基于体外抗凝血实验研究铅污染对水蛭抗凝血作用的影响

Effect of lead pollution on anticoagulant effect of Whitmania pigra based on in vitro anticoagulation experiment

水蛭具有良好的抗凝血活性,是众多心脑血管成药的原料药之一。该研究基于体外抗凝血实验(APTT法和PT法)探究铅污染对水蛭抗凝血作用的影响,选取目前市场上流通最广的水蛭品种--蚂蟥Whitmania pigra,对其进行为期50 d的不同铅污染生长环境下的模拟养殖,按药典方法加工后获得水蛭药材。利用全自动凝血仪测定加入不同处理组水蛭提取液的血浆的活化部分凝血酶时间(APTT)和凝血酶原时间(PT),通过比较各组APTT及PT的差异,研究铅污染对水蛭抗凝血作用的影响。结果表明水蛭药材中的铅残留量随着养殖土壤中铅含量的增加而增加,铅高残留水蛭组(Pb-H)的铅残留量为(10. 66±2. 79) mg·kg^-1,超过了2015年版《中国药典》规定的铅限量。体外抗凝血实验结果显示:与生理盐水组相比,水蛭提取液能显著延长APTT;铅高残留水蛭组的APTT较空白水蛭组的短,且存在显著性差异。不同处理组水蛭提取液对PT的影响不大。结果表明生长环境污染是造成水蛭药材铅残留量超标的重要因素之一,铅污染会降低水蛭的抗凝血作用,影响其质量和临床疗效。

Leech has a good anticoagulant activity and is one of the raw materials for treatment of many cardiovascular and cerebrovascular diseases. This study was based on in vitro anticoagulant experiments( APTT and PT) to investigate the effects of lead contamination on the anticoagulant effect of leech. At present,the Hirudo circulating in the market are dominated by Whitmania pigra,therefore Wh. pigra were cultivated under a different lead pollution for 50 days. Then,the effects of Wh. pigra extract,extracting from different cultivating environment,on activated partial thrombin time( APTT) and prothrombin time( PT) were determined by automatic coagulation instrument. The results showed that the Wh. pigra extract significantly prolonged the APTT compared with the saline group.The APTT of the lead-high residual Wh. pigra was shorter than that of the blank Wh. pigra. The Wh. pigra extracts from different treatment groups had little effect on PT. The results showed that the lead residue in the Wh. pigra increased with the increase of lead in the cultured soil,the lead residual of the Pb-H group was( 10. 66±2. 79) mg·kg^-1,which exceeded the lead limit specified in the 2015 edition of the Chinese Pharmacopoeia. The results indicated that growth environment pollution is one of the important factors causing excessive lead in Wh. pigra. Lead pollution will reduce the anticoagulant effect of Wh. pigra and affect its clinical efficacy.