TGF-β1基因多态性与广东地区子痫前期相关性研究

目的:探讨转化生长因子β1(TGF-β1)基因4个位点-800G/A、-509C/T、+869T/C、+915G/C的单核苷酸多态性(SNP)及其构成的单体型与子痫前期(PE)的相关性。方法:随机选取2015年1月至12月于广州医科大学附属第三医院住院的广东地区汉族孕妇176例,其中PE患者88例(PE组)、正常妊娠88例(对照组)。利用SNa Pshot技术对TGF-β1基因4个位点进行基因分型检测,分析PE组与对照组的TGF-β1 4个SNP基因型分布及等位基因频率的差异,并对+869T/C、-509C/T两个位点构建单体型分析。结果:(1)中国广东地区汉族妇女中存在TGF-β1+869T/C、-509C/T多态性,暂未发现-800G/A、+915G/C多态性。(2)PE组+869T/C位点CC基因型、C等位基因频率明显高于对照组(44.32%vs 27.27%,P=0.028,OR=1.935,95%CI为1.028~3.639;68.18%vs 56.25%,P=0.014,OR=1.667,95%CI为1.079~2.575),TT基因型和T等位基因则明显下降(5.68%vs 14.77%,P=0.040,OR=0.348,95%CI为0.118~1.021;31.82%vs 43.75%,P=0.014,OR=0.600,95%CI为0.388~0.927);两组CT基因型分布无明显差异(52.27%vs 57.95%,P=0.272)。(3)PE与对照组比较,-509C/T位点CC、CT、TT基因型分布无明显差异(14.77%vs 23.86%,P=0.090;47.73%vs 50.00%,P=0.440;37.50%vs 26.14%,P=0.072),而C等位基因明显下降(38.64%vs 48.86%,P=0.034,OR=0.659,95%CI为0.431~1.006),T等位基因明显升高(61.36%vs51.14%,P=0.034,OR=1.518,95%CI为0.994~2.318)。(4)单体型分析发现,由+869T/C、-509C/T两个位点构建的C-T单体型在PE组明显增加(62.79%vs 49.43%,P=0.008,OR=1.727,95%CI为1.125~2.651),而T-C单体型明显下降(31.40%vs42.33%,P=0.021,OR=0.618,95%CI为0.398~0.961)。结论:(1)TGF-β1基因+869T/C、-509C/T SNPs可能与中国广东地区汉族妇女PE发生有关,而暂未发现-800G/A、+915G/C多态性。(2)TGF-β1基因SNP+869T/CCC基因型、C等位基因,-509C/TT等位基因可能是广东汉族妇女PE的危险因素,而+869T/CTT基因型、T等位基因,-509C/TC等位基因可能是保护因素。(3)2个多态位点构成的C-T单体型可能是中国广东地区汉族妇女PE的危险因素,T-C单体型则是保护因素。     

转化生长因子β1基因多态性与广东地区汉族妊娠期糖尿病的相关性研究

目的:探讨转化生长因子β1(TGF-β1)4个基因位点:-800G/A、-509C/T、+869T/C、+915G/C的单核苷酸多态性(SNP)及其构成的单体型与中国广东地区汉族女性妊娠期糖尿病(GDM)发生风险的相关性。方法:回顾性分析2014年9月至2017年3月在广州医科大学附属第三医院产科产前检查并分娩的广东地区汉族女性240例(正常对照组120例,GDM组120例)。利用SNaPshot基因测序技术对各组TGF-β1的4个基因位点(-800G/A、-509C/T、+869T/C、+915G/C)进行基因分型检测,并对各个位点的基因型频率、等位基因频率及其构成的单体型与GDM的相关性进行比较分析。结果:①GDM组与正常对照组TGF-β1的+915 G/C、-800G/A 2个基因位点暂未发现SNP多态性,而-509C/T、+869T/C 2个基因位点存在多态性。②GDM组+869T/C位点的CC基因型、C等位基因频率明显高于正常对照组(55.83%vs 32.50%、75.42%vs 54.58%),而TT基因型和T等位基因明显低于正常对照组(5.00%vs 23.33%、24...     

