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目的 两个线粒体突变 32 4 3A→G和 3316G→A已经被报道发现在糖尿病患者中 ,本研究旨在调查这个可能的糖尿病相关区域 3116→ 335 3在中国人二型糖尿病中的角色。方法 应用聚合酶链反应 ,直接的DNA测序 ,限制性长度片段多态性和等位特异聚合酶链反应等方法筛查 2 77个二型糖尿病患者和 2 4 1个正常对照的线粒体目的基因区域。对检出的序列改变进行Fisher精确统计分析 ,并应用RNA折叠软件mfold预测 16SrRNA基因区域的变异所引起的最小自由能二级结构改变以确定其功能意义。结果 检出 4个同质体碱基取代 :32 0 0T→C ,32 0 6C→T ,32 90T→Cand 3316G→A。其中 32 0 0T→C只出现在患者中 ,另外三个在患者和正常人中均有检出 ,统计分析显示其频率的不同无显著意义。 32 0 0T→C和 32 0 6C→T位于16SrRNA ,为该研究首次检出 ,前者引起了最小自由能二级结构模型的极大改变 ,而后者几乎没有引起结构变化。结论 本结果建议 32 0 0T→C与二型糖尿病的发展相关 ,而其他几种变异的影响是中性的。不同于日本人的研究 ,3316C→T与二型糖尿病无关。
Purpose Two mitochondrial mutations, 32 4 3A → G and 3316G → A, have been reported in diabetics to investigate the role of this possible diabetes-related region, 3116 → 335 3, in type 2 diabetes in Chinese. Methods PCR-RFLP, direct DNA sequencing, restriction fragment length polymorphism (PCR) and allele-specific polymerase chain reaction (PCR) were used to screen mitochondrial target gene regions in 2 77 diabetic patients and 2,41 normal controls . Fisher exact statistical analysis of the detected sequence changes, and the use of RNA folding software mfold predict 16SrRNA gene mutation caused by the minimum free energy secondary structure changes to determine its functional significance. As a result, four homo-base substitutions were detected: 32 0 0T → C, 32 0 6C → T, 32 90T → Cand 3316G → A. Among them, 32 0 0T → C only appeared in the patients, while the other 3 cases were detected in both patients and normal people. Statistical analysis showed no significant difference in frequency. 32 0 0T → C and 32 0 6C → T in 16S rRNA were the first to be detected in this study. The former caused a great change in the secondary free-energy model of secondary structure, while the latter almost did not cause structural changes. Conclusions This result suggests that 32 0 0T → C is associated with the development of type 2 diabetes while the effects of several other variants are neutral. Unlike Japanese studies, 3316C → T has nothing to do with type 2 diabetes.