20%. There was also a tendency to reduce both an incidence of venous thrombosis (58.33 and 68.11%, respectively) and thrombus weight (2.1 +/- 0.4 and 2.9 +/- 0.9 mg, respectively) in the animals treated with PS-16R. However, this effect was not significant. Thus, low dose of PS-16R through inhibition of PAI-1 synthesis in targeted cells in rats reduced PAI-1 activity in blood plasma and protected against arterial thrombus formation in the rat.
PS - 16R是一种与大鼠纤溶酶原激活物抑制剂-1（PAI - 1）mRNA信号肽编码序列互补的十六脱氧核糖核苷酸的硫代磷酸酯类似物，在实验性血栓形成的大鼠模型中研究了其对大鼠PAI - 1表达的全身抑制作用对血浆PAI - 1活性和血栓形成的影响。在先前的体外研究中，PS - 16R家族的寡核苷酸已被证明可通过反义机制有效抑制内皮细胞中PAI - 1的合成。当以单次推注静脉注射PS - 16R（每只大鼠1至5毫克）时，它可使血浆PAI - 1活性显著降低。这种作用具有时间和浓度依赖性。在相同条件下，三组大鼠分别用具有两个错配的对照寡脱氧核苷酸如PS - 16R、具有随机序列的寡脱氧核苷酸和具有正义序列（与PS - 16R互补）的寡脱氧核苷酸进行处理。基于这些初步实验，选择每只大鼠1.5毫克的低剂量以使血浆PAI - 1活性降低约20 - 30%，并在两种实验性诱导血栓形成的大鼠模型中测试这种PAI - 1表达降低对血栓形成的影响。PAI - 1活性的这种有限降低在动脉血栓形成模型中产生了显著的抗血栓作用。与对照动物相比，用PS - 16R处理的大鼠闭塞时间有显著延迟（分别为80±3小时和55±3小时），尽管同一组大鼠的血浆PAI - 1活性仅相差20%。在用PS - 16R处理的动物中，静脉血栓形成的发生率（分别为58.33%和68.11%）和血栓重量（分别为2.1±0.4毫克和2.9±0.9毫克）也有降低的趋势。然而这种作用并不显著。因此，低剂量的PS - 16R通过抑制大鼠靶细胞中PAI - 1的合成降低了血浆PAI - 1活性，并预防了大鼠动脉血栓的形成。