NBS1基因多态与宫颈癌发生的相关性研究

目的:分析DNA双链损伤修复基因NBS1的Glu185Gln单核苷酸多态性与广东地区汉族妇女的宫颈癌遗传易感性的关系。方法:采用聚合酶链式反应-限制性片段长度多态性(PCR-RFLP)技术,对139例宫颈癌患者及264例健康对照者的NBS1Glu185Gln多态位点进行基因分型,比较病例组与对照组的基因型分布情况,并采用非条件logistic回归方法分析该多态位点与宫颈癌发生的关系。结果:携带GG基因型的个体患宫颈癌的风险显著增加(OR=2.231,95%CI=1.247~3.991,P=0.007),并且G等位基因增加个体患宫颈癌风险的趋势在Ⅰ期的宫颈癌患者中更显著(GG vs CC:调整后OR=3.703,95%CI=1.559~8.798,P=0.003)。结论:NBS1基因的Glu185Gln多态位点与广东地区汉族妇女宫颈癌的发生相关,G等位基因可能是影响宫颈癌易感的风险因素。     

TBX1基因多态性与先天性心脏病的相关性

目的探讨TBX1基因多态性与先天性心脏病(先心病)的相关关系。 方法选择2004-10—2005-03在哈尔滨医科大学附属第二医院住院的先心病患儿99例，对照组96名(与先心病患儿年龄相近的健康儿童)。应用小剂量DNA提取技术及采用ABI Prism SNaPshot方法,检测了先心病患儿和对照组儿童的TBX1基因多态性。 结果经χ2检验，TBX1基因各基因型频率在对照组、先心病组之间均无显著性差异(P>005)；各等位基因频率在两组人群中差异均无显著性意义(P>005)。 结论未发现TBX1基因多态性与先天性心脏病之间存在相关关系。     

脂联素基因多态性与妊娠期糖尿病的相关性研究

目的:探讨脂联素基因启动子区单核苷酸多态性与妊娠期糖尿病(GDM)的相关性。方法:采用基质辅助激光解吸电离飞行时间质谱(MALDI-TOFMS)技术,检测103例GDM孕妇和97例正常孕妇脂联素基因启动子区-11426A>G及-11377C>G多态位点,并测定40例未经任何治疗的GDM孕妇FINS、FPG、TC、TG、HDL-C、LDL-C、脂联素水平,同时计算稳态模型的胰岛素抵抗指数。结果:(1)GDM孕妇脂联素基因-11426A>G位点的AG、GG基因型频率显著高于对照组(χ2=8.822,P=0.012),G等位基因频率明显高于对照组(χ2=10.283,P=0.001)。-11377C>G位点基因型频率及等位基因频率在二组孕妇中分布无显著差异(P>0.05)。(2)GDM孕妇脂联素-11426A>G多态性位点AG+GG基因型组甘油三酯水平高于AA基因型组(P=0.023),血清脂联素水平低于AA基因型组(P=0.024)。结论:脂联素基因-11426A>G位点多态性可能与GDM的发生有关,G等位基因在GDM孕妇中分布频率增加,A→G突变可能通过降低血清脂联素水平参与GDM的发生。     

胎盘中传递甲状腺激素的转运蛋白研究

甲状腺激素对机体各组织的新陈代谢起着重要的作用,也是人类妊娠期重要的内分泌激素。研究证实,在胎儿自身甲状腺激素 分泌前,母体甲状腺激素能通过胎盘并参与胎儿内分泌系统的调节。这个过程中需要一些特殊的转运蛋白,有关人胎盘中的这些重要的转运蛋白 相关研究很少,已证实的有人类胎盘中氨基酸转运载体、有机阴离子转运肽、单羧酸转运肽、Na+/I-同向转运蛋白等。但仍需进一步研究胎盘中 转运甲状腺激素的一些蛋白作用,以了解导致胎儿生长受限、流产、胚芽种植失败、胎儿死亡等不良妊娠结局的发生机制。     

ABCA1基因R219K与血脂及子痫前期易感性的相关性研究

目的探讨福建省汉族人群三磷腺苷结合转运子A1基因（ABCA1）R219K单核苷酸多态性（SNP）和子痫前期（preeclampsia）及其血脂水平的关联性。方法研究对象621例,包括对照组316例,子痫前期病例组305例。采用聚合酶链反应一限制性片段长度多态性（PCR-RFLP）法鉴定ABCA1基因外显子区R219K单核苷酸多态性并测定血脂水平。结果 ABCA1基因R219K位点K等位基因频率和RK＋KK基因型频率在子痫前期病例组明显降低,差异有高度统计学意义P〈0.001）。子痫前期病例组内RK＋KK基因型患者血清甘油三酯（TG）浓度低于RR基因型,而HDL-C浓度刚好相反,差异有高度统计学意义（P〈0.001）。结论福建省汉族人群ABCA1基因R219K中,K等位基因可能是子痫前期的独立的保护因子,其机制可能是通过提高血液中HDL-C水平,降低TG水平,从而降低了子痫前期合并血脂代谢紊乱的风险性。     

