小鼠血清补体替代途径溶血活性测定的新方法  

A new method for measurement of the alternative pathway activity of mouse serum complement

作　　者：孙黔云[1] 叶巧玲[1] 闫银萍[1] 

机构地区：[1]贵州省中国科学院天然产物化学重点实验室,贵州贵阳550002

出　　处：《中国药理学通报》2011年第11期1619-1622,共4页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30560035);国家重点基础研究发展计划(973计划)资助项目(No2007CB516813);贵州省优秀青年科技人才资助项目(No黔科合人字2005-0510号)

摘　　要：目的针对现有补体溶血活性测定方法不适宜测定小鼠血清补体的缺点,构建一种血清用量少、灵敏度高的小鼠血清补体替代途径溶血活性测定的新方法。方法选用具有代表性的3个小鼠品系KM、BALB/c和C57BL/6小鼠的血清补体作为测定材料,以兔红细胞为溶血靶细胞,利用眼镜蛇毒因子(CVF)特异激活补体替代途径的特点,构建和优化小鼠血清补体替代途径溶血活性测定的方法,并采用该方法测定两种抗补体蛋白对小鼠体内血清补体活性的影响。结果以CVF作为激活剂成功构建了测定小鼠血清补体替代途径溶血活性的微量新方法。3个品系小鼠的血清在5～20μl的范围内即可分别达到合适的溶血度。该方法明显减少了血清用量,并能有效提高溶血度。结论基于CVF特异激活补体替代途径的特点而构建的小鼠血清补体替代途径溶血活性测定新方法,血清用量少、灵敏度高、稳定性好,适用于测定小鼠血清补体的溶血活性。Aim To constitute a highly sensitive method to measure the alternative pathway activity of mouse serum complement by using small amount of serum.Methods Three mouse strains KM,BALB/c,and C57BL/6 were chosen to be the source of mouse serum complement.Based on the fact that the alternative complement pathway could be specifically activated by cobra venom factor（CVF）,a simple assay was devised for determining the alternative pathway of mouse serum by using rabbit erythrocytes as target cells.The developed assay was then used to determine the changes of serum complement activity after two kinds of anticomplementary proteins were given to mice.Results The new sensitive assay for measuring the alternative pathway activity of mouse serum was successfully constituted in this research by employing CVF as activator.The sera of the three mouse strains in volume of 5~20 μl achieved suitable hemolysis.Conclusion The new assay for determining the alternative pathway of mouse serum by using CVF as activator possesses merits of high sensitivity,good stablity,and less serum consumption.This practical assay is very useful for measuring the alternative pathway activity of mouse serum complement.

关 键 词：小鼠血清 补体溶血活性 补体替代途径 补体激活 眼镜蛇毒因子 抗补体蛋白 

分 类 号：R976[医药卫生—药品] R392.5[医药卫生—药学]