【作者】 宋文龄；

【导师】 孙景辉；

【作者基本信息】 吉林大学 ， 内科学， 2007， 博士

【摘要】 血栓性疾病是一种常见的心脑血管病,常表现为心肌梗死、缺血性脑梗死、静脉血栓栓塞。每年每千人中有1-3人发生不同形式的血栓性疾病,严重危害着人类的生命和健康。抗血栓药物的使用是血栓疾病防治的重要环节,现有的药物由于存在各种各样的问题,有的疗效不好,有的价格昂贵,有的不良反映较多,较严重。显然,寻找安全、价廉及有效的抗血栓药物极为有意义。蚓激酶具有溶解血栓,抑制血小板聚集,降低纤维蛋白原及改善血栓形成前高凝状态的作用已被实验所证实,因而认为其具有抗血栓形成的活性。本研究通过基因工程手段建立高效表达的蚓激酶转基因山羊乳腺生物反应器,并对表达的基因工程蚓激酶首次进行急性毒性试验和抗血栓作用的体内生物活性检测。本研究实现了由羊乳中高效表达蚓激酶,并证实基因工程蚓激酶无明显的急性毒性作用,有明显的抗血栓作用,为直接饮用羊乳治疗血栓性疾病奠定基础。更多还原

【Abstract】 The purpose of the work is to establish an efficiency express Goat Mammary gland bioreactor of lumbrokinase by genetic engineering way, and lumbrokinase can be expressed in milk, finally to check its function. To drink milk and treat thrombus with the purpose of health. Now, three works have been done: lumbrokinase’s espression in mammal cells; The acute toxic effects of genetic engineering lumbrokinase; to check antithrombus function of genetic engineering lumbrokinase.To constract pbLK and pb-LK-tpa specialized vector expresses in mammary gland. Then the two expression vectors were injected in goat mammary gland. Fibin agarose plate assay indicated both of the two plasmides expressed in milk. The expression quantity of pIbCP-LK-tpa was higher than pIbCP-LK. It was the first time that lumbrokinase expressed in goat mammary gland and it provided a dependable guarantee for the establishment of transgenic goat and the expression of lumbrokinase in mammary gland.The acute toxic effects of genetic engineering lumbrokinase were evaluated .The routine acute toxic test on mice was performed by perfusing the mice with genetic engineering lumbrokinase. Results: The maximum tolerance dosage of mice in genetic engineering lumbrokinase through mouth was 4.002W IU/kg BW; no microscopic changes were found in the heart, liver, spleen, lung and kidney. It is proved that genetic engineering lumbrokinase had no acute atoxicity.Effect of genetic engineering lumbrokinase on Thrombus Formation in the Arteries in Rats. Methods to observe the thrombosis and fibrinolysis effect of OT in the arteries through electric stimulation of common carotid artery. In vivo and in vitro thrombosis, and in vivo fibrinolysis tests were used to evaluate the effects of the drug on the fibrinolysis system. To observe anti- thrombus function were examined in the normal saline group and high-dose, middle-dose, low-dose groups of genetic engineering lumbrokinase. Results High-dose, middle-dose, and low-dose groups of genetic engineering lumbrokinase could prolong OT and decrease Thrombus Formation. it reduced thrombus weight and length significantly; it also showed a strong enhancement on fibrinolysis. Conclusion genetic engineering lumbrokinase could significantly inhibit the formation of thrombus in the arteries, and also increase fibrinolysis.更多还原