雄激素致不孕大鼠胰腺、下丘脑及卵巢雄激素受体mRNA表达

目的:研究雄激素致不孕大鼠(ASR)胰腺、下丘脑及卵巢组织中雄激素受体(AR)mRNA的含量变化。方法:9日龄SD雌性大鼠皮下注射丙酸睾丸酮制备ASR模型,于106日龄左右(动情前期)处死,取血放免法测定△4-雄烯二酮(△4-A)、总睾酮(TT)、游离睾酮(FT)、胰岛素(Ins)和C-肽(C-P),提取胰腺、下丘脑及卵巢总RNA,以单碱基突变模板为内对照的RT-PCR方法对ARmRNA进行定量分析。结果:ASR模型血△4-A、TT、FT、Ins、C-P均显著高于对照组(P<0.05,P<0.01),胰腺、下丘脑及卵巢的AR mRNA表达水平明显升高,与对照组相比差异有统计学意义(P<0.05,P<0.01)。结论:ASR模型血雄激素水平升高,上调胰腺、下丘脑及卵巢AR mRNA的表达,引起该模型的高胰岛素血症和无排卵。     

转化生长因子β1β3及其受体βRⅠβRⅡmRNA在子宫肌瘤组织中的表达

目的　探讨转化生长因子 β1、β3 (TGF β1、β3 )及其受体 βRⅠ、βRⅡmRNA与子宫肌瘤发生发展的关系。方法　 2 0 0 0年 12月至 2 0 0 1年 11月采用免疫组化及原位杂交染色方法 ,对 30例子宫肌瘤及正常子宫肌组织标本检测TGF β1、β3 及其受体 βRⅠ、βRⅡmRNA的表达。 结果　 (1)子宫肌瘤组织中TGF β1、β3 蛋白表达强于邻近正常肌层 (P <0 0 1) ,且黄体期较卵泡期增高 (P <0 0 1)。TGF βRⅠ水平较邻近正常肌层亦增高 (P <0 0 1) ,TGF βRⅡ则下降 (P <0 0 5 )。 (2 )原位杂交TGF β1、β3 mRNA杂交信号的检测结果与免疫组化检测结果一致。结论　TGF β1、β3 与子宫肌瘤的发生发展密切相关 ,TGF βRⅡ降表达或表达异常可能是TGF βs促进子宫肌瘤发生发展的始动环节     

转化生长因子β1及其Ⅱ型受体在卵巢肿瘤中的表达及意义

目的探讨转化生长因子β1(TGF-β1)及其Ⅱ型受体(TGFβR-Ⅱ)在卵巢肿瘤中的表达及其意义。方法1998年7月至2001年12月哈尔滨医科大学第一临床医学院,采用SP免疫组化染色方法,对30例卵巢癌,30例卵巢良性上皮性肿瘤及30例正常卵巢组织中TGF-β1、TGFβR-Ⅱ的表达进行分析。结果卵巢癌组TGF-β1阳性率明显高于正常卵巢组(P<0·01),且二者的强阳性率比较差异有非常显著性意义(P<0·01);卵巢良性上皮性肿瘤中TGF-β1阳性率与正常卵巢组比较,两者的阳性率、强阳性率之间比较差异均无显著性意义(P>0·05)。卵巢癌组TGFβR-Ⅱ阳性率低于正常卵巢组(P<0·05),且两者的强阳性率之间差异有非常显著性意义(P<0·01);卵巢良性上皮性肿瘤中TGFβR-Ⅱ阳性率与正常卵巢组比较,两者的阳性率、强阳性率之间差异均无显著性意义(P>0·05),与卵巢癌组比较,两者的阳性率之间差异有显著性意义(P<0·05),但强阳性率之间差异无显著性意义(P>0·05)。结论在卵巢癌中TGF-β1呈过度表达,使得其抑制癌细胞生长的作用丧失。在卵巢癌中TGFβR-Ⅱ失表达,使肿瘤细胞有效地逃避了机体的抑制。     

转化生长因子β1与多囊卵巢综合征发病的关系

多囊卵巢综合征(PCOS)是妇科最常见的内分泌紊乱性疾病,临床表现多样化,发病率呈逐年上升趋势,但其病因及发病机制目前尚无定论。近年关于PCOS的分子生物学及基因遗传学研究备受关注,国内外学者相继发现转化生长因子β1的异常表达与PCOS的关系密切。已有研究提示,PCOS患者血清及卵泡液中转化生长因子β1的表达高于正常水平,并且在促进卵泡闭锁、卵巢间质纤维化,参与高雄激素血症、高胰岛素血症的形成,以及促进其远期并发症——子宫内膜癌的发生等方面发挥了重要作用。综述PCOS患者体内异常表达的转化生长因子β1对其各个病理改变的影响,以期为临床治疗提供新思路及理论依据。     

