基于家庭的腫瘤壞死因子-α基因及MIF基因多態(tài)性與PCOS相關性的研究
發(fā)布時間：2018-07-16 09:00
【摘要】：研究背景多囊卵巢綜合征(PCOS)是一種臨床高度異質、病因復雜的疾病,但目前其發(fā)病機制仍不清楚。胰島素抵抗(insulin resistanceIR)和高胰島素血癥是PCOS患者主要的病理生理改變,可能在PCOS的發(fā)生發(fā)展中起關鍵的作用。多種促炎細胞因子參與IR的發(fā)生發(fā)展,而且慢性炎癥在PCOS遠期并發(fā)癥的發(fā)生發(fā)展中起重要作用。因此有學者提出慢性炎癥可能是PCOS發(fā)病機制之一,炎癥學說及其與PCOS的關系也成為研究熱點。腫瘤壞死因子-α (TNF-α)可調節(jié)慢性炎癥及多種代謝紊亂,在胰島素抵抗中起重要作用。研究證實,PCOS患者血清TNF-α水平與胰島素抵抗存在病理生理上的關系。眾多研究發(fā)現PCOS患者血清TNF-α水平明顯高于對照組。另有研究發(fā)現TNF-α尚參與到PCOS患者的雄激素代謝過程中,與PCOS患者的高雄激素血癥相關,并且TNF-α的這種作用是獨立于IR及肥胖之外的。國內外已有許多有關TNF-α基因多態(tài)性與PCOS相關性的研究,結果尚缺乏一致性。這種研究結果不一致的現象可能是由于人群分層造成的。巨噬細胞移動抑制因子(macrophage migration inhibitory factors, MIF)是一個關鍵的前炎癥因子,是部分炎癥性疾病中的關鍵介質,它是由T淋巴細胞分化的、具有復雜生物活性的細胞因子,具有調節(jié)巨噬細胞、淋巴細胞的免疫功能及內分泌功能的作用,也可以負反饋調節(jié)糖皮質激素的免疫抑制功能。MIF存在于各個器官組織中,參與多種疾病的病理損傷過程。有研究表明,MIF與動脈粥樣硬化、冠心病、糖尿病等代謝性疾病的發(fā)生發(fā)展密切相關。MIF基因-173G/C是目前基因研究的熱點,已有多項研究證實MIF-173位點與幼年特發(fā)性關節(jié)炎、潰瘍性結腸炎、克隆病等自身免疫性疾病的發(fā)病有關。我中心之前的病例-對照研究發(fā)現該基因位點與PCOS發(fā)病相關。由于傳統(tǒng)病例-對照研究設計的限制,某等位基因與某疾病的相關性可能是由因種族及環(huán)境等因素的差異引起的人群分層所導致的,也就是說病例-對照研究可能會由于可能存在的人群分層而產生假陽性結果。因此采用較傳統(tǒng)病例-對照更準確、可靠的研究方法有助于明確PCOS的易感基因及其發(fā)病機制。以核心家系為基礎的相關研究一傳遞不平衡檢驗(Transmission Disequilibrium Test,TDT),是近年來廣泛采用的研究多基因病相關基因的有效方法。TDT通過評價雜合子父母的某個等位基因是否比其他等位基因優(yōu)先傳遞給患病子女來檢驗該等位基因是否與某疾病相關。因為以家庭為基礎的相關研究保證了病例組與對照組遺傳背景的一致性,所以可以消除由于病例組與對照組的遺傳背景差異而導致的假陽性關聯(lián)結論。 研究目的本研究采用較傳統(tǒng)病例-對照研究更準確、可靠的TDT進一步探討TNF-α基因rs1799964、rs1799724及MIF基因rs755622多態(tài)性與我國PCOS發(fā)病易感性的相關性。 研究方法以216例PCOS先證者及其父親、母親所組成的滿足經典TDT分析的核心家庭為研究對象,PCOS診斷標準按照2003年鹿特丹歐洲生殖年會(ASRM/ESHRE)修訂的標準。在216個中國漢族核心家系中采用直接測序法檢測TNF-α基因rs1799964、rs1799724及MIF基因rs755622多態(tài)性。PCOS患者及其父母均測定身高、體重、腰圍、臀圍、收縮壓、舒張壓,并計算體重指數、腰臀比。留取空腹血標本檢測基因多態(tài)性,并測定血清甘油三酯、總膽固醇、高密度脂蛋白、低密度脂蛋白、空腹血糖、空腹胰島素,另外PCOS患者均測定黃體生成素、卵泡刺激素、雌二醇、垂體泌乳素、睪酮水平及OGTT2小時血糖及胰島素水平。PCOS患者根據體重指數25或≥25分為A組(非肥胖組)、B組(肥胖組),其父親與母親相應的根據體重指數25或≥25分為C組和D組,E組和F組。 結果216例PCOS患者的平均年齡為27.08(27.08±3.36),體重指數為24.78kg/m2(24.78±4.34)�？偛G酮平均水平為58.88ng/dl(58.88±25.51)�？崭寡羌翱崭挂葝u素水平分別為5.63mmol/L(5.63±1.34)和12.09mIU/L(12.09±7.67)。B組與A組比較體重(P0.001)、腰圍(P0.001)、臀圍(P0.001)、腰臀比(P0.001)、收縮壓(P0.001)、舒張壓(P0.001)、甘油三酯(P=0.006)、空腹胰島素(P0.001)、2小時胰島素(P=0.001)均明顯升高,垂體泌乳素明顯降低(P=0.009),差異均具顯著性。PCOS患者父親按體重指數分為C和D組,肥胖組體重(P0.001)、腰圍(P0.001)、臀圍(P0.001)、腰臀比(P0.001)、收縮壓(P0.001)、舒張壓(P0.001)、空腹胰島素(P0.001)、甘油三酯(P0.001)及低密度脂蛋白(P=0.025),均明顯升高,兩組比較均有顯著性差異。F組與E組比較,身高(P=0.023)、體重(P0.001)、腰圍(P0.001)、臀圍(P0.001)、腰臀比(P0.001)、收縮壓(P=0.005)、舒張壓(P0.001)、空腹胰島素(P0.001)及甘油三酯(P0.001)均顯著升高。PCOS患者及其父母肥胖組與非肥胖組比較,rs1799964、rs1799724及rs755622基因型頻率均無顯著性差異。三個位點最小等位基因頻率分別為0.178(rs1799964)、0.118(rs1799724)、0.191(rs755622),基因型分布均符合哈迪一溫伯格平衡。在本研究的216個核心家庭中,SNP rs1799964父或母為非純合子的家庭共有112個,rs1799724父或母為非純合子的家庭共有76個,rs755622父或母為非純合子的家庭共有129個,上述家庭被納入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)均觀察到存在過度傳遞現象,其中rs1799964T過度傳遞有統(tǒng)計學意義(P=0.0013)。對陽性位點rs1799964進行多重檢驗驗證,仍具有顯著性差異(P=0.003)。 結論TNF-α基因rs1799964位點多態(tài)性與PCOS的發(fā)病易感性相關,T等位基因可能是增加其發(fā)病風險的關鍵等位基因。rs755622多態(tài)性與PCOS的發(fā)病易感性無明顯相關,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.
【學位授予單位】：山東大學
【學位級別】：博士
【學位授予年份】：2014
【分類號】：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.
【學位授予單位】：山東大學
【學位級別】：博士
【學位授予年份】：2014
【分類號】：R711.75
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