【作者】 邢明青；

【导师】 罗兵； 曲彦； 纪霞；

【作者基本信息】 青岛大学 ， 病原生物学， 2003， 硕士

【摘要】 目的 纤维蛋白原激活剂抑制剂-1(plasminogen activator inhibitor，PAI-1)是一种单链糖蛋白，属于丝氨酸蛋白酶抑制剂家族成员。纤维蛋白溶解过程的关键步骤是纤维蛋白溶解酶原在纤维蛋白溶解酶原激活剂(plasminogen activator，PA)作用下转变为纤维蛋白溶解酶，使纤维蛋白溶解，PAI-1的主要功能是快速、有效的抑制PA，导致纤维蛋白溶解作用减弱，血栓形成。血管紧张素转化酶(angiotensin converting enzyme，ACE)是一种含锌的膜结合蛋白，属二肽羧肽酶，能使血管紧张素Ⅰ转化为血管紧张素Ⅱ，后者通过与血管、气管平滑肌的作用，引起血管、支气管收缩。临床资料表明肺心病患者多伴有高凝状态和血管紧张，本研究旨在通过对肺心病患者ACE和PAI-1基因多态性的检测，从分子水平探讨肺心病的发病机理，寻找肺心病发病的相关致病基因。 方法 应用多聚酶链反应(PCR)，检测40例正常人和60例肺心病患者ACE、PAI-1的基因型和等位基因频率。 结果 ①正常对照组ACE基因型分布为DD型10.0％，II型47.5％，ID型42.5％；肺心病组ACE基因型分布为DD型20.0％，II型36.7％，ID型43.3％。统计学分析表明两组间基因型构成、I和D等位基因频率以及DD型与非DD型构成均无显著性差异。②正常对照组PAI-1基因型分布为4G／4G型12.5％，5G／5G型15.0％，4G／5G型72.5％；肺心病组PAI-1基因型分布为4G／4G型15.0％，5G／5G型28.3％，4G／5G型56.7％。统计学分析表明两组间基因型构成、4G和5G等位基因频率以及4G／4G型与非4G／4G型构成均无显著性差异。③正常女性组中ACE基因型分布为DD型12.5％，ID型37.5％，II型50.0％，等位基因频率D型为31.2％，I型为68.8％。男性组中ACE基因型分布为DD型8.4％，ID型45.8％，II型45.8％，等位基因频率D型为31.2％，I型为68.8％。统计学分析显示两组间基因型构成、等位基因频率以及DD型与非DD型构成无显著性差异。④女性肺心病患者ACE基因型分布DD型25.9％，II型48.2％，ID型25.9％；男性肺心病患者ACE基因型分布为DD型15.1％，II型27.3％，ID型57.6％．统计学分析表明两组间基因型构成有明显差异，I／D等位基因频率以及DD型与非DD型基因型构成则无明显差异。⑤正常对照组女 中文摘要性组PAI一1基因型分布为4G/4G型6 .2%，4G/SG型68.8%，SG/SG型25.0环;等位基因频率4G型为40.6%，SG型为59.4%;男性组中4G/4G型16.7写，4G/SG型75.0%，SG/SG型8.3%，等位墓因频率4G型为54.2%，SG型为45.8%.统计学分析表明两组间基因型构成、等位墓因频率以及4G/4G型与非4G/4G型构成均无显著性差异。⑥女性肺心病患者PAI一1墓因型分布为4G/4G型25.9%，4G/SG型55.6%，SG/SG型18.5%;男性肺心病患者PAI一1基因型分布为4G/4G型6.1写，4G/SG型57.6%，SG/5G型36.3%.统计学分析表明两组间基因型构成无显著性差异，4G/SG等位基因频率有显著性差异，4G/4G型与非4G/4G型基因型构成无显著性差异。⑦在肺心病伴有肺栓塞组的19例患者中ACE基因DD型、ID型和11型分别为3 1 .6%、42.1%和26.3%，等位基因频率D型为52.6%，I型为47.4%.统计学分析表明，与正常对照组基因型构成比较无显著性差异，DD型与非DD型之间有显著性差异，D/I等位基因频率有显著性差异。⑧在肺心病伴有肺栓塞组的19例患者中PAI一1墓因4G/4G型、4G/SG型和SG/SG型分别为10.5%、68.4写和21.1%，等位基因频率4G型为44.7%，SG型为55.3%.统计学分析表明，与正常对照组墓因型构成比较、4G/4G型与非4G/4G型之间以及4G/5G等位基因频率均无显著性差异。⑨肺心病组、DD基因型患者组和4G/4G基因型患者组的Hct值范围分别为。.521士。.155、0.528士0.112和0.519士0.097.它们分别与对照组的Hct值0.483士0.056进行组间均数比较，显示3组间均无统计学意义(P>0.05). 结论①ACE括入/缺失墓因多态性与肺心病无明显相关性，DD基因型不是肺心病发病的危险因子，然而DD墓因型在肺心病组有增高的趋势。②PAI一l4G/SG墓因多态性与肺心病无明显相关性，4G/4G基因型不是肺心病发病的危险因子。③肺心病患者中女性ACE基因中的DD基因型有升高趋势，DD基因型可能是女性患肺心病的危险因子。④肺心病患者中，4G等位基因可能是女性患肺心病的危险因子。⑤DD基因型与肺心病伴肺栓塞有显著相关性，DD墓因型和D等位基因可能是肺心病伴肺梗塞的危险因子;4G/4G基因型和4G等位基因与肺心病伴肺栓塞无明显相关性。⑥DD基因型和4G/4G基因型患者与正常对照组平均红细胞压积无明显相关性。更多还原

