基于家庭的肿瘤坏死因子-α基因及MIF基因多态性与PCOS相关性的研究
发布时间：2018-07-16 09:00
【摘要】：研究背景多囊卵巢综合征(PCOS)是一种临床高度异质、病因复杂的疾病,但目前其发病机制仍不清楚。胰岛素抵抗(insulin resistanceIR)和高胰岛素血症是PCOS患者主要的病理生理改变,可能在PCOS的发生发展中起关键的作用。多种促炎细胞因子参与IR的发生发展,而且慢性炎症在PCOS远期并发症的发生发展中起重要作用。因此有学者提出慢性炎症可能是PCOS发病机制之一,炎症学说及其与PCOS的关系也成为研究热点。肿瘤坏死因子-α (TNF-α)可调节慢性炎症及多种代谢紊乱,在胰岛素抵抗中起重要作用。研究证实,PCOS患者血清TNF-α水平与胰岛素抵抗存在病理生理上的关系。众多研究发现PCOS患者血清TNF-α水平明显高于对照组。另有研究发现TNF-α尚参与到PCOS患者的雄激素代谢过程中,与PCOS患者的高雄激素血症相关,并且TNF-α的这种作用是独立于IR及肥胖之外的。国内外已有许多有关TNF-α基因多态性与PCOS相关性的研究,结果尚缺乏一致性。这种研究结果不一致的现象可能是由于人群分层造成的。巨噬细胞移动抑制因子(macrophage migration inhibitory factors, MIF)是一个关键的前炎症因子,是部分炎症性疾病中的关键介质,它是由T淋巴细胞分化的、具有复杂生物活性的细胞因子,具有调节巨噬细胞、淋巴细胞的免疫功能及内分泌功能的作用,也可以负反馈调节糖皮质激素的免疫抑制功能。MIF存在于各个器官组织中,参与多种疾病的病理损伤过程。有研究表明,MIF与动脉粥样硬化、冠心病、糖尿病等代谢性疾病的发生发展密切相关。MIF基因-173G/C是目前基因研究的热点,已有多项研究证实MIF-173位点与幼年特发性关节炎、溃疡性结肠炎、克隆病等自身免疫性疾病的发病有关。我中心之前的病例-对照研究发现该基因位点与PCOS发病相关。由于传统病例-对照研究设计的限制,某等位基因与某疾病的相关性可能是由因种族及环境等因素的差异引起的人群分层所导致的,也就是说病例-对照研究可能会由于可能存在的人群分层而产生假阳性结果。因此采用较传统病例-对照更准确、可靠的研究方法有助于明确PCOS的易感基因及其发病机制。以核心家系为基础的相关研究一传递不平衡检验(Transmission Disequilibrium Test,TDT),是近年来广泛采用的研究多基因病相关基因的有效方法。TDT通过评价杂合子父母的某个等位基因是否比其他等位基因优先传递给患病子女来检验该等位基因是否与某疾病相关。因为以家庭为基础的相关研究保证了病例组与对照组遗传背景的一致性,所以可以消除由于病例组与对照组的遗传背景差异而导致的假阳性关联结论。 研究目的本研究采用较传统病例-对照研究更准确、可靠的TDT进一步探讨TNF-α基因rs1799964、rs1799724及MIF基因rs755622多态性与我国PCOS发病易感性的相关性。 研究方法以216例PCOS先证者及其父亲、母亲所组成的满足经典TDT分析的核心家庭为研究对象,PCOS诊断标准按照2003年鹿特丹欧洲生殖年会(ASRM/ESHRE)修订的标准。在216个中国汉族核心家系中采用直接测序法检测TNF-α基因rs1799964、rs1799724及MIF基因rs755622多态性。PCOS患者及其父母均测定身高、体重、腰围、臀围、收缩压、舒张压,并计算体重指数、腰臀比。留取空腹血标本检测基因多态性,并测定血清甘油三酯、总胆固醇、高密度脂蛋白、低密度脂蛋白、空腹血糖、空腹胰岛素,另外PCOS患者均测定黄体生成素、卵泡刺激素、雌二醇、垂体泌乳素、睾酮水平及OGTT2小时血糖及胰岛素水平。PCOS患者根据体重指数25或≥25分为A组(非肥胖组)、B组(肥胖组),其父亲与母亲相应的根据体重指数25或≥25分为C组和D组,E组和F组。 结果216例PCOS患者的平均年龄为27.08(27.08±3.36),体重指数为24.78kg/m2(24.78±4.34)。总睾酮平均水平为58.88ng/dl(58.88±25.51)。空腹血糖及空腹胰岛素水平分别为5.63mmol/L(5.63±1.34)和12.09mIU/L(12.09±7.67)。B组与A组比较体重(P0.001)、腰围(P0.001)、臀围(P0.001)、腰臀比(P0.001)、收缩压(P0.001)、舒张压(P0.001)、甘油三酯(P=0.006)、空腹胰岛素(P0.001)、2小时胰岛素(P=0.001)均明显升高,垂体泌乳素明显降低(P=0.009),差异均具显著性。PCOS患者父亲按体重指数分为C和D组,肥胖组体重(P0.001)、腰围(P0.001)、臀围(P0.001)、腰臀比(P0.001)、收缩压(P0.001)、舒张压(P0.001)、空腹胰岛素(P0.001)、甘油三酯(P0.001)及低密度脂蛋白(P=0.025),均明显升高,两组比较均有显著性差异。F组与E组比较,身高(P=0.023)、体重(P0.001)、腰围(P0.001)、臀围(P0.001)、腰臀比(P0.001)、收缩压(P=0.005)、舒张压(P0.001)、空腹胰岛素(P0.001)及甘油三酯(P0.001)均显著升高。PCOS患者及其父母肥胖组与非肥胖组比较,rs1799964、rs1799724及rs755622基因型频率均无显著性差异。三个位点最小等位基因频率分别为0.178(rs1799964)、0.118(rs1799724)、0.191(rs755622),基因型分布均符合哈迪一温伯格平衡。在本研究的216个核心家庭中,SNP rs1799964父或母为非纯合子的家庭共有112个,rs1799724父或母为非纯合子的家庭共有76个,rs755622父或母为非纯合子的家庭共有129个,上述家庭被纳入TDT分析中。TDT提示rs1799964T (transmitted: nontransmitted=73:39; x2=10.321)、rs1799724C(transmitted: nontransmitted=43:33; x2=1.316)及rs755622C(transmitted: nontransmitted=70:59; x2=0.938)均观察到存在过度传递现象,其中rs1799964T过度传递有统计学意义(P=0.0013)。对阳性位点rs1799964进行多重检验验证,仍具有显著性差异(P=0.003)。 结论TNF-α基因rs1799964位点多态性与PCOS的发病易感性相关,T等位基因可能是增加其发病风险的关键等位基因。rs755622多态性与PCOS的发病易感性无明显相关,可能不是PCOS的易感基因。
