μ阿片受体A118G基因多态性与早产儿颅内出血的相关性研究

发布时间：2018-08-04 19:44

【摘要】：背景及目的颅内出血（Intracranial hemorrhage，ICH）是新生儿常见的一种严重颅脑损伤，早产儿多见，胎龄越小，发病率越高，严重者可留有神经系统后遗症，甚至死亡。近年来，随着医学技术的不断发展，早产儿存活率大大提高，ICH的发生率有增无减。 ICH的发生与自身的解剖生理特点和多种围产期高危因素有关，此外，许多炎性反应和内源性介质等也可诱发，其中内源性神经介质特别是内源性阿片肽的释放增加能加重这种损伤。近年来，神经肽与脑血管疾病机理研究日益受到关注,研究发现阿片肽及阿片受体系统在成人缺血性脑卒中的病理生理发展过程中具有非常重要的作用。临床研究发现有着相同危险因素的早产儿ICH的发生存在明显的个体差异，基因遗传学背景是造成个体差异性的重要原因之一。前期研究发现血小板活化因子PAF-AH基因Val279Phe单核苷酸多态性与早产儿ICH具有相关性，我们在此基础上探索ICH发生的其他易感基因。 μ阿片受体(μ opioid receptor，OPRMI)A118G基因多态性与多种疾病的易感性及药物反应的差异性有关，而其与新生儿疾病的相关性研究目前尚未见报道。本研究拟选择被大家广泛认可的与精神神经因素密切相关的OPRMIA118G单核苷酸基因多态性，研究其与早产儿颅内出血的关联性，探讨ICH发生的分子遗传学机制，为临床有效防治ICH提供参考依据。材料和方法 1研究对象与分组选取2011.7至2013.3郑州大学第一附属医院新生儿重症监护室(NICU)住院的汉族早产儿作为研究对象。将颅内出血早产儿视为颅内出血组，同期因早产要求住院观察的非颅内出血早产儿视为非颅内出血组。并对颅内出血进行分度。2基因多态性检测采用聚合酶链式反应一限制性片段长度多态性分析(Polymerase chainreaction restriction fragment length polymorphisms，PCR-RFLP)技术，对OPRMI基因Al18G多态性位点进行检测分析， PCR产物直接测序验证基因型检测方法的可靠性。3统计学分析采用SPSS19.0统计软件进行数据统计分析，两组研究对象胎龄、出生体质量之间的比较采用独立样本t检验，性别构成的比较检验、Hard-Weinberg平衡法则检验、各组基因型及等位基因分布的比较、性别及出血程度与颅内出血的关联性比较检验均采用卡方检验，检验水准α=0.05。结果 1一般情况颅内出血组167例，其中男性99例，女性68例，平均胎龄（33.59±1.95）周，平均出生体质量（1849±578）g；非颅内出血组163例，男性91例，女性72例，平均胎龄（33.98±1.63）周，平均出生体质量（1939±472）g。两组性别、出生体质量、胎龄之间差异无统计学意义，两组具有可比性。 2两组OPRMI基因A118G基因型、等位基因频率 OPRMI基因A118G位点共有两种等位基因：等位基因A、G，三种基因型：A/A型、A/G型和G/G型。颅内出血组：野生型纯合子A/A，73例(43.7%)，突变杂合子A/G，82例(49.1%)，突变纯合子G/G，12例(7.1%)；等位基因A、G频率分别为68.3%、31.7%；非颅内出血组：野生型纯合子A/A，89例(54.6%)，突变杂合子A/G，，68例(41.7%)，突变纯合子G/G，6例(3.7%)；等位基因A、G频率分别为75.5%、24.5%；两组基因型的分布没有统计学意义(χ2=4.839，P=0.089)，但是比较颅内出血组与非颅内出血组野生型(A/A)和突变型(A/G+G/G)的阳性率，二者差异有统计学意义（χ2=3.913，P=0.048），两组等位基因差异有统计学意义(χ2=4.222，P=0.04，OR=1.549，95%CI:1.003～2.391)，提示OPRMI基因118G等位基因的携带与ICH的发生呈正相关,该突变可能会增加ICH的发病风险。颅内出血组男、女性别OPRMI基因A118G多态位点基因型、等位基因比较，差异无统计学意义(χ2=0.300，P=0.58；χ2=0.843，P=0.358)；颅内出血组不同出血程度OPRMI基因A118G多态位点基因型、等位基因比较，差异无统计学意义(χ2=2.418，P=0.342；χ2=0.160，P=0.689)。结论 1. OPRMI基因A118G的单核苷酸基因多态性与早产儿颅内出血发生具有相关性。 2. OPRMI基因A118G多态性可能是ICH发病的一个潜在的易感位点，等位基因G的携带与ICH的发生呈正相关，该突变可能会增加ICH的发病风险。 3. OPRMI基因A118G单核苷酸基因多态性在颅内出血发生中无性别差异性。 4. OPRMI基因A118G单核苷酸基因多态性对颅内出血程度无影响。
[Abstract]:Background and purpose
Intracranial hemorrhage (ICH) is a kind of serious craniocerebral injury common in newborns. Preterm infants are more common, the younger the gestational age is, the higher the incidence is, the serious patients can have the sequelae of the nervous system and even death. In recent years, with the continuous development of medical technology, the survival rate of premature infants has been greatly improved, and the incidence of ICH has increased unabated.
The occurrence of ICH is related to its own anatomical and physiological characteristics and high risk factors for perinatal period. In addition, many inflammatory reactions and endogenous mediators can also be induced. The release of endogenous neuropeptides, especially endogenous opioid peptides, can aggravate this damage. In recent years, the research of neuropeptides and cerebrovascular disease mechanism has attracted more and more attention. It has been found that opioid peptide and opioid receptor system play an important role in the pathophysiological development of adult ischemic stroke.
Clinical studies have found that there are obvious individual differences in the occurrence of ICH in preterm infants with the same risk factors. The genetic background is one of the important reasons for the individual difference. The previous study found that the single nucleotide polymorphism of the platelet activating factor PAF-AH gene Val279Phe is related to the preterm infant ICH. Other susceptible genes of cord ICH.
The polymorphism of the micron opioid receptor (OPRMI) A118G gene is related to the susceptibility to a variety of diseases and the difference in the drug response. However, the study of the correlation with neonatal diseases has not yet been reported. This study is to choose the OPRMIA118G single nucleotide polymorphism, which is closely related to the mental and neurologic factors. Objective:To study the relationship between ICH and intracranial hemorrhage in premature infants and to explore the molecular genetic mechanism of ICH.
