【摘要】 目的 探讨广西环江地区新生儿葡萄糖-6-磷酸脱氢酶(G6PD)基因突变类型，以及G6PD缺乏与胆红素-尿苷二磷酸葡萄糖醛酸转移酶1A1(UGT1A1)基因编码区第1外显子区G71R错义突变对新生儿高胆红素血症严重程度的影响。方法 回顾性分析2013年1月至2016年12月在环江毛南族自治县人民医院因新生儿高胆红素血症住院治疗的150例新生儿的临床资料。采用酶活性测定法检测高胆红素血症新生儿的G6PD酶活性，将其分为G6PD缺乏组(n=40)与对照组(n=110),对新生儿血标本进行全基因组DNA提纯后，将UGT1A1基因编码区第1外显子区进行PCR扩增、凝胶电泳、基因测序，对G6PD缺乏组采用PCR-限制性酶切分析(PCR/REA)明确G6PD基因突变类型，并将结果进行统计学分析。结果 G6PD缺乏组血清总胆红素水平明显高于对照组，差异有统计学意义(t=2.360,P<0.05)。G6PD基因突变类型分别为G1388A(17例)、G1376T(11例)、A95G(5例)、C1024T(4例),有3例未定型。G6PD基因不同突变类型间血清总胆红素水平比较差异无统计学意义(P>0.05)。两组在UGT1A1基因编码区第1外显子区存在G71R错义突变，其G71R错义突变等位基因频率分别为16.3%、18.6%。G6PD缺乏组的G71R错义突变基因型分布及等位基因频率与对照组比较差异均无统计学意义(P>0.05)。两组的UGT1A1 G71R突变型和UGT1A1 G71R野生型血清总胆红素水平比较差异有统计学意义(F=5.584,P<0.05)。G6PD缺乏组+UGT1A1 G71R突变型、G6PD缺乏组+UGT1A1 G71R野生型、对照组+UGT1A1 G71R突变型的血清总胆红素水平均高于对照组+UGT1A1 G71R野生型，差异均有统计学意义(t值分别为3.256、2.801、3.178,P<0.05)。结论 广西环江地区G6PD缺乏常见突变类型为G1388A、G1376T、A95G和C1024T,G6PD基因不同突变类型之间对血清总胆红素水平的影响无差异。G6PD缺乏与UGT1A1 G71R错义突变对该地区新生儿高胆红素血症的严重程度有协同作用。更多还原

【Abstract】 Objective To investigate mutation types of glucose-6-phosphate dehydrogenase(G6PD) gene, and effects of G6PD deficiency and G71R missense mutation in the first exon area of bilirubin-uridine diphosphate glucuronosyltransferase 1A1(UGT1A1) gene coding region on severity of neonatal hyperbilirubinemia in Huanjiang Maonan Nationality Autonomous County of Guangxi province.Methods The clinical data of 150 neonates with neonatal hyperbilirubinemia who admitted to The People′s Hospital of Huanjiang Maonan Nationality Autonomous County over a period from January 2013 to December 2016 were retrospectively analyzed.G6PD enzyme activity of 150 neonates with hyperbilirubinemia was detected by using enzyme activity assay method.According to the enzyme testing results, the neonates were divided into G6PD deficiency group(n=40) and non-G6PD deficiency group(control group)(n=110).After whole genome DNA purification of the neonatal blood samples in the two groups, PCR amplification, gel electrophoresis and gene sequencing on the first exon area of the coding region of UGT1A1 gene were performed, and PCR-restriction endonuclease analysis(PCR/REA) was used to determine types of G6PD gene mutation of the neonates in the G6PD deficiency group.The results were statistically analyzed.Results The mutation types of G6PD gene were G1388A(n=17),G1376T(n=11),A95G(n=5) and C1024T(n=4).There were 3 cases undefined.There was no significant difference in serum total bilirubin level among different mutation types of G6PD gene subgroups(P>0.05).The serum total bilirubin level of the neonates in the G6PD deficiency group was significantly higher than that in the control group, and the difference was statistically significant(t=2.360,P<0.05).There were G71R missense mutations in the first exon area of UGT1A1 gene coding region of the neonates in the two groups, and the allele frequencies of G71R missense mutations were 16.3% and 18.6% respectively.There were no significant differences in genotype distribution and allele frequency of G71R missense mutations of the neonates between the G6PD deficiency group and the control group(P>0.05).While there were significant differences in serum total bilirubin levels between UGT1A1 G71R mutant type and UGT1A1 G71R wild type of the neonates in the both two groups(F=5.584,P<0.05).The serum total bilirubin levels of the neonates in the G6PD deficiency + UGT1A1 G71R mutant type subgroup, G6PD deficiency + UGT1A1 G71R wild type subgroup and non-G6PD deficiency + UGT1A1 G71R mutant type subgroup were higher than those in the non-G6PD deficiency + UGT1A1 G71R wild type subgroup, and the differences were statistically significant(t=3.256,2.801 and 3.178 respectively, all P<0.05).Conclusion The common mutation types of G6PD deficiency gene in Huanjiang Maonan Nationality Autonomous County of Guangxi are G1388A,G1376T,A95G and C1024T,and there are no differences in effects of G6PD gene on serum total bilirubin level among different mutation types.G6PD deficiency and UGT1A1 G71R missense mutation have synergistic effects on severity of neonatal hyperbilirubinemia in this area.更多还原