[Abstract]:Background polycystic ovary syndrome (PCOS) is a highly heterogeneous and complicated disease, but its pathogenesis is still unclear. Insulin resistance (insulin resistanceIR) and hyperinsulinemia are the main pathophysiological changes in PCOS patients. It may play a key role in the development of PCOS. Many proinflammatory cytokines are involved in the development of PCOS. It is involved in the development of IR, and chronic inflammation plays an important role in the development of PCOS long term complications. Therefore, some scholars suggest that chronic inflammation may be one of the pathogenesis of PCOS. The theory of inflammation and its relationship with PCOS have become a hot spot. The study confirmed that the level of serum TNF- alpha in PCOS patients has a pathophysiological relationship with insulin resistance. Many studies have found that the level of TNF- alpha in serum of PCOS patients is significantly higher than that of the control group. Furthermore, the study found that TNF- alpha is still involved in the androgen metabolism of the patients with PCOS and the hormone levels of Kaohsiung in patients with PCOS. The effect of TNF- alpha is independent of IR and obesity. There are many studies on the association of TNF- alpha gene polymorphisms with PCOS at home and abroad. The results are not consistent. The inconsistent results may be caused by population stratification. The macrophage migration inhibitory factor (macrophage migration inhibito) Ry factors, MIF) is a key pro-inflammatory factor, a key medium in some inflammatory diseases. It is a cytokine that is differentiated from T lymphocytes with complex biological activity. It can regulate the immune function and endocrine function of macrophages and lymphocytes, and can also regulate the immunosuppression of glucocorticoid by negative feedback. Functional.MIF exists in various organs and tissues and participates in the pathological process of a variety of diseases. Studies have shown that MIF is closely related to the development of metabolic diseases such as atherosclerosis, coronary heart disease and diabetes, and the.MIF gene -173G/C is a hot spot in the present study. Many studies have confirmed that the MIF-173 loci and juvenile idiopathic arthritis have been confirmed. An autoimmune disease, such as ulcerative colitis, cloned disease, and other autoimmune diseases. A previous case - control study in my center found that the gene locus was associated with PCOS. The correlation of a certain allele to a certain disease may be caused by differences in species and environment due to the restriction of the traditional case control study design. It is said that case control studies may result in false positive results due to the stratification of possible populations. Therefore, it is more accurate and reliable to use a more accurate and reliable method to identify the susceptibility genes and pathogenesis of PCOS. Nsmission Disequilibrium Test, TDT), which is widely used in recent years to study the genes related to polygenous diseases,.TDT by evaluating whether a allele of heterozygotes is preferentially transferred to a sick child to determine whether the allele is associated with a certain disease. The study ensures the consistency of the genetic background of the case group and the control group, so it can eliminate the false positive association results due to the genetic background difference between the case group and the control group.
