临床实用的新思路和新方法:体外血液生命系统药物治疗个体化试验  

Clinical Utility of New Ideas and New Ways:In-Vitro Individual Drug Treatment Test in Blood Living System

作　　者：郭峰[1] 

机构地区：[1]第二军医大学长海医院输血科血液免疫学研究室,上海200433

出　　处：《临床医学工程》2011年第1期4-6,共3页Clinical Medicine & Engineering

基　　金：国家自然科学基金项目"现代系统免疫学自然与分离体系实验研究"(批准号30671940)

摘　　要：目的建立体外静脉注射药品测试系统,以确定药物注射后是否引发血液中快速先天免疫炎症反应,保证避免可以致死的天然免疫炎症反应的发生。方法采用体外血液生命系统药物治疗个体化试验,设计为三组:新药实验组:0.2毫升新药加到0.6枸橼酸抗凝的新鲜正常人全血中;阳性对照组:0.2毫升(热灭活)大肠杆菌悬液(浓度为6×109),加到0.6毫升枸橼酸抗凝的新鲜新鲜正常人全血中;阴性对照组:0.2毫升生理盐水加入新鲜的0.6毫升枸橼酸抗凝全血中。充分混匀,37℃孵育1小时,2000转/分离心5分钟;测定与统计各组IL-8/IL-10和IL-8/DARC指标,按上述公式,求出新药实验组和阳性对照组的激活率(药物的影响率)。结果根据新药实验组和阳性对照组的激活率的比较,可判定新药是否具有免疫原活性。结论此测试系统操作简单,只需清洁操作,不需无菌操作,也不需加人工培养液,但注意点是要新鲜血,采血后应尽快做实验,而且必须是枸橼酸抗凝,不能灭活补体系统,也就是说不能用EDTA抗凝,否则血液失去生命信息完整的网络关系。只要认真按以上要求,细心认真操作,一定能成功。此创新方法对疾病治疗研究有帮助。Objective To establish an in-vitro intravenous drug test system to determine blood innate immune inflammatory responses caused by the drugs.This test can determine whether intravenous drugs can cause rapid innate immune and inflammatory responses.Methods Design for the 3 groups：new drugs experimental group：0.2 ml new drugs added to 0.6 ml of fresh citrate anticoagulant normal human whole blood;positive control group：0.2 ml（heat inactivated） E.coli（concentration of 6 × 109） suspension were added to 0.6 ml of citrate anticoagulant fresh normal human whole blood;negative control group：0.2 ml physiological saline were added to 0.6 ml of citrate anticoagulant fresh normal human whole blood In the fresh blood culture system,37 ℃ 1 hour incubation,then were centrifuged 2 000 rounds per minute for 5 minutes.In the fresh blood culture system,expression（mean fluorescence intensity） of DARC on red blood cells（FY6：IL-8 receptor） and IL-8/IL-10 and IL-8 /DARC and the activation rate（drug effect rate） were observed.Results The activation rates of new drug test group and positive group compared to determine whether immune activity of drugs.Conclusion This test system is simple,just a clean operation,without aseptic operation,nor do they need additional artificial medium,but the point is to note that the new blood,blood as soon as possible the experiment,and must be citrate anticoagulation that can not inactivate the complement system,that can not be used EDTA anticoagulant（blood loss of life or integrity of the information network of relationships）.As long as the above requirements are met,the experiment would be successful.This innovative approach is conducive to disease therapy research.

关 键 词：体外药物治疗个体化试验模型 血液天然免疫炎症反应 IL-8/IL-10比值 IL-8/DARC比值 药物激活率 

分 类 号：R392-33[医药卫生—免疫学] R446.11[医药卫生—基础医学]