上海地区人群抗凝蛋白C基因突变和多态性研究

目的研究上海地区人群抗凝蛋白C(PC)活性的正常范围和PC基因突变及多态性的分布;研究活化蛋白C抵抗(APC-R)的分布及其与相应已知基因突变的关系。方法采用发色底物法检测867例上海健康人PC活性,采用酶联免疫吸附试验(ELISA)法检测PC抗原,采用凝血法检测APC-R,采用聚合酶链反应(PCR)扩增PC基因所有外显子及其侧翼序列和5′、3′非翻译区,采用直接测序法检测所有扩增片段。采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)法检测APC-R相关多态性。结果 PC活性和APC-R的正常范围分别为67%～147%和114.7～195.2S。不同性别研究对象PC和APC-R测定结果间差异有统计学意义(P<0.05)。PC活性随年龄增长而增加(P<0.01)。在对10例PC活性降低标本的基因检测中发现2个基因突变,即Ala121Thr(G3406A)和Pro247Ser(C8454T)。据此,计算出上海地区健康人群PC基因突变的频率为0.23%。在对15例APC-R阳性标本的检测中未发现FV多态性位点(FV Arg306Thr、Arg306Gly、Arg506Gln、His1299Arg和Ile359Thr)和PT G20210A。结论 PC基因突变的频率为0.23%;APC-R与已知的FV多态性位点无关;PC基因突变Ala121Thr和Pro247Ser可能是中国人易栓症的危险因素。

Genetic Mutations and Polymorphisms of Protein C in General Shanghai Population

Objective To study the normal range of protein C activity, and the underlying gene mutations in the normal Shanghai population of China. To detect the relationship of APC-R and mutations reported correspond to it in normal Chinese populaion. Methods PC activity were assayed by using chromogenic substrate method and APC-R were assayed by a clotting method. PC gene of all exons and the 5′, 3′-untranslated regions were amplified by PCR. The SNPs related to APC-R were analyzed by PCR restriction fragment length polymorphism （RFLP）. Results The normal range of PC activity and APC-R were 67%-147% and 114. 7-195. 2 S. The difference between men and women in PC activity and APC-R were statistical significance （P〈0. 05）. PC activity grew with age. We detected two mutaions, Ala121Thr （G3406A） and Pro247Ser （C8454T）, in those 10 persons whose PC activity were under normal. We did not detect any -positive results in the RFLP tests related to APC-R. Conclusion The incidence of PC gene mutation is 0. 230/00. There is no relationship between APC-R and SNPs reported correspond to it. PC mutations, Ala121Thr and Pro247Ser, may be the risk factor of thromboembolism in Chinese people.