胎盘中传递甲状腺激素的转运蛋白研究

甲状腺激素对机体各组织的新陈代谢起着重要的作用,也是人类妊娠期重要的内分泌激素。研究证实,在胎儿自身甲状腺激素分泌前,母体甲状腺激素能通过胎盘并参与胎儿内分泌系统的调节。这个过程中需要一些特殊的转运蛋白,有关人胎盘中的这些重要的转运蛋白相关研究很少,已证实的有人类胎盘中氨基酸转运载体、有机阴离子转运肽、单羧酸转运肽、Na^＋/I^-同向转运蛋白等。但仍需进一步研究胎盘中转运甲状腺激素的一些蛋白作用,以了解导致胎儿生长受限、流产、胚芽种植失败、胎儿死亡等不良妊娠结局的发生机制。     

TGF-β1及其基因多态性与妊娠期糖尿病的相关性研究进展

妊娠期糖尿病(GDM)发病机制未明,其病理生理基础为胰岛素抵抗及胰岛β细胞分泌缺陷。转化生长因子-β1(TGF-β1)是一类含多种生物活性的多肽生长因子,具有调节细胞增殖与分化、胚胎发育、血管生成、免疫抑制等功能。国内外有研究发现,TGF-β1表达异常与GDM发生相关,而其基因多态性影响其转录和翻译,可能与GDM的发生、发展密切相关。本文就近年国内外TGF-β1基因多态性与GDM发病机制的相关性的研究进展做一综述。     

胆固醇酯转运蛋白基因多态性在多囊卵巢综合征中的初步研究

目的:探讨胆固醇酯转运蛋白(CETP)在PCOS患者中的基因多态性和血脂、性激素的关系,以进一步认识PCOS脂代谢的特点。方法:应用PCR酶切方法检测108例PCOS患者及60例对照组的CETPTaqIB位点基因多态性。结果:CETPTaqIB基因频率及基因型分布在两组间分布差异无显著性。结论:CETPTaqIB基因多态性影响PCOS患者的HDLC水平,该基因在PCOS患者分布与正常人群分布无差异性。     

Expression of the multidrug resistance P-glycoprotein, (ABCB1 glycoprotein) in the human placenta decreases with advancing gestation

The multidrug resistance p-glycoprotein (P-gp), encoded by the ABCB1 gene, is a plasma membrane protein that actively extrudes a wide variety of substances from cells. Preliminary studies in mice have shown that the ABCB1/P-gp can protect the fetus from a number of toxic substances. ABCB1/P-gp is expressed in the human placenta and is potentially capable of protecting the fetus from a large number of drugs and toxins, including herbicides and pesticides. The protein can also extrude various steroids including certain glucocorticoids and may therefore play an important role in regulating fetal access of glucocorticoids. The aim of the present study was to examine the expression profile and cellular localization of ABCB1/P-gp in human placenta throughout gestation. We hypothesized that there would be gestational age-related changes in the expression of the protein. ABCB1/P-gp mRNA was measured by Real-Time PCR using specific probes in tissues obtained from 6 weeks gestation to term. ABCB1/P-gp mRNA levels in placental tissue obtained at 6-10 weeks (n=5) and 24-35 weeks (n=5) were significantly higher than in tissues obtained at term (38-41 weeks gestation) by elective C-section (n=6) or following labor (n=6). The profile of ABCB1/P-gp protein levels, quantified using Western analysis, demonstrated a similar decrease with advancing gestation. At all gestational ages ABCB1/P-gp was localized by immunohistochemistry to the syncytiotrophoblast. In term tissues, it appeared to be localized to some areas of the villi and not others. Together, these data indicate that with advancing gestation there is a decrease in the level of ABCB1/P-gp in the human placenta indicating that the fetus may be more susceptible to toxic insults in the latter part of gestation. Further, the reduction in ABCB1/P-gp expression may contribute to the increased transfer of maternal cortisol to the fetus that is known to occur in late gestation.     

