湿法超微粉碎和水煎煮提取法对鼠妇蛋白溶栓抗凝活性的影响及机制初探  

Comparison of Thrombolytic and Anticoagulant Activities of Proteins of Porcellio scaber Latreille Extracted by Wet Superfine Grinding Technology and Water Decoction and Exploration of Their Mechanisms

作　　者：田周[1,2] 李博[1,2,3] 郭立玮[1,2] 吴勉华[3] 程海波[3] 朱华旭[1] 

机构地区：[1]南京中医药大学江苏省中药资源产业化过程协同创新中心,江苏南京210023 [2]南京中医药大学中药复方分离工程重点实验室,江苏南京210029 [3]南京中医药大学第一临床医学院,江苏南京210046

出　　处：《中药新药与临床药理》2015年第2期169-174,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金(81274096;81303230);江苏省科技厅产学研联合创新资金-前瞻性研究项目(BY2012036);江苏省科技支撑计划-社会发展项目(BE-2012763);中国博士后科学基金(2013M541704);江苏省博士后科学基金(1301133C)

摘　　要：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 active thrombolytic and anticoagulant proteins of Porcellio scaber Latreille extracted by wet superfine grinding technology and water decoction,and to study their thrombolytic and anticoagulant mechanisms.Methods We compared the difference of the yield of extract and the protein content extracted by wet superfine grinding technology and water decoction.Fractional precipitation method with ammonium sulfate was applied to purify the active proteins.The fibrinolysis activity and the mode of fibrinolyctia action of the active proteins were measured by agarose-fibrin plate.Moreover,prothrombin time（PT）,thrombin time（TT）,activated partial thromboplastin time（APTT）were measured by the automatic blood agglutination instrument.The effect of temperature on the obtained active proteins was also investigated.Results There were great differences in the yield of extract and the protein content extracted by water decoction and ultra-fine pulverization in wet processing.The extract obtained by water decoction had no thrombolytic activity,but the extract obtained by ultra-fine pulverization in wet processing could stimulate the secretion of plasminogen activator and dissolve fibrin.Besides,at the range of saturation from 60% to 80 %,the active proteins extract obtained by wet ultra-fine pulverization had the higher thrombolytic activity;they could also delay APTT and TT,but had no effect on PT.However,the protein extracted by water decoction had no effect on TT,PT,and APTT.Conclusion The wet superfine grinding technology can maintain maximally the active components of Porcellio scaber Latreille,and the active proteins have obvious thrombolytic activity and anticoagulant function.However,the proteins obtained by water decoction have no thrombolytic and anticoagulant activities.

关 键 词：鼠妇 活性蛋白 湿法超微粉碎 溶栓 抗凝血 

分 类 号：R285.5[医药卫生—中药学]