Overexpression of nerve growth factor in peritoneal fluid from women with endometriosis may promote neurite outgrowth in endometriotic lesions

To investigate the role of the nerve growth factor (NGF) in the endometriosis-associated innervation in the development of endometriosis-associated symptoms, 41 peritoneal fluid samples (PF) from patients with surgically and histologically proven endometriosis and 20 PF from patients with other gynecologic conditions were analyzed with Western blot and a novel in vitro model using dorsal root ganglia (DRG) to show neuronal outgrowth; endometrial cells also were analyzed. The results suggest that the PF of endometriosis patients and endometriotic lesions have neurotropic properties, because the Western blot analysis and the cell culture stainings showed NGF expression, and the neurite outgrowth of DRG treated with PF of patients with endometriosis was significantly higher than when treated with PF of patients without endometriosis. Furthermore, blocking NGF with both anti-NGF and K252a leads to a significant decrease in neurite outgrowth.     

转化生长因子β1与原因不明习惯性流产关系的初步研究

目的:探讨原因不明习惯性流产（UHA）患者主动免疫治疗前后外周血单个核细胞转化生长因子β1（TGF-β1）mRNA水平的变化。方法:用半定量逆转录-聚合酶链反应（RT-PCR）技术,检测正常非孕妇女30例和UHA妇女30例主动免疫治疗前和治疗后3周的外周血单个核细胞内TGF-β1　mRNA 表达水平的相对含量（％）。结果:UHA组外周血单个核细胞TGF-β1　mRNA的相对含量（123.2±62.7）％明显低于正常非孕组妇女（192.7±64.6）％（P<0.01）。主动免疫治疗后,UHA组TGF-β1mRNA的相对含量（172.2±74.9）％较治疗前明显升高（P<0.01）,且和正常非孕组差异无显著性。UHA组经主动免疫治疗后,16例妊娠成功,6例仍发生自然流产。妊娠成功者中TGF-β1　mRNA的相对含量（183.5±90.5）％较治疗前（113.6±68.6）％明显升高（P<0.05）,而妊娠失败者TGF-β1mRNA的相对含量与治疗前无显著差异。结论:亿卜p1表达水平降低可能与UHA的发生相关,主动免疫治疗可能通过上调TGF-β1的表达水平促使UHA患者获得妊娠成功。     

Placental gene expression of transforming growth factor beta 1 (TGF-β1) in small for gestational age newborns

Objective: The gene expression of transforming growth factor beta-1 (TGF-β1) in human placental samples obtained from pregnancies with small for gestational age fetuses (SGA) was compared to those of normal pregnancies.

Methods: In 2011 placental samples from 101 pregnancies with SGA and from 140 normal pregnancies were obtained for analysis of TGF-β1 gene expression. Several clinical parameters were also assessed for correlation between genetic and clinical parameters.

Results: There were no significant differences in gene activity of the TGF-β1 gene between the SGA versus normal pregnancy groups (Ln2^α: 0.16; p?=?0.07). Within the SGA group, no fetal gender-dependent differences were seen in TGF-β1 gene expression (Ln2^α: ?0.11; p?=?0.05). Similarly, no significant differences in gene activity were observed by the degree of severity of SGA as assessed by percentile fetal birth-weight (Ln2^α: 0.32; p?=?0.06).

Conclusion: We found no change in gene expression of TGF-β1 in placental samples obtained from SGA pregnancies versus normal pregnancy suggesting an absence of a direct role of the TGF-β1 gene in the development of SGA. However, the absence of increased gene expression of TGF-β1 in SGA can be conceptualized as a failure to mount a compensatory response in the SGA environment.     

转化生长因子β1在新生未成熟大鼠高氧损伤肺组织中的表达及意义

目的　探讨转化生长因子 β1(TGF β1)在新生未成熟大鼠高氧肺损伤不同时点肺组织中的表达及意义。　方法　将 99只新生未成熟大鼠随机分为高氧组和空气组 ,于日龄 3、7和 14d随机分批处死并取肺组织HE染色进行病理观察 ,免疫组化检测TGF β1在各组中的表达 ,秩和检验组间THF β1阳性强度表达差异。　结果　高氧各组鼠肺组织TGF β1的表达较同期空气组广泛 ,且在肺泡和支气管上皮细胞、肺内间质中的表达强度差别始终强于对照组 (u值分别为 :3d时 :4 9.0、14 .0、6 3.0 ;7d时 :34.0、2 .0、4 5 .0 ;14d时 :32 .0、13.5、33.0 ,P值均 <0 .0 5 ) ,日龄 14d时 ,高氧组鼠肺泡巨噬细胞中可见TGF β1的表达 ,而空气组鼠肺泡巨噬细胞中未见TGF β1的表达 ,两组比较u值为 13.0 ,P <0 .0 5。　结论　肺组织中的TGF β1表达增强以及肺泡巨噬细胞中表达TGF β1,可能与高氧肺损伤有内在联系。     

