血小板膜糖蛋白及其Ⅰ bα kozak位点基因多态性与缺血性脑血管病关系的研究

Study on association between platelet membrane glycoprotein and itsⅠbαkozak site gene polymorphism with ischemic cerebrovascular disease

目的探讨血小板膜糖蛋白(GP)Ⅰbα基因kozak位点基因多态性及GPⅠb/ⅠⅩ/Ⅴ复合物表达量在缺血性脑血管病患者的变化特点,明确二者在缺血性脑血管病发病机制中的作用。方法采用ELISA法测定GPⅠb/ⅠⅩ/Ⅴ的表达量。采用实时荧光聚合酶链反应(PCR)技术确定缺血性脑血管病组和对照组GPⅠbαkozak位点基因多态性。采用二磷酸腺苷诱导比浊法测定血小板聚集率。结果 (1)缺血性脑血管病组血清GPⅠb/ⅠⅩ/Ⅴ复合物的表达量与对照组比较,差异有统计学意义(P<0.05)。(2)缺血性脑血管病组和对照组比较,GPⅠbαkozak位点基因型分布和C等位基因频率比较,差异有统计学意义(P<0.05)。(3)缺血性脑血管病组和对照组在GPⅠbαkozak位点,C等位基因携带者血清GPⅠb/ⅠⅩ/Ⅴ复合物表达水平高于TT型,差异有统计学意义(P<0.05)。(4)血小板聚集率在缺血性脑血管病组明显高于对照组(P<0.01)。相关性分析结果显示,缺血性脑血管病组血小板聚集率与GPⅠb/ⅠⅩ/Ⅴ复合物表达水平呈正相关(r=0.684)。结论 (1)缺血性脑血管病患者血清高表达GPⅠb/ⅠⅩ/Ⅴ复合物。(2)缺血性脑血管病发病的易感基因是GPⅠbαkozak位点C等位基因。(3)血小板聚集率增高与缺血性脑血管病的发病有很大相关性。(4)GPⅠbαkozak位点基因突变作为遗传因素引起血清GPⅠb/ⅠⅩ/Ⅴ复合物表达增高。(5)GPⅠb/ⅠⅩ/Ⅴ复合物高表达导致了血小板聚集率的增高。

Objective To investigate the gene polymorphism of platelet membrane glycoprotein Ⅰbαgene kozak site and the change characteristics of expression amount of GPⅠb/ⅠⅩ/Ⅴcomplex in the patients with ischemic cerebrovascular disease and to determine their role in the pathogensis of ischemic cerebrovascular disease.Methods The enzyme-linked immunosorbent assay（ELISA）was adopted to measure the GPⅠb/ⅠⅩ/Ⅴ complex expression amount.The Taqman real-time PCR technology was used to determine the gene polymorphism of GPⅠbαkozak site in the ischemic cerebrovascular disease group and the healthy control group.The platelet agglutination rate was detected by using the ADP inducing turbidimetry.Results（1）The expression amount of serum GPⅠb/ⅠⅩ/Ⅴ complex had statistical difference between the ischemic cerebrovascular disease group and the control group（P〈0.05）.（2）The distribution of GPⅠbαkozak site genotypes and the C allele frequency had statistical difference between the ischemic cerebrovascular disease group and the control group（P〈0.05）.（3）The expression level of serum GPⅠb/ⅠⅩ/Ⅴ complex at the GPⅠbαkozak site in the carriers with C allele in the ischemic cerebrovascular disease group and the control group was higher than that with TT genotype,the difference was statistically significant（P〈0.05）.（4）The ischemic cerebrovascular disease group had significantly higher platelet agglutination rate than the control group（P〈0.01）.The correlation analysis results showed that the platelet agglutination rate in the ischemic cerebrovascular disease group was positively correlated with GPⅠb/Ⅰ Ⅹ/Ⅴ complex expression level（r=0.684）.Conclusion（1）The ischemic cerebrovascular disease patients have a high expression of GPⅠb/Ⅰ Ⅹ/Ⅴcomplex.（2）Ischemic cerebrovascular disease has a susceptibility gene C allele at GPⅠbαkozak site.（3）The increase of platelet agglutination rate is highly correlated with ischemic cerebrovascular disease occurrence.（4）GPⅠbαkozak site gene mutation serves as a genetic factor causing elevating of the GPⅠb/ⅠⅩ/Ⅴcomplex level.（5）The high expression of GPⅠb/ⅠⅩ/Ⅴ complex results in the increase of platelet agglutination rate.