Expression, localisation and activity of ATP binding cassette (ABC) family of drug transporters in human amnion membranes

Placental ATP-binding cassette (ABC) transporters limit fetal exposure to xenobiotics by regulating transplacental passage into the fetal circulation; their expression and function in fetal membranes, however, has not been studied. In the present study the expression, localisation and function of ABC transporters in human amnion was examined to explore their potential role in modulating amniotic fluid drug disposition in pregnancy. Single-assay oligo-microarrays were used to profile amnion gene expression, and drug transporters expressed at significant levels were identified and selected for further studies. The expression of ABCG2/breast cancer resistance protein (BCRP) and multidrug resistance-associated proteins (MRP) 1 (ABCC1), 2 (ABCC2) and 5 (ABCC5) was detected on the arrays, and verified by RT-PCR and immunoblotting. On confocal microscopy of fetal membrane cryosections, MRP1 and MRP5 were immunolocalised to both apical and basolateral surfaces of the amniotic epithelium, while MRP2 was expressed at low levels only in the apical membrane. BCRP in contrast showed cytoplasmic staining throughout the amniotic epithelium. In addition to the amnion, MRP1 and BCRP immunostaining was observed in the chorion and the decidua. Cell accumulation studies using selective MRP and BCRP inhibitors showed the transporters to be functionally active in amnion epithelial monolayer cultures. In contrast, transwell transport studies using intact amnion membranes did not show significant vectorial transport. These findings identify the amnion as a novel site of ABC drug transporter expression. Functional studies indicate that they may act primarily to prevent cellular xenobiotic accumulation, rather than to confer fetal protection through reduced accumulation in amniotic fluid.     

MTNR1A and MTNR1B gene polymorphisms in women with gestational diabetes

Gestational diabetes mellitus (GDM) is glucose intolerance detected during pregnancy. The MTNR1B gene is the genetic locus associated with type 2 diabetes, that may affect insulin secretion and pancreatic glucose sensing. In this study, we examined the association between MTNR1A (rs2119882) and MTNR1B (rs10830963, rs4753426) gene polymorphisms and the risk of GDM. According to the results of their oral glucose tolerance test (OGTT), the women were divided into two groups: 204 pregnant women with GDM and 207 pregnant women with normal glucose tolerance (NGT). There were no statistically significant differences in the distribution of MTNR1A rs2119882 and MTNR1B rs4753426 genotypes and alleles between women with GDM and healthy pregnant women. With regard to the MTNR1B rs10830963 polymorphism, we observed a statistically significant prevalence of GG and CG genotypes and the G allele among pregnant women with GDM (GG?+?CG vs CC, OR 1.50, 95% CI 1.02–2.22, p?=?0.04; G vs C, OR 1.43, 95% CI 1.07–1.90, p?=?0.016). In a multivariate logistic regression analysis, a higher number of MTNR1B rs10830963?G alleles was an independent significant predictor of a higher risk of GDM. The results of our study indicate that MTNR1B rs10830963 polymorphism is associated with GDM susceptibility, and women with a higher number of G alleles have an increased risk of GDM development.     

Ultracytochemical localization of NAD(P)H oxidase activity in the human placenta

Ultracytochemical localization of NAD(P)H oxidase activity was demonstrated in the human term placenta by the cerium method. The activity of this enzyme was also compared to those of other oxygen-intermediates-metabolizing enzymes, such as xanthine oxidase, catalase, peroxidase and superoxide dismutase. NAD(P)H oxidase activity was exclusively confined to the apical microvillous membrane of the syncytiotrophoblast. Other enzymes studied showed no activity. We discuss the possibility that NAD(P)H oxidase might play a role in transferring substances between mother and fetus and that this enzyme might modulate placental H2O2 production.     

Presence of gamma-aminobutyric acid and its specific receptor binding sites in the human term placenta

The concentration of gamma-aminobutyric acid (GABA), the activity of glutamate decarboxylase (GAD), and the presence of specific GABA receptor binding sites have been examined in human term placentas. The estimated values of GABA concentration and GAD activity in the placenta were about 50 nmol/g tissue and 1.1 nmol/mg protein/h, respectively. A remarkable density of high-affinity and specific GABA binding sites has also been demonstrated in membranes of the human term placenta. These binding sites showed the properties of a GABAA receptor. The present findings suggest a possible role of GABA in the function of human placenta.     

