延边朝鲜族新生儿血红蛋白Gγ/(Gγ+Aγ)比值测定

Determination of Gγ/(Gγ+Aγ) in fetal hemoglobin of Yanbian Korean

目的:血红蛋白γ链包括Gγ和Aγ,基因图谱分析发现新生儿常有γ链基因的异常。实验测定延边朝鲜族新生儿血红蛋白Gγ/(Gγ+Aγ)比值,对于研究人类遗传变异和基因调节具有重要意义。方法:实验于2005-12/2007-04在延边大学医学部基础医学院生物化学与分子生物学教研室完成。①材料来源:97例朝鲜族新生儿脐带血由延边大学医院妇产科产室提供,产妇均签署知情同意书。②实验方法:新生儿脐带血取样后立即冷冻于液氮之中,EDTA-Na2抗凝,进行血红蛋白解链,每500μL脐带血与0.9%的NaCl溶液1000μL相混合稀释,离心去上清,分离血红蛋白。用酸性聚丙烯酰胺以凝胶电泳法分离血红蛋白中的Gγ和Aγ肽链,电压200V,电泳50min,电泳方向为正极到负极。电泳后凝胶用丽春红2R染色,乙酸脱色致本底无色。用UVP扫描凝胶成像分析系统对凝胶板上的Gγ和Aγ肽链进行扫描定量,测定血红蛋白中Gγ/(Gγ+Aγ)比值。结果:朝鲜族新生儿脐带血中的血红蛋白Gγ/(Gγ+Aγ)比值分布情况:1例处于30%～48%低Gγ区,占总例数的1.03%,平均值27.60%;96例处于50%～79%中间区,占总例数的98.97%,平均值(68.40±2.90)%,明显低于相关文献报道的新疆维吾尔族、广西壮族、北京市汉族、西藏藏族新生儿Gγ/(Gγ+Aγ)的整体均值(75.4±2.50)%,差异有显著性意义(P<0.05);未发现>80%高Gγ区者。结论:延边朝鲜族新生儿血红蛋白Gγ/(Gγ+Aγ)比值多分布于50%～79%中间区,其平均值与其他民族比较相对较低。

AIM： The γ chain of hemoglobin （HB） includes Gγ and Aγ. Sometimes abnormal gene can be found in neonates through gene map analysis. In this study, Gγ/（Gγ＋Aγ） ratio in fetal hemoglobin of Yanbian Korean was determined for human research of gene mutation and gene regulation. METHODS： The experiment was conduced in the Department of Biochemistry and Molecular Biology, Yanbian University Health Science Center from December 2005 to April 2007. ①Ninety-seven samples of umbilical cord bloods of Yanbian Korean neonates were obtained from the Department of Obstetrics and Gynecology, Yanbian University Hospitat with the agreement of puerperant. ②The samples were frozen in liquid nitrogen, and EDTA-Na2 was used for anticoagulation. Every 500 μL cord blood was mixed with 1 000 μL NaCI solution （0.9%） and the supernatant was removed by centrifugation to separate hemoglobin. The Gγ and Aγ chains were separated with acid polyacrylamide gel electrophoresis （PAGE） at 200 V for 50 minutes from positive to negative pole. Electrophoresis gel was stained with Lichunhong-2R, destained with acetic acid. The Gγ/（Gγ＋Aγ） ratio was determined by scaning with Imaging System of UVP. RESULTS：①The Gγ（Gγ＋Aγ） ratio of hemoglobin of Yanbian Korean： The ratio of one case was 30%-48% of the low Gγ area, accounting for 1.03% （average 27.60%）, and that of 96 cases was at 50%-79% of the middle area, accounting for 98.97%, average of （68.40＋2.90）%, significantly lower than the general average of Xinjiang Uygur, Guangxi Zhuang, Beijing Han, and Tibet neonates [（75.4.＋.2.50）%, P 〈 0.05]. Moreover, no ratio was above 80%. CONCLUSION： The （Gγ/Gγ＋Aγ） ratio of hemoglobin in Yanbian Korean neonates is distributed in 50%-79% region, and lower than other nations in China.