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目的探讨1型糖尿病(T1DM)酮症酸中毒(DKA)患儿缺氧诱导因子-1α(HIF-1α)与血管内皮细胞生长因子(VEGF)mRNA水平的变化。方法天津市儿童医院住院T1DM并DKA患儿30例,于确诊24 h内(DKA 1组)及DKA纠正后10 d(DKA 2组)采血,另选取同期住院的不伴感染、缺氧、肿瘤或结缔组织病的同年龄同性别患儿30例为对照组。实时荧光定量PCR(Real-time PCR)法测定其外周血CD4+T淋巴细胞HIF-1α与VEGF mRNA的相对表达水平。PCR产物行琼脂糖凝胶电泳鉴定特异性。采用SPSS 13.0软件进行统计学分析。结果 3组HIF-1α及VEGF mRNA相对表达水平比较差异均有统计学意义(Pa<0.01)。DKA1组HIF-1α及VEGF水平明显高于对照组,差异有统计学意义(Pa<0.01);DKA纠正后HIF-1α及VEGF水平恢复,差异有统计学意义(P<0.05,0.01),但直至DKA纠正后10 d(DKA2组)仍未恢复至对照组水平,差异有统计学意义(P<0.01,0.05)。Real-timePCR产物行琼脂糖凝胶电泳,产物位于预期位置,确定产物特异性。结论 T1DM并DKA患儿CD4+T淋巴细胞HIF-1α与VEGFmRNA水平升高,且DKA纠正后HIF-1α与VEGF mRNA水平不能恢复至正常,这可能与T1DM并发症的发生发展有关。
Objective To investigate the changes of hypoxia inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) mRNA in type 1 diabetic ketoacidosis (DKA) patients. Methods Thirty children with T1DM and DKA were admitted to Tianjin Children ’s Hospital. Blood samples were collected within 24 hours of diagnosis (DKA 1) and 10 days after DKA correction (DKA 2). Other hospitalized patients without infection, hypoxia and tumor Or connective tissue disease of the same age and 30 children with the same sex as the control group. The relative expression levels of HIF-1α and VEGF mRNA in peripheral blood CD4 + T lymphocytes were measured by real-time PCR. PCR products were identified by agarose gel electrophoresis. SPSS 13.0 software was used for statistical analysis. Results The relative expression levels of HIF-1α and VEGF mRNA in the three groups were significantly different (Pa <0.01). The levels of HIF-1α and VEGF in DKA1 group were significantly higher than those in control group (P <0.01), while the levels of HIF-1α and VEGF in DKA1 group were recovered (P <0.05, 0.01) Until DKA corrected 10 d (DKA2 group) has not recovered to the control group, the difference was statistically significant (P <0.01, 0.05). Real-timePCR products were subjected to agarose gel electrophoresis and the product was located at the expected position to confirm product specificity. Conclusions The levels of HIF-1α and VEGFmRNA in CD4 + T lymphocytes of T1DM and DKA patients are elevated, and the levels of HIF-1α and VEGF mRNA can not recover to normal after DKA correction. This may be related to the occurrence and development of T1DM complications.