Lack of association between the TGF-beta1 gene polymorphisms and recurrent spontaneous abortion

Transforming growth factor-beta1 (TGF-beta1) is produced by T regulatory lymphocytes (Treg), which play an important role in the physiology of pregnancy. Several polymorphisms of the TGF-beta1 gene (TGFB1) have been reported, some with an important correlation with TGF-beta1 production and disease severity. We performed an association study between TGFB1 polymorphisms and recurrent spontaneous abortion (RSA). We first used a PCR-RFLP method to detect three known TGFB1 cSNPs (coding single nucleotide polymorphisms) among 111 RSA and 110 normal control women from Southern Iran, such as 29T-->C (Leu 10 Pro), 74G-->C (Arg 25 Pro) and 788C-->T (Thr 263Ile), and compared their frequencies between the two groups of subjects. To confirm results of the RFLP study and to identify new SNPs in the RSA women, we then sequenced their DNA samples for seven exons and adjacent intronic regions of TGFB1. Consequently, 10 SNPs were detected; one (-14G-->A) was located in the upstream region of exon 1, three in exons (two in exon 1 and one in exon 5) and six in intronic regions. Two (IVS5+18G-->C and IVS6+910G-->A) of the 10 SNPs were novel. Statistical analysis on the frequency of six most frequent SNPs, including the three cSNPs, as well as on the frequencies of genotypes and 13 haplotypes regarding the 6 SNPs, revealed no significant difference between RSA and control women. Therefore, this study concludes that there is no association between exonic and adjacent intronic polymorphisms of TGFB1 and RSA.     

Hepatocyte growth factor activator (HGF-A) and its zymogen in human placenta

HGF-activator (HGF-A) is a circulating serine protease known to be responsible for activation of hepatocyte growth factor (HGF). Active HGF is thought to be an important regulator of trophoblast growth. In vitro, HGF-A is produced via proteolytic cleavage of its zymogen by thrombin. Immunocytochemistry and Western immunoblotting were performed using human placental tissue from all three trimesters with an antibody that recognizes both HGF-A and its zymogen. Western immunoblotting revealed a 97 kDa band equivalent to the zymogen in placenta from all three trimesters. A smaller 34 kDa band equivalent to HGF-A was only seen in first and second trimester placenta. The anti-HGF-A/zymogen antibody demonstrated immunostaining in placental villi and membranes throughout gestation. Within first trimester villi immunostaining was strongest within the syncytio- and cytotrophoblast layers, but was also seen within stromal and endothelial cells. Likewise, in third trimester placenta the syncytio-cytotrophoblast layer showed the strongest immunoreactivity. In vitro, HGF can induce trophoblast DNA synthesis and the localization of HGF-A to the peri-villous trophoblast layer (which expresses c-met, the HGF receptor) suggests that it may be responsible for activation of pro-HGF at this site. This adds further weight to the hypothesis that HGF in vivo is an important regulator of trophoblast growth.     

The association of arylendosulfatase 1 (SULF1) gene polymorphism with recurrent miscarriage

Purpose

One of the most common problems in reproductive medicine is recurrent miscarriage (RM). There is increasing evidence showing genetic susceptibility of women is an important risk factor in the occurrence of RM. In recent years, there is a growing interest in sulfate and its role in fetal development. A novel mechanism of SULF1 has been demonstrated for modifying the activities of some growth factors and signalling molecules that have major roles during embryogenesis. The aim of present study was to evaluate the association of SULF1 gene polymorphism (rs6990375 G > A) in Iranian patients with RM.

Methods

We established a case-control study of 200 Iranian women: 100 patients with the history of two or more RM as cases and 100 healthy women with at least two cases of successful pregnancy and no history of miscarriage as controls. The polymorphism was examined by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) method.

Results

The genotypic analysis between case and controls showed significant differences (p-value = 0.000). Allelic analysis showed no significant correlation (Χ2 = 3.36, p-value = 0.066). The heterozygous genetic variant was significantly higher among healthy women (OR = 12.67, 95 % CI = 6.47–24.79).

Conclusions

Our data showed that rs6990375 polymorphism of SULF1 gene could be among one of the factors related to RM in Iranian women. Further evaluation of this polymorphism may be important and need further studies.