脂联素启动子基因多态性与2型糖尿病及并发症的相关性研究

Correlation of polymorphism of adiponectin promoter with diabetes type 2 and its complications

目的观察脂联素（APN）启动子基因单核苷酸多态性（SNP）频率与2型糖尿病（T2DM）及其合并症间的相关性，探讨脂联素基因（aPM1）突变是否是T2DM遗传的危险因素。并确立一种高效、准确、经济的检测APN基因实用筛查方法，供临床检测应用。方法对单纯T2DM49例（男28例，女21例）、T2DM合并高血压（T2DM-HP）90例（男51例，女39例）、T2DM合并冠心病（T2DM-CHD）33例（男15例，女18例）、T2DM合并肾病（T2DM-NE）41例（男18例，女23例）患者和健康对照组58名（男30名，女28名），测定空腹血清生化指标。采用变性高效液相色谱（DHPLC）技术，筛选APN基因多态性位点。结果在APN基因（aPM1）启动子区扩增的片段与基因库中登陆的APN Gene ID：9370序列相比，存在点突变（-11377G／C）。GG、GC、CC基因频率在T2DM组和对照组分别为：5．16％、42．25％、52．58％和3．40％、32．75％、63．85％。T2DM患者aPM，的SNP-11377位点G等位基因频率明显高于健康对照组（r=6．818，OR=0．55，P=0．033）。从T2DM基因型分组临床资料比较可见，-11377G／C基因型与T2DM患者的收缩压（P=0．035）、体重指数（BMI）（P=0．010）、腹围（P=0．013）和腰臀比（P=0．015）显著相关。通过对试验条件优化，在采用DHPLC技术检测APN基因多态性位点发现，本试验检测柱温以60℃最佳。结论aPM，启动子区SNP-11377C／G多态性与T2DM及并发症的发生和T2DM患者体型肥胖形成机制有相关性，提示SNP-11377C／G多态性可能增加T2DM的遗传风险。

Objective To explore the correlation of single nucleotide polymorphism （SNP） frequency of adiponectin（APN） in patients with type Ⅱ diabetes （T2DM） and its complications, investigate whether the SNP is a risk factor of inheritance of T2DM, and to set up a highly efficient, accurate, economical and practical screening assay to detect the mutation of APN in clinical practice. Methods According to the diagnostic criteria of T2DM, patients with coronary heart disease （CHD）, hypertension （liP）, and diabetic nephropathy （NE）were recruited into this study. In simple T2DM group, T2DM-HP, T2DM-CHD, T2DM-NE and the control group, serum biochemistry items are measured. The technique of denaturing high-performance liquid chromatography （DI-IPLC） was used to detect SNPs of ANP gene. Results After all fragments amplified in the region of promoter of APN gene were compared with APN GeneID ： 9370 sequence recorded in GenBank, point mutation has been identified（-11377G/C）. The frequency of genotypes of GG, GC, and CC are 5. 16%, 42. 25%, 52. 58% and 3.4%, 32. 75%, 63. 85%, respectively in the groups of T2DM and control. The frequency of G allele was related to the incidence of T2DM, and is a risk factor of T2DM. The relative risk of GC to CC in developing T2DM is much high than that in the control group （ OR = 0. 55 ）. By comparing the clinical data of different groups of genotype in T2DM, it was observed that the genotype affected systolic blood pressure, BMI, abdominal circumference, and waist-buttock ratio （ P = 0. 015 ）. After optimizing the experimental conditions, it was found column temperature 6 0℃ was the best when using DHPLC technology to estimate SNP of APN gene. Conclusion SNP （ - 11377G/C） of APN gene G allele has a definite correlation with complications of hypertension in T2DM patients, and may contribute to the genetic risk for type 2 diabetes.