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目的探讨线粒体DNAtRNALeu(UUR)基因和ND1基因点突变(np3243、3316、3394和3593)与湖北人群2型糖尿病的相关性。方法采用聚合酶链反应限制性片段长度多态性(PCRRFLP),克隆和测序的分析方法,对184例2型糖尿病患者和210名健康对照进行筛选,并用DNAMAN、Primer、mfold和Antherprot软件对检出的突变位点进行分析和二级结构预测。结果实验组检出3316(G→A)突变6例(3.3%),3394(T→C)突变5例(2.7%),3593(T→C)突变1例(0.5%),并发现3个尚未见报道的新突变位点:3606(A→G)、3618(T→C)和3688(G→C),未检出3243(A→G)突变;对照组只检出3316(G→A)突变1例(0.5%)。两组间仅3394(T→C)突变率差异有统计学意义(P<0.05)。3606(A→G)和3618(T→C)突变分别为亮氨酸、苯丙氨酸的无意义突变,其二级结构均与野生型ND1相同;3316(G→A)突变(丙氨酸→苏氨酸),3394(T→C)突变(酪氨酸→组氨酸),3593(T→C)突变(缬氨酸→丙氨酸),3688(G→C)突变(丙氨酸→脯氨酸),均引起ND1基因DNA和蛋白质二级结构的改变,其中3394(T→C)突变型变化最显著。结论线粒体DNAND1基因3394(T→C)突变以及3688(G→C)突变伴随3316(G→A)突变,可能与2型糖尿病的发生发展有关。
Objective To investigate the association between mitochondrial DNA tRNALeu (UUR) gene and ND1 gene point mutations (np3243, 3316, 3394 and 3593) and type 2 diabetes in Hubei population. Methods 184 patients with type 2 diabetes and 210 healthy controls were screened by polymerase chain reaction-restriction fragment length polymorphism (PCRRFLP), cloning and sequencing. DNAMAN, Primer, mfold and Antherprot software Out of the mutation sites for analysis and secondary structure prediction. Results In the experimental group, 6 cases (3.3%) of 3316 (G → A), 5 (2.7%) 3394 (T → C) and 1 (35% No new mutation sites were reported in this study: 3606 (A → G), 3618 (T → C) and 3688 (G → C), no 3243 (A → G) mutation was detected; → A) Mutation in 1 case (0.5%). Only 3394 (T → C) mutation rate between the two groups was statistically significant (P <0.05). The secondary mutations of 3606 (A → G) and 3618 (T → C) were leucine and phenylalanine, respectively. The secondary structure was the same as that of wild type ND1. The mutation of 3316 (G → A) Acid → threonine), 3394 (T → C) (tyrosine → histidine), 3593 (T → C) (valine → alanine), 3688 Amino acid to proline), all caused the DNA and protein secondary structure changes of ND1 gene. Among them, the 3394 (T → C) mutant had the most significant change. Conclusion The mutations of 3392 (T → C) and 3688 (G → C) mutations of mitochondrial DNAND1 gene accompanied with 3316 (G → A) mutation may be related to the development of type 2 diabetes mellitus.