【Abstract】 Objactive Plasminogen activator inhibitor-1 (PAI-1), a kind of glycoprotein, is a member of the SERPIN family. The important step in the progress of fibrinolysis is that tissue plasminogen activator(PA) transforms plasminogen into plasmin, the latter makes fibrin degradation. PAI-1 main function is as the fast-acting inhibitor of t-PA. This action prevents the development of fibrinolysis and makes thromsis. Angiotensin converting enzyme (ACE), membrance binding protein, is a member of peptidyl-dipeptide hydrolase. ACE transforms angiotensin I into angiotensin II. Ang II produces construction of vascular and trachea by acting on smooth muscle. The clinical studies show that most patients of chronic pulmonary heart disease(CPHD) have the syndrom of high risk blood clot and an-giotonia. We investgates the reason and relative gene of CPHD on molecular level by detecting the polymorphism of ACE gene and PAI-1 gene.Methods The ACE and PAI-1 genotype in 40 normal subjects and 60 pateints of CPHD is detected by the polymerase chain reaction (PCR).Results (1)In the control group t the distribution of the DD, II and ID genotypes of the ACE gene is 10. 0%,47. 5% and 42. 5%;In the study group, the distribution of the DD,II and ID genotypes of the ACE gene is 20. 0%,36. 7% and 43. 3%; These percentages in control group do not significantly differ from the study group, I/Dallele frequency and DD genotype /non- DD genotype in control group do not significantly differ from the study group. (2)In the control group, the distribution of the 4G/4G,4G/5G and 5G/5G genotypes of the PAI-1 gene is 12. 5% ,72. 5% and 15. 0%;In the study group, the distribution of the 4G/4G,4G/5G and 5G/5G genotypes of the PAI-1 gene is 15. 0%,56. 7% and 28. 3 %; These percentages in control group do not significantly differ from the study group, 4G allele frequency and 4G/4G genotype /non- 4G/4G genotype in control group do not significantly differ from the study group. (3)In the female subgroup of CPHD, the distribution of the DD,II and ID genotypes of the ACE gene is 25. 9% ,48. 2% and 25.9%; In the male subgroup, the distribution of the DD,II and ID genotypes of the ACE gene is 15.1%,27. 3% and 57. 6%;These percentages in female subgroup significantly differ from the male subgroup, I/D allele frequency and DD genotype /non- DD genotype in femalesubgroup do not significantly differ form the male subgroup. (4)In the female subgroup of CPHD, the distribution of the 4G/4G.4G/5G and 5G/5G genotypes of the PAI-1 gene is25. 9% ,55. 6% and 18. 5%; In the male subgroup, the distribution of the 4G/4G.4G/5G and 5G/5G genotypes of the PAI-1 gene is 6. 1%, 57. 6% and 36. 3%;These percentages and 4G/4G genotype /non- 4G/4G genotype in female subgroup do not significantly differ from the male subgroup, 4G allele frequency in female subgroup significantly differ from the male subgroup. (5)In 19 subjects of patients of CPHD accompanying with pulmonary embolism, the distribution of the DD.II and ID genotypes of the ACE gene is 31. 6%,26. 3% and 42. 1%. D/I allele frequency is 52. 6% and 47. 4%. The percentages of ACE genotypes is not significantly different in control group and subgroup, The percentages of DD genotype and non-DD genotype, the I/D alleles frequency are significant association between the two groups. (6)In 19 subjects of patients of CPHD accompanying with pulmonary embolism, the distribution of the 4G/4G,5G/5G and 4G/5G genotypes of the PAI-1 gene is 10. 5 % ,21. 1 % and 68. 4 %. 4G/5G allele frequency is 44. 7 % and 55. 3 %. The percentages of PAI-1 genotypes, the percentages of 4G/4G genotype and non-4G/4G genotype, the 4G/5G alleles frequency are not significantly different. (7)The data of mean Hct is 0. 521±0. 155, 0. 528±0. 112 and 0. 519 ± 0. 097 in the CPHD group, DD genotype subgroup and 4G/4G genotype subgroup. In the control group, the mean Hct is 0. 483 ± 0. 056. There is not significantly different in control group comparing with CPHD group, DD genotype subgroup and 4G/4G genotype subgroup.Conclusion (1)更多还原