The purpose of this study was to further explore the correlation between the polymorphisms of the TNF- alpha gene rs1799964, rs1799724 and MIF gene and the susceptibility to PCOS in China by using a more accurate and reliable TDT than the traditional case control study.
The study was based on the core families of 216 PCOS precursor and their fathers and their fathers and mothers to meet the classic TDT analysis. The PCOS diagnostic standard was based on the revised standard of the 2003 Rotterdam European reproductive year (ASRM/ESHRE). The direct sequencing method was used to detect the TNF- alpha gene rs1799964, rs1799724 and M in 216 Chinese Han core families. IF gene rs755622 polymorphism.PCOS patients and their parents all measured height, weight, waist circumference, systolic pressure, diastolic pressure, and calculated body mass index, waist to hip ratio, left fasting blood specimen to detect gene polymorphism, and determination of serum triglyceride, total cholesterol, high density lipoprotein, low density lipoprotein, fasting glucose, fasting insulin, and PCOS The patients were measured with luteinizing hormone, follicular stimulating hormone, estradiol, pituitary prolactin, testosterone level, OGTT2 hour blood glucose and insulin level in.PCOS patients, according to body mass index 25 or more than 25 (non obese group), group B (obese group), their father and mother's corresponding body mass index 25 or 25 scores were C and D group, E group and F group.
Results the average age of 216 patients with PCOS was 27.08 (27.08 + 3.36), the body mass index was 24.78kg/m2 (24.78 + 4.34). The average level of total testosterone was 58.88ng/dl (58.88 + 25.51). The level of fasting blood glucose and fasting insulin was 5.63mmol/L (5.63 + 1.34) and 12.09mIU/L (12.09 + 7.67).B group with A group (P0.001), waist circumference (P0.001), hip circumference (P0.0). 01), the waist to hip ratio (P0.001), the systolic pressure (P0.001), the diastolic pressure (P0.001), the triglyceride (P=0.006), the fasting insulin (P0.001), the 2 hour insulin (P=0.001) were all significantly increased, and the pituitary prolactin was significantly decreased (P=0.009). The differences were all marked by the body mass index in the group of C and D, the weight of the obese group (P0.001), the waist circumference (P0.001), and the hip circumference. 001) the waist to hip ratio (P0.001), systolic pressure (P0.001), diastolic pressure (P0.001), fasting insulin (P0.001), triglyceride (P0.001) and low density lipoprotein (P=0.025) were all significantly increased. The comparison between the two groups was significantly different from that in the E group, the height (P= 0.023), the weight (P0.001), the waist circumference (P0.001), the hip circumference (P0.001), the waist to hip ratio, systolic pressure. Diastolic pressure (P0.001), fasting insulin (P0.001) and triglyceride (P0.001) increased significantly in.PCOS patients and their parents and their obese and non obese groups. There was no significant difference in the frequencies of rs1799964, rs1799724 and rs755622 genotypes. The minimum allele frequency of three loci was 0.178 (rs1799964), 0.118 (rs1799724), 0.191 (rs755622), respectively. Among the 216 core families of this study, there are 112 families with SNP rs1799964 father or mother as non homozygous families, 76 families with rs1799724 father or mother as non homozygous families, 129 families with non homozygous parents, and 129 families with non homozygous parents, and these families are included in TDT analysis and.TDT prompts rs1799. 964T (transmitted: nontransmitted=73:39; x2=10.321), rs1799724C (transmitted: nontransmitted=43:33; x2=1.316) and rs755622C (transmitted: nontransmitted=70:59; x2=0.938) observed the existence of excessive transmission. There are still significant differences (P=0.003).
