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目的了解肺泡表面活性物质蛋白B(SPB)-18基因多态性与新生儿呼吸窘迫综合征(NRDS)易感性的关系。方法 (1)高分辨率熔解曲线法和基因测序法检测2009年5月至2010年12月武汉市妇女儿童医疗保健中心和华中科技大学同济医学院附属同济医院100例NRDS患儿及186名同胎龄儿SPB-18多态性位点基因型和等位基因的差异。(2)应用蛋白印记法分析技术(WB)检测2009年5月至2010年12月武汉市妇女儿童医疗保健中心31例基因型为SPB-18AA和36例SPB-18CC的足月新生儿支气管肺泡灌洗液中SPB水平,比较两组之间差异。结果 (1)SPB-18A/C基因在NRDS组和对照组中AA、AC、CC的基因型频率分别为11.0%、40.0%、49.0%和6.5%、31.7%、61.8%,两组基因型分布差异无统计学意义(χ2=4.83,P>0.05。);但等位基因A频率在NDRS组和对照组间分别为31.0%和22.3%,差异有统计学意义(χ2=5.19,P<0.05)。(2)基因型为SPB-18AA者比CC者支气管肺泡灌洗液中SPB蛋白水平低。WB灰度值分别为8002.3±452.9和14339.2±1076.3,两组差异有统计学意义(P<0.05)。结论 SPB-18基因多态性是NRDS的危险因素,A等位基因可能通过影响SPB水平,增加NRDS易感性。
Objective To investigate the relationship between alveolar surfactant protein B (SPB) -18 gene polymorphism and susceptibility to neonatal respiratory distress syndrome (NRDS). Methods (1) The high resolution melting curve method and gene sequencing method were used to detect 100 cases of NRDS children and 186 children with Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from May 2009 to December 2010. Differences of genotypes and alleles in SPB-18 polymorphic loci in gestational age children. (2) 31 cases of full-term neonatal bronchial alveoli with genotype SPB-18AA and 36 cases of SPB-18CC were detected by Western blot (WB) from May 2009 to December 2010 in Wuhan Women’s and Children’s Medical Center. SPB levels in lavage fluid were compared between the two groups. Results (1) The frequencies of AA, AC and CC genotypes of SPB-18A / C gene in NRDS group and control group were 11.0%, 40.0%, 49.0% and 6.5%, 31.7% and 61.8% (Χ2 = 4.83, P> 0.05). However, the frequency of allele A was 31.0% and 22.3% in NDRS group and control group, respectively, with significant difference (χ2 = 5.19, P < 0.05). (2) The level of SPB protein in bronchoalveolar lavage fluid of patients with genotype SPB-18AA was lower than that of CC patients. WB gray value were 8002.3 ± 452.9 and 14339.2 ± 1076.3 respectively, the difference between the two groups was statistically significant (P <0.05). Conclusion SPB-18 gene polymorphism is a risk factor for NRDS. A allele may increase the susceptibility to NRDS by affecting SPB level.