Chistyakov D A, Savost'anov K V, Zotova E V, Nosikov V V
INSERM U36, College de France, 75231 Paris Cedex 05, France.
BMC Med Genet. 2001;2:4. doi: 10.1186/1471-2350-2-4. Epub 2001 Mar 28.
Oxidative stress, resulting in a marked increase in the level of oxygen free radicals (OFR), has been implicated in the etiology of diabetic neuropathy (DN). Antioxidant enzymes may protect against the rapid onset and progression of DN, by reducing the excess of OFR and peroxide. Mutations and polymorphisms in the genes encoding such enzymes may therefore result in predisposition to DN. We investigated the role of genes encoding two antioxidant enzymes, mitochondrial (Mn-SOD) and extracellular (EC-SOD) superoxide dismutase, in DN pathogenesis in a Russian population. We studied Ala(-9)Val and Ile58Thr polymorphisms of the Mn-SOD gene and Arg213Gly dimorphism of the EC-SOD gene in type 1 diabetic patients with (n = 82) and without DN (n = 84).
We developed and used a new polymerase chain reaction (PCR) assays for rapid detection of polymorphisms. These assays involved the use of mismatch PCR primers to create restriction sites in the amplified product only in presence of the polymorphic base. The PCR product was than digested with BshTI, Eco32I or Eco52I to detect Ala(-9)Val, Ile58Thr or Arg213Gly polymorphic site respectively. The frequencies of the Ala allele (50.6% vs. 68.5%, p < 0.002) and the Ala/Ala genotype (17.1% vs. 39.3%, p < 0.005) of the Mn-SOD gene were significantly lower in DN patients than in diabetic subjects without DN. In contrast, the Val allele (49.4% vs. 31.5%, p < 0.002) and the Val/Val genotype (15.9% vs. 2.4%, p < 0.01) were significantly more frequent in the DN patients than in the control group.
Ala(-9)Val substitution in the Mn-SOD gene was associated with DN in a Russian population
氧化应激导致氧自由基（OFR）水平显著升高，与糖尿病神经病变（DN）的病因有关。抗氧化酶可通过减少过量的OFR和过氧化物来预防DN的快速发生和进展。因此，编码此类酶的基因中的突变和多态性可能导致易患DN。我们研究了编码两种抗氧化酶，即线粒体（Mn-SOD）和细胞外（EC-SOD）超氧化物歧化酶的基因在俄罗斯人群DN发病机制中的作用。我们研究了1型糖尿病伴DN患者（n = 82）和不伴DN患者（n = 84）中Mn-SOD基因的Ala(-9)Val和Ile58Thr多态性以及EC-SOD基因的Arg213Gly二态性。
我们开发并使用了一种新的聚合酶链反应（PCR）检测方法来快速检测多态性。这些检测方法涉及使用错配PCR引物，仅在存在多态性碱基时在扩增产物中创建限制性位点。然后用BshTI、Eco32I或Eco52I分别消化PCR产物以检测Ala(-9)Val、Ile58Thr或Arg213Gly多态性位点。Mn-SOD基因的Ala等位基因频率（50.6%对68.5%，p < 0.002）和Ala/Ala基因型频率（17.1%对39.3%，p < 立的，没有任何关联。
在俄罗斯人群中，Mn-SOD基因中的Ala(-9)Val替代与DN相关。
