| Background:Subarachnoid hemorrhage (SAH) is an acute onset, rapid development, high mortality and morbidity’s serious neurosurgical diseases, aneurysms, arteriovenous malformations and atherosclerosis are the common causes. Apolipoprotein (a) [Apo(a)] is one of the major components of Lipoprotein(a) [Lp(a)], its coding gene [Apo(a) gene] containing Kringle IV-2 [KIV-2] repeat copy number polymorphisms, (TTTTA)n polymorphism, single nucleotide polymorphisms (SNPs) and others rich gene polymorphisms. Apo(a) gene polymorphism is closely related to the occurrence of vascular diseases, such as atherosclerosis, ischemic stroke, intracranial aneurysms,etc. And determined more than 90 percentage of the variance of the level of plasma Lp(a). But so far, there were no reports about the association between Apo(a) gene polymorphism and the incidence of SAH at home and abroad.Object:To study the influence of Apo(a) gene polymorphism on the risk of SAH and plasma Lp (a) levels variety.Method:Collect 32 cases of SAH patients from neurosurgery Affiliated Hospital of Kunming University of hospital treatment as the experimental group,36 cases healthy control subjects were collected from Kunming University Affiliated Hospital Medical Center. Extracting venous blood in the early morning, centrifugal separating plasma and blood cell, detected and analyzed plasma total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), apolipoprotein AI (ApoAI), apolipoprotein B (ApoB) and lipoprotein (a) [Lp (a)] levels, and compared the difference between the two groups. Polymerase chain reaction (PCR) combined with polyacrylamide gel electrophoresis (PAGE), western blot (WB) and other technologies been used to detected the polymorphism in the 5’regulatory region [+914 site A→G,+49 site C→T and (TTTTA)n], the first exon sequence polymorphism of Apo(a) [Apo(a)-1], the intron sequence polymorphisms between two exons which encoding the first KIV-2 encoding duplicate (KIV-2-1) of Apo(a) gene, as well as the protein phenotype polymorphism of Apo(a) (ie. KIV-2 copy number repeat polymorphism). What’s more, the effect of these gene polymorphism on the level of plasma Lp(a) have been analyzed.Result:SAH patients plasma level of Lp(a) (186.19±180.54mg/L) is significantly higher than healthy subjects (92.64±61.54mg/L) (P< 0.05); Compared the levels of TC (4.30±0.99mg/L:4.21±1.07mg/L), TG (1.35±0.77mg/L:1.89±1.58mg/L), HDL-C(1.13±0.32mg/L:1.00±0.39mg/L),LDL-C (2.59±0.73mg/L:2.50±0.76mg/L), ApoAI(1.17±0.24mg/L:1.11±0.34mg/L) and ApoB (0.94±0.21mg/L:0.97±0.32mg/L) between the two groups, no significant difference have been found; there were no correlation between plasma Lp(a) levels and TC, TG, HDL-C, LDL-C, ApoAI and ApoB. The+49 site C→T polymorphisms of Apo(a) gene analysis were detected a and β two genotypes altogether, and significant genotype frequency distribution difference have been found between the two groups (x 2=19.833, P< 0.05). Experimental groups genotype a carriers plasma Lp(a) levels were significantly higher than control groups genotype a carriers (279.78±93.53mg/L:92.64±32.54mg/L, P<0.05). Within experimental groups, genotype a carriers the level of plasma Lp(a) was significantly higher than genotype β carriers (279.78±93.53mg/L:65.86±21.36mg/L, P<0.05). Apo(a) gene (TTTTA)n polymorphism analysis were detected three alleles [(TTTTA)3, (TTTTA)5 and (TTTTA)6]and four genotypes [(TTTTA)3/3, (TTTTA)3/5, (TTTTA)3/6 and (TTTTA)6/6] in total in the experimental group and the controls group. Between the two groups, alleles frequency distribution have significant difference (x 2=8.405, P＜0.05), but no significant difference had been found in genotypes frequency distribution (χ2=7.303, P> 0.05). Compared the alleles related plasma Lp(a) levels between the two groups, we found that allele (TTTTA)3, (TTTTA)5 and (TTTTA)6 related Lp(a) levels in the experimental group higher than the controls group; among the alleles, the allele (TTTTA)3 related plasma Lp(a) levels in the experimental group significantly higher than the control group (335.45±217.45mg/L:168.82±54.79mg/L, P< 0.05); genotype (TTTTA)3/5 and (TTTTA)6/6 related Lp(a) levels in the experimental group were slightly higher than the control group, genotype (TTTTA)5/6 related Lp(a) levels in the experimental group was slightly lower than the control group, but no significant defference (P< 0.05). Each alleles related Lp(a) levels both in the experimental group and the control group were significantly different (F= 13.088, P＜0.05; F=46.320, P<0.05), allele (TTTTA)6 related with the lowest Lp(a) levels; there were significant defference each genotype related Lp(a) levels both in the experimental group and the control group (F=76.197, P<0.05; F=116.567, P<0.05), genotype (TTTTA)6/6 correlated with low Lp(a). KIV-2-1 polymorphism of the Apo(a) were detected TD-3 and TD-5 two genotypes altogether in our study, and significant frequency distribution defference has been found between the experimental group and the control group (P< 0.05); genotype TD-3-associated (235.90±205.29mg/L:168.82 ±54.79mg/L) and TD-5-associated (103.33±84.12mg/L:59.12±21.04mg/L) plasma Lp(a) levels in the experimental group higher than the control group, but no significant difference(t=1.375, P> 0.05; t=1.794, P> 0.05). TD-5 carrers plasma Lp(a) levels were significantly lower than genotype TD-3 carrers both in the experimental group and the control group (103.33±40.12mg/L:235.90±205.29mg/L, t=2.553, P< 0.05; 59.12±21.04mg/L:168.82±54.79mg/L, t=6.435, P< 0.05). Apo(a) protein phenotype polymorphism (ie. KIV-2 copy number repeat polymorphism) analysis were detected F and B/F two kinds of Apo(a) isoforms, and no significant frequency distribution difference between the experimental group and the control group (χ 2=1.594, P> 0.05); compared with the control group, Apo(a) isoforms F and B/F-associated plasma Lp(a) levels in the experimental group were higher (239.20± 197.08mg/L:140.29 ± 58.88mg/L; 97.83±105.54mg/L:50.00±14.78mg/L), and Apo(a) isoforms F-associated plasma Lp(a) levels was significantly higher than the control group (t=2.135, P< 0.05). F-type Apo(a) isoforms related plasma Lp(a) levels were significantly higher than B/F type Apo(a) isoforms both in the experimental group and the control group (239.20±197.08mg/L:97.83±105.54mg/L, t=2.635,P< 0.05;140.29±58.88mg/L:50.00±14.78mg/L, t=6.151, P<0.05).Conclusion:Increased plasma Lp(a) level is associated with the increase of the risk of SAH, but influenced by the Apo(a) gene polymorphism;+49 site C→T and KIV-2-1 polymorphism of the Apo(a) gene may associated with the development risk of SAH, and impact of plasma Lp(a) levels variety; Apo(a) gene (TTTTA)n polymorphism may not related with the risk of SAH, but have an effect on plasma Lp(a) levels change. |
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