水提取法和仿生提取法研究菲牛蛭不同炮制品的体外抗凝活性

目的:采取不同的提取方法研究菲牛蛭炮制前后抗凝活性变化。方法:分别采用水提取法及模拟胃肠道环境的仿生提取法提取菲牛蛭活体冻干品、清水吊干品、滑石粉烫制品、酒浸闷烘品中的活性成分,测定活化部分凝血酶时间(APTT)、凝血酶原时间(PT)、凝血酶时间(TT)、抗凝血酶活性4个凝血指标,并采用Lorry法测定其蛋白含量,综合评价2种提取方法下的菲牛蛭及其炮制品的抗凝活性。结果:无论是水提取法,还是仿生提取法,APTT、PT、TT均显示炮制后抗凝活性降低,抗凝活性顺序为活体冻干品>清水吊干品>酒浸闷烘品>滑石粉烫制品,此结果与抗凝血酶活性及蛋白含量顺序一致。结论:高温炮制方法会降低菲牛蛭的抗凝活性。

In Vitro Anticoagulant Activity of Different Processed Products of Poecilobdella manillensis by Water Extraction and Bionic Extraction

Objective:Different extraction methods were used to study the changes of anticoagulant activity before and after processing of Poecilobdella manillensis.Methods:bionic extraction method and water extraction were used respectively to extract the anticoagulant active components in Poecilobdella manillensis living lyophilized product,hanging dry products,talcum powder fried products and wine immersing-baked products.Activated partial thromboplastin time(APTT),prothrombin time(PT),thrombin time(TT),and antithrombin activity were selected as anticoagulant activity index to evaluate the anticoagulant activities of different processed Poecilobdella manillensis.The protein content in each extract of the processed products was determined by Lorry method.Results:Whether water extraction or biomimetic extraction,the results from APTT,PT and TT all showed that the anticoagulant activity decreases after processing.the anticoagulant activity sequence was as follows:living lyophilized product>hanging dry products>wine immersing-baked products>talcum powder fried products,which is consistent with the anti-thrombin activity and protein content of different processed products of the Poecilobdella manillensis.Conclusions:Therefore,the traditional processing method will reduce the anticoagulant activity of the Poecilobdella manillensis,and it is preferable to use the lyophilized product or the hanging dry products as the anticoagulant drug in clinical use.