碱性成纤维细胞生长因子对新生大鼠缺血脑组织转化生长因子β1及其受体mRNA表达的影响

目的 研究碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)对3日龄新生大鼠脑缺血后星形胶质细胞数目以及转化生长因子-β1(transforming growth factor-β1,TGF-β1)及其受体--Smad2mRNA表达的影响,探讨bFGF对未成熟脑缺血性损伤的保护作用机制. 方法 结扎3日龄新生SD大鼠双侧颈总动脉制备脑缺血模型,随机分为对照组(33只)、治疗组(33只).另取3日龄新生SD大鼠33只为假手术组.免疫荧光染色方法检测三组大鼠术后4 d、7 d和14 d脑室下区胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)阳性细胞数目;实时荧光定量PCR(real-time PCR)技术检测三组大鼠术后4 d、7 d和14 d脑室下区TGF-β1及Srnad2 mRNA表达变化. 结果 (1)假手术组GFAP阳性细胞术后7 d达高峰[(325.4±52.5)个/视野],对照组、治疗组GFAP阳性细胞数术后14 d达高峰[分别为(533.5±75.7)、(727.2±104.5)个/视野)],三组同时点比较差异有统计学意义(P＜0.01).(2)对照组TGF-β1和Smad2 mRNA表达在术后7 d达高峰(分别为7.67±1.22和6.22±1.92),治疗组TGF-β1和Smad2 mRNA表达在术后14 d达高峰(分别为8.65±1.02和7.67±1.41),三组同时点比较差异有统计学意义(P＜0.01). 结论 未成熟脑缺血后,外源性bFGF可通过诱导TGF-β1的表达,引起星形胶质细胞反应性增生,而发挥其神经营养作用.     

转化生长因子β及其受体在胚胎卵巢中的分布和作用

转化生长因子β(transforming growth factor-β，TGF-β)属于低分子多肽生长因子，可分为5种(TGF-β1～TGF-β5)，是一种具有多种生物学功能的细胞因子，其不仅在成熟组织和肿瘤组织中有表达，胚胎发育过程中各种胚胎组织细胞中也存在着TGF-β和TGF-βR的表达，它不仅广泛参与哺乳类动物的各种病理生理活动，在胚胎发育中也起着重要作用。一般来说，其对问充质来源的细胞起增殖作用，而对上皮或神经外胚层来源的细胞起抑制作用。本文将主要就TGF-β及其受体在不同发育阶段的胚胎卵巢中的表达、功能综述如下。     

雌孕激素受体及转化生长因子β1在宫腔粘连发病机制中的作用

目的通过对子宫内膜组织中雌激素受体(ER)、孕激素受体(PR)及转化生长因子β1(TGF-β1)表达的研究,探讨宫腔粘连的发病机制。方法2002年7月至2003年12月首都医科大学附属复兴医院以前瞻性方法观察宫腔粘连组(观察组)和非宫腔粘连组(对照组)患者各30例,采用酶联免疫测定法,测定两组血清6项性激素:包括卵泡刺激素(FSH)、黄体生成素(LH)、雌激素(E2)、催乳素(PRL)、孕酮(P)及睾酮(T)。通过免疫组化半定量测定两组子宫内膜组织中ER、PR及TGF-β1的表达水平。结果两组血清性激素水平FSH、LH、E2、PRL、P及T差异无显著性意义(P>0·05);观察组子宫内膜TGF-β1和ER的表达明显高于对照组(P值分别为<0·01,<0·05);在观察组中重度宫腔粘连患者子宫内膜TGF-β1的表达明显高于轻、中度宫腔粘连患者,两者间子宫内膜TGF-β1表达差异有非常显著性意义(P<0·01);两组子宫内膜腺上皮细胞PR的表达差异无显著性意义(P>0·05)。结论子宫内膜组织中ER和TGF-β1的异常表达可能参与宫腔粘连的形成。     

转化生长因子β及其受体与卵泡发育

转化生长因子β(TGF-β)是一类具有多种生物学活性的细胞因子,而卵巢中的卵泡发育是一动态和复杂的过程.对不同标本进行研究证明,TGF-β及其受体在卵巢不同发育阶段中均可表达,通过自分泌/旁分泌机制参与调控卵巢功能,对维持卵巢内环境稳定发挥重要作用.     