Materials and methods
1 research objects and groups
The preterm infants in the neonatal intensive care unit (NICU) of the First Affiliated Hospital of Zhengzhou University (NICU) were selected as the subjects. The intracranial hemorrhage preterm infants were treated as intracranial hemorrhage group. The non intracranial hemorrhage premature infants who were hospitalized for premature delivery were considered as non intracranial hemorrhage group, and the.2 gene polymorphism of intracranial hemorrhage was detected. Sex detection
The polymerase chain reaction restriction fragment length polymorphism analysis (Polymerase chainreaction restriction fragment length polymorphisms, PCR-RFLP) was used to detect the Al18G polymorphic loci of the OPRMI gene. The reliability.3 statistical analysis of the PCR product direct sequencing verification method of genotyping was carried out.
SPSS19.0 statistical software was used to carry out data statistics and analysis. Two groups of subjects were compared by independent sample t test, comparative test of sex composition, Hard-Weinberg balance test, comparison of genotype and allele distribution in each group, and the correlation of sex and bleeding degree with intracranial hemorrhage. Chi square test was used to test the level of alpha =0.05.
Result
1 general situation
167 cases of intracranial hemorrhage, including 99 male and 68 female, average fetal age (33.59 + 1.95) weeks, average birth weight (1849 + 578) g, non intracranial hemorrhage group 163 cases, male 91 cases, 72 cases, average gestational age (33.98 + 1.63) weeks, average birth weight (1939 + 68) g. group sex, birth body mass, birth body mass, there was no statistical difference between gestational age The group has comparability.
2 the A118G genotype and allele frequency of the two groups of OPRMI genes.
There are two alleles at the A118G locus of OPRMI gene: allele A, G, and three genotypes: A/A, A/G and G/G.
Intracerebral hemorrhage group: wild type homozygote A/A, 73 (43.7%), mutant heterozygote A/G, 82 cases (49.1%), mutant homozygote G/G, 12 cases (7.1%); allele A, G frequency 68.3%, 31.7%; non intracranial hemorrhage group: wild type homozygote A/A, 89 cases (54.6%), mutation heterozygote A/G, 68 (41.7%), mutant homozygote G/G, 6 case (3.7%); allele A, G The frequencies of the two groups were 75.5% and 24.5% respectively. The distribution of genotypes in two groups was not statistically significant (x 2=4.839, P=0.089), but the positive rates of wild type (A/A) and mutant type (A/G+G/G) in the group of intracranial hemorrhage and non intracranial hemorrhage group were statistically significant (x 2=3.913, P=0.048), and the difference between the two groups was statistically significant (x 2=4.222, P=0.04). OR=1.549,95%CI:1.003 ~ 2.391), suggesting that the 118G allele of the OPRMI gene is positively related to the occurrence of ICH. This mutation may increase the risk of ICH. The genotype of A118G polymorphic loci in the intracranial hemorrhage group, the A118G polymorphism of the female OPRMI gene, and the allele comparison of the allele, and the difference of the allele difference (x 2=0.300, P=0.58; Chi 2=0.843, P=0.358); There was no significant difference in allele genotype of OPRMI gene A118G polymorphic loci with different bleeding degree in internal hemorrhage group (x 2=2.418, P=0.342; X 2=0.160, P=0.689).
1. the single nucleotide polymorphism of OPRMI gene A118G is associated with intracranial hemorrhage in premature infants.
The 2. OPRMI gene A118G polymorphism may be a potential susceptible locus for the pathogenesis of ICH, and the carrying of the allele G is positively related to the occurrence of ICH, which may increase the risk of ICH.
3. the single nucleotide polymorphism of OPRMI A118G gene is not sex difference in intracranial hemorrhage.
4. the single nucleotide polymorphism of OPRMI A118G gene had no effect on the degree of intracranial hemorrhage.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R722.6

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