Conclusion the polymorphism of the TNF- alpha gene rs1799964 locus is associated with the susceptibility to PCOS. The T allele may not be the key allele of the key allele to increase the risk of the disease. There is no significant correlation between the polymorphism of the allele.Rs755622 and the susceptibility to PCOS. It may not be a susceptible gene of PCOS.
【学位授予单位】：山东大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R711.75
[Abstract]:Background polycystic ovary syndrome (PCOS) is a highly heterogeneous and complicated disease, but its pathogenesis is still unclear. Insulin resistance (insulin resistanceIR) and hyperinsulinemia are the main pathophysiological changes in PCOS patients. It may play a key role in the development of PCOS. Many proinflammatory cytokines are involved in the development of PCOS. It is involved in the development of IR, and chronic inflammation plays an important role in the development of PCOS long term complications. Therefore, some scholars suggest that chronic inflammation may be one of the pathogenesis of PCOS. The theory of inflammation and its relationship with PCOS have become a hot spot. The study confirmed that the level of serum TNF- alpha in PCOS patients has a pathophysiological relationship with insulin resistance. Many studies have found that the level of TNF- alpha in serum of PCOS patients is significantly higher than that of the control group. Furthermore, the study found that TNF- alpha is still involved in the androgen metabolism of the patients with PCOS and the hormone levels of Kaohsiung in patients with PCOS. The effect of TNF- alpha is independent of IR and obesity. There are many studies on the association of TNF- alpha gene polymorphisms with PCOS at home and abroad. The results are not consistent. The inconsistent results may be caused by population stratification. The macrophage migration inhibitory factor (macrophage migration inhibito) Ry factors, MIF) is a key pro-inflammatory factor, a key medium in some inflammatory diseases. It is a cytokine that is differentiated from T lymphocytes with complex biological activity. It can regulate the immune function and endocrine function of macrophages and lymphocytes, and can also regulate the immunosuppression of glucocorticoid by negative feedback. Functional.MIF exists in various organs and tissues and participates in the pathological process of a variety of diseases. Studies have shown that MIF is closely related to the development of metabolic diseases such as atherosclerosis, coronary heart disease and diabetes, and the.MIF gene -173G/C is a hot spot in the present study. Many studies have confirmed that the MIF-173 loci and juvenile idiopathic arthritis have been confirmed. An autoimmune disease, such as ulcerative colitis, cloned disease, and other autoimmune diseases. A previous case - control study in my center found that the gene locus was associated with PCOS. The correlation of a certain allele to a certain disease may be caused by differences in species and environment due to the restriction of the traditional case control study design. It is said that case control studies may result in false positive results due to the stratification of possible populations. Therefore, it is more accurate and reliable to use a more accurate and reliable method to identify the susceptibility genes and pathogenesis of PCOS. Nsmission Disequilibrium Test, TDT), which is widely used in recent years to study the genes related to polygenous diseases,.TDT by evaluating whether a allele of heterozygotes is preferentially transferred to a sick child to determine whether the allele is associated with a certain disease. The study ensures the consistency of the genetic background of the case group and the control group, so it can eliminate the false positive association results due to the genetic background difference between the case group and the control group.
The purpose of this study was to further explore the correlation between the polymorphisms of the TNF- alpha gene rs1799964, rs1799724 and MIF gene and the susceptibility to PCOS in China by using a more accurate and reliable TDT than the traditional case control study.
