【作者】 袁静；

【导师】 丛林；

【作者基本信息】 安徽医科大学 ， 妇产科学， 2008， 硕士

【摘要】 目的:(1)探讨甘露糖结合凝集素(MBL)血清水平与妊娠期糖尿病(GDM)发病的关系,观察MBL基因点突变对GDM发病的影响,初步了解免疫因素在GDM发病中的作用;(2)选择一种检测小片段基因点突变的较好的方法。方法:对2005年9月～2007年3月间在安徽医科大学第一附属医院妇产科接受产前检查的妊娠妇女行GDM的筛查和诊断,检出患GDM的孕妇48人,设为GDM组;在同期健康孕妇中,匹配年龄因素后,随机抽取48人设为健康对照组。所有入组孕妇采集一般的统计学资料,包括年龄、身高、体重、糖尿病家族史等,并于妊娠24-28周抽取静脉血3-5ml,离心后,将血清与细胞成分分开保存待用。采用酶联免疫吸附实验(ELISA)法检测GDM组和健康妊娠组妇女血清中的MBL水平;采用单链多态构象性聚合酶链法(PCR-SSCP)检测两组孕妇MBL基因第54位密码子点突变。运用t检验比较GDM组和健康妊娠组妇女MBL血清水平之间的差异;运用χ~2检验分析MBL血清水平与GDM发病的关系、MBL血清水平与MBL基因第54位密码子点突变的关系以及MBL基因点突变与GDM发病之间的关系;运用多因素非条件Logistic回归分析了解MBL血清水平及MBL基因第54位密码子点突变与GDM发病之间的关联。结果:(1) GDM组与健康妊娠GDM组血清MBL水平呈非正态分布,所测样本的MBL血清水平在4.5 -36.4μg/L之间,平均值14.1±4.4μg/L。GDM孕妇血清MBL的平均值为13.2±6.1μg/L,健康妊娠妇女血清平均值为16.7±5.4μg/L,GDM组孕妇血清MBL的平均值低于健康妊娠妇女,差异有统计学意义,(P＜0.05);(2)以所有孕妇的血清MBL的平均值为界值分组比较结果显示,血清MBL低水平组中,GDM31人,占64.6%(31/48),健康妊娠妇女6人,占12.5%(6/48),低水平组中GDM的发病率明显高于高水平组;GDM组中MBL血清水平在低水平组中的31人,高水平组17人,低水平组高于高水平组,比较具有差异型(χ~2=2.15,P＜0.05);(3)多因素回归分析显示血清MBL水平低(OR=6.5989,95%CI=8.2430～54.6086)、BMI大(OR=4.3852,95%CI=7.3540～34.3561)、有糖尿病家族史(OR=4.1254,95%CI=4.0247～27.2024)均增加孕妇发生妊娠期糖尿病的风险;(P＜0.01):(4)在对PCR产物进行SSCP时发现3种基因型,96例标本中,正常基因型82例,突变型14例;其中GDM组有11例异常,突变率占22.91%;健康妊娠组有3例,突变率占6.25%;GDM组的MBL基因突变率高于健康妊娠组,两组比较结果有差异性。(F=5.62,P＜0.05):(5)多因素Logistic回归分析显示MBL基因点突变(OR=4.971,95%CI=1.094～4.041)增加孕妇发生妊娠期糖尿病的风险(P＜0.01);(6) MBL血清低水平组中有突变10人,占全组的29.4%,血清高水平组中,有突变4人,占全组的6.8%,血清低水平组的突变率明显高于血清高水平组;在有突变的人群中,低水平组10人,高水平组4人,MBL点突变的发生低水平组明显高于高水平组,比较差异有显著性(χ~2=7.17,P＜0.05)。结论:(1) GDM孕妇和健康孕妇的MBL血清浓度存在着差异,本研究显示GDM孕妇的MBL血清水平比健康孕妇低,而低的MBL血清水平更容易引起GDM;(2) MBL基因第54位密码子点突变可能导致MBL血清水平低下,且低水平的MBL容易引起GDM,因此提示MBL基因第54位密码子点突变可能也参与了GDM发病;(3)根据统计学分析显示MBL血清水平和MBL基因突变可能是GDM发病的独立危险因素;(4)PCR-SSCP方法对小片段基因的点突变检测是一种较好的方法。更多还原