转化生长因子β1及其受体与子宫内膜癌发生及临床病理参数关系的探讨

目的 :探讨转化生长因子β1(transforminggrowthfactorβ1,TGFβ1)及其Ⅰ、Ⅱ型受体TGFβR Ⅰ、TGFβR Ⅱ与子宫内膜癌发生及临床病理参数的关系。 方法 :采用免疫组化多克隆抗体技术检测 30份正常子宫内膜癌组织、15份子宫内膜复合增生组织 (其中 9份有不典型增生 )、12份正常子宫内膜组织中TGFβ1及其受体TGFβR Ⅰ、TGFβR Ⅱ的表达。结果 :子宫内膜癌及复合增生组织中TGFβ1表达明显高于正常子宫内膜组织中者 (P<0 .0 1,P <0 .0 5) ,而前两者之间差异无显著性 (P >0 .0 5) ,且TGFβ1表达水平与子宫内膜癌肌层浸润深度呈正相关 (P <0 .0 5)。从正常子宫内膜、复合增生到子宫内膜癌组织中TGFβR Ⅰ、TGFβR Ⅱ表达逐渐下降甚至缺如 ,且两两之间差异均有显著性 (P <0 .0 5)。TGFβR Ⅰ、TGFβR Ⅱ均与肌层浸润深度呈负相关 (P <0 .0 5)。 结论 :TGFβ1过度表达可能促进子宫内膜癌的进展 ,可能是子宫内膜癌发生的早期现象 ,其负性调控的丧失与TGFβR Ⅰ、TGFβR Ⅱ表达下降或缺如有关。     

转化生长因子β及其受体与卵泡发育

转化生因子β（TGF－β）是一类具有多种生物学活性的细胞因子,而卵巢中的卵泡发育是一动态和复杂的过程。对不同标本进行研究证明,TGF－β及其受体在卵巢不同发育阶段中均可表达,通过自分泌／旁分泌机制参与调控卵巢功能,对维持卵巢内环境稳定发挥重要作用。     

雌、孕激素受体、生长激素受体、胰岛素样生长因子1受体与子宫内膜容受性的关系

辅助生殖技术(ART)经过近40年的发展,已取得突飞猛进的进展。但是临床妊娠率仍然徘徊在30%~40%。决定能否成功妊娠的因素主要包括2个方面,一是胚胎发育的质量,二是子宫内膜对胚胎的容受性。而在临床工作中,即使胚胎质量良好,种植率仍然不能令人满意。因此,如何提高子宫内膜容受性已经成为辅助生殖领域一个亟待解决的问题,了解影响子宫内膜容受性的相关因素,对于在种植窗期如何调节母体的内膜达到最佳的接纳胚胎的状态,具有重要的指导性意义。本文着重阐述子宫内膜容受性与雌激素受体(ER)、孕激素受体(PR)、胰岛素样生长因子1受体(IGF-1R)、生长激素受体(GHR)之间的关系,期望对临床判断子宫内膜容受性有所帮助。     

A variant of the glucocorticoid receptor gene is not associated with adrenal androgen excess in women with polycystic ovary syndrome

Objective: To determine whether the frequency of the N363S variant of the glucocorticoid receptor (GRL) was increased in women with PCOS and adrenal androgen (AA) excess.

Design: Prospective case-control study.

Setting: University reproductive endocrinology laboratory and outpatient clinic.

Patient(s): Consecutive patients of non-Hispanic white race diagnosed with PCOS (n = 114) and healthy controls (n = 92).

Intervention(s): Blood and DNA sampling before hormonal therapy.

Main Outcome Measure(s): PCOS patient and healthy control genotypes, with the N363S allele representing a variant of GRL.

Result(s): Fifty-four PCOS patients with (DHEAS ≥ 3000 ng/mL) and 55 without (DHEAS ≤ 2,500 ng/mL) AA excess, respectively, were studied. Six of 109 (5.5%) patients studied were found to be heterozygous carriers of the A→G base pair substitution at cDNA position 1220, resulting in the missense mutation N363S. Of these six, four had excessive AA secretion (i.e., excess DHEAS levels). There was no significant difference in the allele frequency of the GRL variant between PCOS patients with and without AA excess and controls (3.7% [95% confidence interval: 1.0%–5.7%], 1.8% [0.2%–6.0%], and 3.3% [2.3%–6.0%]). None of the subjects were found to be homozygous for the N363S allele.