The study was based on the core families of 216 PCOS precursor and their fathers and their fathers and mothers to meet the classic TDT analysis. The PCOS diagnostic standard was based on the revised standard of the 2003 Rotterdam European reproductive year (ASRM/ESHRE). The direct sequencing method was used to detect the TNF- alpha gene rs1799964, rs1799724 and M in 216 Chinese Han core families. IF gene rs755622 polymorphism.PCOS patients and their parents all measured height, weight, waist circumference, systolic pressure, diastolic pressure, and calculated body mass index, waist to hip ratio, left fasting blood specimen to detect gene polymorphism, and determination of serum triglyceride, total cholesterol, high density lipoprotein, low density lipoprotein, fasting glucose, fasting insulin, and PCOS The patients were measured with luteinizing hormone, follicular stimulating hormone, estradiol, pituitary prolactin, testosterone level, OGTT2 hour blood glucose and insulin level in.PCOS patients, according to body mass index 25 or more than 25 (non obese group), group B (obese group), their father and mother's corresponding body mass index 25 or 25 scores were C and D group, E group and F group.
Results the average age of 216 patients with PCOS was 27.08 (27.08 + 3.36), the body mass index was 24.78kg/m2 (24.78 + 4.34). The average level of total testosterone was 58.88ng/dl (58.88 + 25.51). The level of fasting blood glucose and fasting insulin was 5.63mmol/L (5.63 + 1.34) and 12.09mIU/L (12.09 + 7.67).B group with A group (P0.001), waist circumference (P0.001), hip circumference (P0.0). 01), the waist to hip ratio (P0.001), the systolic pressure (P0.001), the diastolic pressure (P0.001), the triglyceride (P=0.006), the fasting insulin (P0.001), the 2 hour insulin (P=0.001) were all significantly increased, and the pituitary prolactin was significantly decreased (P=0.009). The differences were all marked by the body mass index in the group of C and D, the weight of the obese group (P0.001), the waist circumference (P0.001), and the hip circumference. 001) the waist to hip ratio (P0.001), systolic pressure (P0.001), diastolic pressure (P0.001), fasting insulin (P0.001), triglyceride (P0.001) and low density lipoprotein (P=0.025) were all significantly increased. The comparison between the two groups was significantly different from that in the E group, the height (P= 0.023), the weight (P0.001), the waist circumference (P0.001), the hip circumference (P0.001), the waist to hip ratio, systolic pressure. Diastolic pressure (P0.001), fasting insulin (P0.001) and triglyceride (P0.001) increased significantly in.PCOS patients and their parents and their obese and non obese groups. There was no significant difference in the frequencies of rs1799964, rs1799724 and rs755622 genotypes. The minimum allele frequency of three loci was 0.178 (rs1799964), 0.118 (rs1799724), 0.191 (rs755622), respectively. Among the 216 core families of this study, there are 112 families with SNP rs1799964 father or mother as non homozygous families, 76 families with rs1799724 father or mother as non homozygous families, 129 families with non homozygous parents, and 129 families with non homozygous parents, and these families are included in TDT analysis and.TDT prompts rs1799. 964T (transmitted: nontransmitted=73:39; x2=10.321), rs1799724C (transmitted: nontransmitted=43:33; x2=1.316) and rs755622C (transmitted: nontransmitted=70:59; x2=0.938) observed the existence of excessive transmission. There are still significant differences (P=0.003).
Conclusion the polymorphism of the TNF- alpha gene rs1799964 locus is associated with the susceptibility to PCOS. The T allele may not be the key allele of the key allele to increase the risk of the disease. There is no significant correlation between the polymorphism of the allele.Rs755622 and the susceptibility to PCOS. It may not be a susceptible gene of PCOS.
【学位授予单位】：山东大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R711.75
【参考文献】
相关期刊论文 前10条
1 马元涛;梁秀利;;慢性丙肝患者血清瘦素、肿瘤坏死因子α含量与体内胰岛素水平的关系[J];中国医师杂志;2006年04期
2 张琴芬;韩素萍;崔毓桂;李瑛;马翔;高莉;冒韵东;刘嘉茵;;胰岛素基因可变重复序列多态性与PCOS的相关性[J];国际生殖健康/计划生育杂志;2010年04期
3 张新林;张成伟;王勇;;多囊卵巢综合征的基因多态性研究进展[J];国际妇产科学杂志;2012年02期
4 李游;范平;白怀;万德花;刘宏伟;;PCOS患者PAF-AH活性及其与糖脂代谢相关性的研究[J];四川大学学报(医学版);2009年06期
5 李蓉;许良智;熊冬梅;;Calpain-10基因多态性与PCOS患者遗传易感性的相关性研究[J];四川大学学报(医学版);2010年02期
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