【作者】 孙红；

【导师】 刘昭前；

【作者基本信息】 中南大学 ， 临床药理， 2007， 硕士

【摘要】 目的：查明脂联素基因SNP-11377(C／G)和SNP45(T／G)在中国汉族2型糖尿病(T2D)患者和健康对照者中的分布频率，并了解中国汉族2型糖尿病人群中脂联素基因多态性对罗格列酮降糖疗效的影响，为临床实施2型糖尿病个体化药物治疗提供实验依据。方法：应用PCR-RFLP方法对受试者进行脂联素基因常见突变位点SNP-11377(C／G)和SNP45(T／G)基因分型，单倍型频率采用Phase软件计算。从已经完成基因分型的2型糖尿病患者中，选取42个不同基因型的患者每天早饭时口服罗格列酮4mg，连续服药12周。应用单因素方差分析、卡方检验、配对t检验及非参数检验进行统计学分析。结果：对255例T2D患者及其120例健康对照者进行了基因分型，发现其等位基因的发生频率符合Hardy-Weinberg平衡。2型糖尿病病人脂联素基因-11377(C／G)和45(T／G)发生频率分别为28.8％和28.0％，与健康人群频率相比，SNP-11377等位基因频率与基因型频率均存在显著性差异，并且随着G等位基因的增加，患者的脂联素水平(P＜0.001)明显降低，FINS(P＜0.05)和HOMA-IR水平(P＜0.05)明显增高。T2D病人SNP45(T／G)等位基因频率与基因型频率与健康人相比无显著性差异，在糖尿病人群中还发现SNP45 T／T基因型的腰臀比明显高于突变组(P＜0.05)。近一步分析发现-11377(C／G)和45(T／G)存在连锁不平衡，在T2D病人中其携带二倍体CGTT的患者的FPG、PPG、FINS、HOMA-IR及LDL-c明显高于其他二倍体的患者(P＜0.05)。最后我们分析了脂联素基因多态性对罗格列酮疗效的影响，发现对于SNP-11377(C／G)，与突变型相比，携带CC型患者的FPG(P＜0.001)、PPG(P＜0.05)和HOMA-IR(P＜0.05)明显降低，而脂联素水平也是显著升高(P＜0.05)。对于SNP45(T／G)，与TT基因型相比，携带G等位基因患者用药后脂联素水平显著升高(P＜0.05)。对于SNP-1377及SNP45组成的二倍体中，与11／12(CGTT)相比，携带其他二倍体者用药后FPG(P＜0.01)、PPG(P＜0.05)和HOMA-IR(P＜0.05)显著降低，而脂联素水平(P＜0.01)也是显著升高。结论：研究结果揭示脂联素-11377(C／G)突变和携带CGTT二倍体的患者与胰岛素抵抗和2型糖尿病密切相关，可能增加个体患2型糖尿病的危险性；SNP45 T／G变异与肥胖度相关。研究还发现在2型糖尿病中SNP-11377携带CC基因型者对罗格列酮疗效敏感，而CGTT二倍体者对罗格列酮反应差。因此，脂联素基因的变异可能影响罗格列酮的降糖疗效、同时具有改善胰岛素抵抗和升高脂联素水平的作用。更多还原

【Abstract】 AIMS: In this study, we were to investigate the distribution frequency of adiponectin of single nucleotide polymorphisms (SNP) and its haplotypes and determine effect of genetic polymorphisms of adiponectin on the therapeutical efficacy of rosiglitazone in patients with type 2 diabetes(T2D).METHODS: A totle of 255 T2D patients and 120 health volunteers were selected to genotype at -11377(C/G) and +45(T/G) polymorphisms of adiponectin gene by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. Samples were randomly selected to direct sequencing to identify the results of PCR-RFLP. 42 T2D patients were selected to administere orally 4mg rosiglitazone daily for 12 consecutive weeks. Plasma concentration of FPG, PPG, HbAlc as well as serum concentration of FINS and PINS in all subjects were determined at 0 and 12th weekend. The frequency of haplotypes was calculated using phase sofeware haplotypes estimation. Statistical analyses include one-way ANOVA, paird t test ,Chi-square (?~2) analysis and non-parametric test.RESULTS: The distribute characters of both alleles were accorded with Hardy-Weinberg equilibrium. In T2D, the allele frequencies for-11377(C/G) and +45(T/G) were 28.8% and 28.0% respectively. There were significant different in allele frequency of-11377(C/G) between health volunteers and T2D patients.However, in subjects with -11377(C/G) genotypes, there was a significant lower in the serum adiponectin concentration (P＜0.001) and higher in the FINS(P＜0.05 ) and HOMA-IR(P＜0.05 ) for the G allele gene than CC genotype in T2D patients (P＜0.05). In subjects with 45(T/G) genotypes, there was a marked higher in the waist-to-hip ratio in patients with the TT genotype than those with the others genotype in T2D patients (P＜ 0.05). To haplotype analysis, the level of FPG(P＜0.05), PPG(P＜0.01), FINS(P＜0.05) and LDL-c(P＜0.05) were higher in those with CGTT diplotype than those with the other diplotypes (P＜0.05). Finally, we observed the effect of C-11377G and T45G polymorphisms on the therapeutic efficacy of rosiglitazone in patients with T2D. In subjects with -11377 genotypes, the degree of the decrease in the FPG (P＜0.001), PPG (P＜0.05) and HOMA-IR (P＜0.05) level were significant higher in those with the CC genotype than those with the other genotypes.However, the degree of the increase in the serum adiponectin concentration(P＜0.01) was significantly higher in those with the CC genotype than those with the other genotypes.In subjects with 45 genotypes, the degree of the increase in the serum adiponectin concentration was significantly s in those with the G allele than those with the CC genotype (P＜0.05). To haplotype analysis, the degree of the reduction in the FPG FPG (P＜0.001), PPG (P＜0.05) and HOMA-IR (P＜0.05) level was smaller for the CGTT diplotype than for the other diplotypes. In addition, the degree of the increase in the adiponectin concentration(P＜0.001) was significant smaller in the subjects with CGTT diplotype than the subjects with the other diplotypes.CONCLUSION: -11377(C/G) and +45(T/G) are common polymorphisms of adiponectin gene in Chinese population. There was an associated between polymorphism of adiponectin gene -11377(C/G) and CGTT diplotype with the pathogenesis of T2D. The -11377G allele and CGTT diplotype might be a risk factor for the development of T2D. This study suggests that patients with CC genotype at locus -11377 are sensitive to the therapeutical efficacy of rosiglitazone in patients with type 2 diabetes. However, patients with CGTT diplotype are unlikely to respond to the rosiglitazone.These data suggest that genetic variations in the adiponectin gene could affect the therapeutic efficacy and improve the insulin resistance and increase the level of adiponectin in patients with type 2 diabetic patients.更多还原