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目的比较湿法超微粉碎及水煎煮提取法对鼠妇溶栓抗凝活性蛋白的影响,并探究其溶栓抗凝活性的机理。方法比较两种不同提取法所得鼠妇提取物得率及蛋白含量的差异,并采用硫酸铵分级盐析初步纯化鼠妇活性蛋白,纤维蛋白平板法检测鼠妇活性蛋白对纤维蛋白的降解作用及溶栓机理,全自动凝血因子分析仪测定鼠妇活性蛋白对凝血酶时间(TT)、凝血酶原时间(PT)、活化部分凝血活酶时间(APTT)的影响,考察温度对鼠妇活性蛋白的影响。结果两种不同提取方法在提取物得率、蛋白含量存在较大差异;其中,鼠妇的水煎煮提取物未检测出溶栓活性,而湿法超微粉碎提取的蛋白具有很强的溶栓活性,其活性蛋白主要集中在60%~80%硫酸铵饱和度范围内,通过激活纤溶酶原的方式降解纤维蛋白,并且能明显延迟TT、APTT,但对于PT无明显作用;而水煎煮提取物对TT、PT、APTT均无延迟作用。结论湿法超微粉碎提取法能很好地保留鼠妇溶栓抗凝活性蛋白,而水煎煮提取的鼠妇蛋白无溶栓抗凝活性。
Objective To compare the effect of wet ultrafine pulverization and decoction method on thrombolytic and anticoagulant proteins in rats and explore the mechanism of thrombolytic and anticoagulant activity. Methods To compare the differences in the yield and protein content of extracts from rat extracts obtained by two different extraction methods. Purification of rat active protein by ammonium sulfate fractional salting-out and the degradation of fibrin by fibrin plate assay Thromboplastin time, thromboplastin time (PT) and activated partial thromboplastin time (APTT). The effect of temperature on the expression of murine active protein Impact. Results Two different extraction methods showed great differences in the yield and protein content of the extract. Among them, the thrombolytic activity was not detected in the decoction of the rat woman, and the protein extracted by the wet ultrafine pulverization had a strong solubility Suppository activity. The active proteins mainly concentrated in the range of 60% -80% ammonium sulfate saturation, degrading fibrin by activating plasminogen, and delayed TT and APTT obviously, but had no obvious effect on PT. However, water Decoction extract on TT, PT, APTT no delay. Conclusion The wet ultrafine pulverization extraction method can well preserve the thrombolytic and anticoagulant active protein in rats, while the thrombolytic and anticoagulant anticoagulant activity of the rat extract extracted by water decoction.