Conclusion(s): The N363S variant of GRL was an uncommon occurrence in our population of healthy women and PCOS patients and did not appear to play a major role in the genetic predisposition to PCOS or to AA excess in PCOS.     

TGF-β_1在PCOS卵巢间质纤维化形成中的作用

目的:探讨TGF-β1在PCOS大鼠卵巢间质纤维化、包膜硬化形成中的作用。方法:利用脱氢表雄酮(DHEA)皮下注射的方法建立PCOS病理模型大鼠20只,用微粒子酶免分析法测定血清性激素E2、T、LH、FSH、LH/FSH、空腹胰岛素(FINS)水平,及光镜下观察PCOS大鼠卵巢的病理结构,透射电镜观察细胞超微结构来验证模型。采用免疫组化法检测PCOS组(n=20)卵巢细胞因子TGF-β1的表达,并与20只正常大鼠相比照。结果:TGF-β1在PCOS组各阶段卵泡卵母细胞、颗粒细胞的表达强度与对照组相比,差异无统计学意义(P>0.05)。而在窦状卵泡膜细胞和卵巢间质细胞中的表达,PCOS组显著高于对照组(P<0.01,P<0.05)。结论:多囊卵巢中TGF-β1表达异常,可能是导致多囊卵巢间质纤维化、包膜增厚的原因之一,TGF-β1也参与PCOS卵泡发育、闭锁的调控。     

Prevalence of CYP21 mutations and IRS1 variant among women with polycystic ovary syndrome and adrenal androgen excess

OBJECTIVE: To determine whether frequencies of the mutations in the 21-hydroxylase (CYP21) gene and the G972R variant of the insulin receptor substrate-1 (IRS1) gene are increased in women with polycystic ovary syndrome (PCOS) and adrenal androgen (AA) excess. DESIGN: Prospective case-control study. SETTING: University reproductive endocrinology laboratory and outpatient clinic. PATIENT(S): Consecutive patients of non-Hispanic white race diagnosed with PCOS (n = 114) and healthy controls (n = 95). INTERVENTION(S): Blood and DNA sampling before hormonal therapy. MAIN OUTCOME MEASURE(S): Polycystic ovary syndrome patient and healthy control genotypes, with the CYP21 and IRS1 variants. RESULT(S): Fifty-four PCOS patients with (DHEAS >3000 ng/mL) and 55 without (DHEAS <2500 ng/mL) AA excess, respectively, were studied. Of 109 patients studied, 16 (14.7%) were found to be heterozygous carriers of mutations in the CYP21 gene. Of these 16, 10 (62.5%) had excessive AA secretion (i.e., excess DHEAS levels). Fifteen patients (13.8%) were found to be heterozygous carriers of the IRS1 variant; 9 (60.0%) of these 15 had excessive AA secretion. There were no significant differences in the allele frequency of CYP21 mutations or the IRS1 variant between PCOS patients with and without AA excess, and controls. None of the subjects were found to be homozygous carriers of CYP21 mutations or the IRS1 variant. Combined heterozygosity for CYP21 mutations and the IRS1 variant was limited to women with PCOS and excessive AA (n = 3). CONCLUSION(S): The G972R variant of the IRS1 gene might represent a modifier locus among women who are heterozygous carriers of CYP21 mutations, potentially increasing their risk of developing AA excess in PCOS. Nonetheless, this IRS1 variant and CYP21 mutations seem to play a limited role in the development of PCOS in the population studied.     

Vascular endothelial growth factor and its soluble receptor in patients with polycystic ovary syndrome undergoing IVF

We aimed to examine the behaviour of the angiogenetic factor vascular endothelial growth factor (VEGF) and its soluble receptor (sVEGFR-1) in polycystic ovary patients undergoing In vitro fertilisation (IVF) compared with respect to normally ovulating controls. Levels of VEGF and sVEGFR-1 were compared in follicular fluid and serum, both on the day of human choriogonadotropin (hCG) administration and on the day of oocyte retrieval (OR), in controls and polycystic ovarian syndrome (PCOS) patients undergoing IVF cycles. The bioactivity of VEGF (VEGF/sVEGFR-1 ratio) in the two groups was calculated.

Thirty PCOS patients and 20 controls referring to the IVF Centre of the University of Pisa (Italy) were enrolled. In each patient, blood samples were collected on the day of hCG and on the day of OR administration, and follicular fluid samples. VEGF and sVEGFR-1 were measured by Enzyme Linked Immuno Sorbant Assay (ELISA).