【Abstract】 Objective:(1 ) To explore the relationship between MBL serum level and GDM,and to observe the role of point mutation of MBL gene in GDM in order to learn the function of immune factors in GDM;(2) To select a good method to measure short gene point mutation.Methods:Gestational women in prenatal check in The first Affiliated Hospital of Anhui Medical University during Sep.2005 and Mar.2007 were screened and diagnosed in GDM.48 of them were diagnosed with GDM,which were set as GDM group.Among the normal women in the same time,48 were taken out to set as the normal comparison group.General statistics data was collected,including age,stature,weight,family history of diabetes etc.3-5ml venous blood was extracted from gestational women.The serum and cell components of the blood were separately preserved after centrifuged.Enzyme-linked immunosorbent assay was used to measure the MBL serum level of both groups while polymerase chain reaction -Single strand conformation polymorphism(PCR-SSCP) was used to measure the point mutation of the 54th codon of MBL gene.χ~2 est was used to analysis the relationship between the MBL serum level and GDM,the relationship between the MBL serum level and point mutation of the 54th codon of MBL gene,as well as the relationship between the point mutation of the 54th codon of MBL gene and GDM.Non-conditional logistic regression was used to learn the relationship between the MBL serum level and the point mutation of the 54th codon of MBL gene and GDM. Results:(1 ) The serum concentration of MBL in GDM group and normal group had an abnormal distribution.The MBL serum level was between 4.5μg/L and 36.4μg/L,and the average was 14.1±4.4μg/L.The average MBL serum level in GDM was 13.2±6.1μg/L, remarkably lower than that of normal group16.7±5.4μg/L(P＜0.05);(2) It was divided into two groups by the average of MBL serum of all 96 gestational women and the comparison showed that there was 31 women with GDM in low MBL serum level group,while only 6 normal women in this group.GDM morbidity in low MBL level group was evidently higher than the high level group.In GDM group,there was 31 women in low MBL level group,with 17 ones in high level group.There was a significant difference in the comparison of the two groups(χ=2.15,P＜0.05 );(3) The Logistic regression analysis showed that low MBL serum level(OR=6.5989, 95%CI=8.2430-54.6086),high progestation BMI(OR=4.3852,95%CI=7.3540-34.3561 ), family history of diabetes(OR=4.1254,95%CI=4.0247-27.2024) were all increased the risk of GDM(P＜0.01);(4) Three genotypes were found in PCR-SSCP:Among 96 samples,there were 82 normal ones and 14 mutational ones,with 11 mutational ones in GDM group(22.91%) and 3 mutational ones in normal group(6.25%).The mutational rate of MBL gene in GDM group was significantly higher than that of the normal,(5 ) The Logistic regression analysis showed that point mutation of MBL gene (OR=4.971,95%CI=1.094-4.041 ) increased the risk of GDM(P＜0.01);(6 ) There were 10 mutational samples in low MBL serum level group(29.4%).In high serum level group,there were 4 mutational samples(6.8%).The mutational rate of the former group was evidently higher than that of the latter.Among the point mutation women, there was 10 in low MBL serum level group,and 4 in high level group(χ~2=7.17,P＜0.01 ). Conclusions:(1 ) There is difference in MBL serum level between GDM group and the normal group.The study shows that the MBL serum level of GDM is lower than that of normal ones and low serum MBL level can increase the risk of GDM.(2) The point mutation of the 54th codon of MBL gene may lead to the low MBL serum level,and can increase the risk of GDM.This hints that the point mutation of the 54th codon of MBL gene may be one of risk factors of GDM.(3 ) According to the statistical analysis,the MBL serum level and gene point mutation of MBL may be the independent risk factors of GDM.(4) It is better to use PCR-SSCP in point mutation detection of short gene than other methods.更多还原