人附红细胞体病发病机制及蒙药治疗的实验研究

Abstract: Human Eperythrozoonosis is a newly discovered kind of zoonosis that has a part distribution in our Autonomous Region, infection rate up to fifty-five percent in pastoral areas. It harms to our health and influences the development of stock rising seriously. At present，because there is so little known about the underlying defect in Eperythrozoonosis and its pathogenesis , no specific treatment is available, and it has a highest relapsed rate after treatment. Our study Protocol as follows:.1. The models of Human Eperythrozoonosis in rabbits will be made. Eperythrozoon attacks red blood cells (RBC) mainly so that erythrocyte membrane appears pathological changes. C3b receptors of RBC、erythrocyte membrane surface antigen and ATP enzyme will be measured, at the same time, IL-1、TNF、CRP、hematological and immunological check be examed，and so an. Through these we can explore the mechanisms of fever、anemia and immune function changes. Pathological changes of erythrocyte membrane and invasiveness pathway、proliferation way of Eperythrozoon will be observed by electronic microscopy..2. Through making models of human Eperythrozoon in vitro culture，we can carry on drug screening and compare curative effects..3. In Mongolian drugs intervention of Human Eperythrozoonosis models in rats, we will screen new efficient drugs. By the research of pathogenesis that these results would provide the theory basis and scientific support for clinical diagnosis、treatment and prevention..This research project brings forth new ideas about technique for human Eperythrozoon in vitro culture, meanwhile, new treating and efficient drugs of Mongolian drugs will be screened.

人附红体病是一种新发现的人畜共患病，在牧区人群感染率高达55%以上，我区部分地区已有流行，严重危害人的健康，给畜牧业发展带来严重影响。目前对此病尚无特效药物，治疗后极易复发，对其发病机理尚不清楚。本项目研究方案：1、制作兔人附红体感染模型，以附红体主要侵袭红细胞和红细胞膜的病理改变为重点，通过测定红细胞C3b受体、红细胞膜表面抗原CD47分子、红细胞膜ATP酶活性，同时测定IL-1、TNF和CRP、血液学及免疫学等指标，来探讨发热、贫血及免疫功能的变化；通过电镜观察研究附红体的侵袭力和增值方式及细胞膜的受损程度。2、建立人附红体体外培养，并进行药物筛选，比较药物疗效。3、用蒙药干预感染人附红体大鼠模型，从蒙药中筛选新的治疗药物和特效药物。通过发病机理的研究，为临床诊断、治疗和预防提供理论基础与科学依据。本项目创新之处在于建立人附红细胞体体外培养技术，在蒙药中筛选新的治疗药物及寻找特效药物。

通过对人附红细胞体重度感染患者的血液学检测发现，人附红细胞体重度感染者可出现溶血性贫血：既有血管内溶血、血管外溶血，也有自身免疫性溶血，其发生机制与机体红细胞免疫功能受损有关；通过筛选，初步建立人附红细胞体体外培养体系：将制备好的阳性红细胞泥加入添加了40%胎牛血清的RPMI-1640培养液中，于37℃、5%CO2条件下培养，按原代培养条件传代培养，可连续传至15代。并在此基础上选用5种药物进行体外药敏实验，人附红细胞体对药物的敏感程度依次为青蒿琥酯、左氧氟沙星、多西环素、林可霉素、庆大霉素；另外，项目组还成功复制出人附红体感染的大鼠模型：SPF级SD大鼠腹腔注射人附红体3天后的感染率与对照组相比显著上升。通过对感染人附红体的SD大鼠的研究发现：附红体可能亦在骨髓中增殖；被感染的大鼠体液免疫、细胞免疫及红细胞的免疫功能均发生改变。本课题通过临床试验、体外培养及动物实验，探讨了人附红体病发病时红细胞及机体免疫功能的变化，初步揭示了人附红细胞体病的发病机制，初步筛选出敏感性治疗药物，为今后进一步研究其发病机制、开发新的治疗药物奠定了理论基础。