Serum VEGF bioactivity markedly increased in both groups after hCG administration. Serum and follicular fluid VEGF bioactivity was greater in PCOS patients than in controls on the day of OR. The increase in VEGF bioactivity in PCOS patients undergoing IVF was not only because of increasing levels of VEGF but also to decreasing levels of its soluble receptor. We believe that additional studies will clarify their role in the pathogenesis of ovarian hyperstimulation syndrome, which most often occurs in patients with PCOS.     

Raised serum levels of matrix metalloproteinase-9 in women with polycystic ovary syndrome and its association with insulin-like growth factor binding protein-1

Background.?It has been suggested in recent studies that matrix metalloproteinases (MMPs) may be implicated in the pathogenesis of polycystic ovary syndrome (PCOS) through regulating ovarian tissue remodeling. In addition to degrading the extracellular matrix, MMPs exhibit the ability to cleave insulin-like growth factor binding protein-1 (IGFBP-1), the major regulator of insulin-like growth factor-I (IGF-I) in serum. The present study aimed to investigate the possible role of MMPs in the pathophysiology of PCOS.

Methods.?Serum levels of MMP-9, tissue inhibitor of metalloproteinase-1 (TIMP-1), IGF-I and IGFBP-1 were measured in 42 patients with PCOS and 30 healthy women with regular menstruation, matched for age and body mass index. Correlation between IGFBP-1 and other parameters in the PCOS group was analyzed by Pearson's linear correlations.

Results.?Serum MMP-9 concentrations and MMP-9/TIMP-1 ratios were significantly higher in PCOS women than in controls. Serum levels of IGFBP-1 were markedly lower in the PCOS group. There was a negative correlation between serum IGFBP-1 and MMP-9 in women with PCOS.

Conclusion.?Our results raise the possibility that MMPs may be implicated in the pathophysiology of PCOS either by regulating ovarian tissue remodeling or indirectly by facilitating IGF-I bioavailability through proteolysis of IGFBP-1.     

白细胞介素6对雄激素活性的影响及在多囊卵巢综合征发病机制中的作用

目的探讨白细胞介素6(IL-6)对雄激素活性的影响及在多囊卵巢综合征(PCOS)发病机制中的作用。方法在体外培养的猪卵泡内膜细胞和颗粒细胞培养液中分别加入IL-6(10ng/L、100ng/L、1000ng/L),观察IL-6对卵泡内膜细胞睾酮分泌及卵巢颗粒细胞雄激素受体(AR)mRNA表达的影响。结果在24h、48h、72h三个时间点分别与空白对照组(0ng/L)相比,加入IL-6后卵泡内膜细胞睾酮分泌无明显变化;在72h时通过Real-timePCR检测发现,IL-6(100ng/L、1000ng/L)对卵巢颗粒细胞AR-mRNA的表达具有上调作用。结论IL-6虽然没有直接促进卵巢睾酮分泌的作用,但却可以上调卵巢组织AR-mRNA的表达,间接增强雄激素活性,此可能与慢性亚临床炎症可引发PCOS的机制有关。     

The serum level of transforming growth factor beta1 and its association with Foxp3 gene polymorphism in Iranian women with recurrent spontaneous abortion

The aim of the study was to determine the role of transforming growth factor-beta1 (TGF-β1) and Foxp3 (rs3761548) promoter polymorphisms in Iranian women with recurrent spontaneous abortion (RSA). Eighty women with RSA were compared with eighty in a control group. Serum levels of TGF-β1 were measured using ELISA and Foxp3 (rs3761548) promoter polymorphisms using a PCR-RFLP technique. In addition, serum levels of TGF-β1 were compared in different genotypes in the two groups. The women's ages in the two groups were similar (30.15 ± 4.42 years [RSA] vs. 29.97 ± 4.51 [control]) as were serum TGF-β1 concentrations in case and control groups (53.42 ± 2.08 ng/ml in control and 56.31 ± 2.58 ng/ml in the RSA group; p = 0.4). Furthermore, there was no significant difference in the genotype frequencies of the rs3761548 Foxp3 gene between the two groups (p = 0.3) and the levels of TGF-β1 were similar in different genotypes. In conclusion, the data indicate that serum TGF-β1 levels and Foxp3 (rs3761548) promoter polymorphism is not a risk factor for RSA and that there is no association between the polymorphism and serum TGF-β1 levels.