子痫前期患者胎盘组织中转化生长因子β1、血管细胞黏附分子1及E选择素表达的研究

目的探讨子痫前期患者胎盘组织中转化生长因子β1(TGF-β1)、血管细胞黏附分子1(VCAM-1)和E选择素(E-selectin)的表达变化及其意义。方法用免疫组化链霉菌抗生物素蛋白-过氧化物酶连接(SP)法,对20例正常孕妇(对照组)和40例子痫前期患者(子痫前期组,其中轻度16例、重度24例)的胎盘组织进行TGF-β1、VCAM-1和E-selectin定位,并用计算机图像分析系统进行定量分析比较。结果(1)1FGF-β1在胎盘绒毛合体滋养细胞的表达,子痫前期组为70．7±0．5,对照组为70．3±0．6,两组比较,差异有统计学意义(P<0．05)。VCAM-1的表达,子痫前期组为82．5±0．5,对照组为82．8±0．3;E-selectin的表达,子痫前期组为53．5±0．5,对照组为53．8±0．4;两组分别比较,差异均有统计学意义(P<0．05)。TGF-β1、VCAM-1、E-selectin在轻度子痫前期患者胎盘组织合体滋养细胞中的表达分别为70．6±0．6、82．4±0．6、53．4±0．5,在重度子痫前期患者胎盘组织中的表达分别为70．8±0．4、82．6±0．5、53．6±0．5,轻度与重度子痫前期患者TGF-β1、VCAM-1、E- selectin表达水平比较,差异均无统计学意义(P>0．05)。(2)E-selectin在胎盘绒毛毛细血管内皮细胞的表达,子痫前期组为63．0±0．5,对照组为62．6±0．4,两组比较,差异有统计学意义(P<0．05);轻度子痫前期患者为63．2±0．4、重度子痫前期为62．9±0．5,二者比较,差异无统计学意义(P> 0．05)。结论TGF-β1、VCAM-1和E-selectin在胎盘组织中的表达变化,在子痫前期发病中有重要作用。     

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.     

类固醇生成因子1在卵巢子宫内膜异位囊肿及子宫腺肌病病灶中的表达

目的 探讨类固醇生成因子1( SF-1)在子宫内膜异位症(内异症)患者在位内膜、卵巢内异症囊肿、子宫腺肌病病灶中的表达.方法 选择2008年1月至2010年12月在北京大学第一医院妇科住院,因子宫腺肌病合并卵巢内异症囊肿行子宫全切除术及卵巢内异症囊肿剥除术或附件切除术的患者共30例,经病理确诊子宫腺肌病合并内异症囊肿共17例作为观察组;同期因宫颈上皮内瘤变(CIN)Ⅲ行子宫全切除术的患者10例作为对照组.将观察组患者的在位内膜、卵巢内异症囊肿病灶、子宫腺肌病病灶和对照组的子宫内膜组织进行石蜡切片,免疫组化Envision二步法检测SF-1蛋白的表达.结果 两组患者在位内膜的腺体和间质细胞中均无SF-1蛋白表达;观察组卵巢内异症囊肿病灶的间质细胞核SF-1蛋白的阳性表达率为14/17,SF-1蛋白在卵巢内异症病灶的腺体细胞及子宫腺肌病病灶中均无表达.结论 卵巢内异症囊肿与子宫腺肌病病灶中SF-1蛋白表达的差异可能在疾病的发生与发展中具有重要意义.     

NGF及其受体trkA及p75NTR在子宫内膜异位症患者在位内膜中的表达及其与内异症疼痛的关系

目的:研究子宫内膜异位症(内异症)患者在位内膜神经生长因子(NGF)及其受体trkA、p75NTR的表达,探讨上述因子与内异症疼痛的关系。方法:选择就诊于北京协和医院并进行了腹腔镜手术的28例子宫内膜异位症患者作为研究组,非子宫内膜异位症患者10例作为对照组(B组),收集上述患者分泌期在位子宫内膜。根据患者有无痛经,分为内异症疼痛组(A1组,18例)及内异症非疼痛组(A2组,10例)。采用免疫组化比较各组病灶中NGF及其受体trkA、p75NTR的表达,并分析其与痛经的关系。结果:各组在位内膜腺上皮中NGF的表达显著高于间质(P<0.05);内异症疼痛组腺上皮NGF表达较非疼痛组明显升高(359.9±18.7 vs 201.3±34.3,P<0.05)。p75NTR主要在子宫内膜间质细胞中表达;内异症组明显高于非内异症组(58.8±21.1、22.5±16.1 vs 0,P<0.05);内异症疼痛组较非疼痛组p75NTR表达明显升高(58.8±21.1 vs 22.5±16.1,P<0.05)。trkA在非内异症组子宫内膜间质中表达明显高于腺上皮(P<0.05);在内异症组腺上皮中,trkA的表达较非内异症组明显增高(P<0.05);trkA的表达量与内异症患者疼痛无关。结论:p75NTR可能参与内异症的发病,NGF及其受体p75NTR在在位内膜中的表达与患者疼痛相关,表明其可能参与了内异症疼痛发病机制。     

Comparative evaluation of vascular endothelial growth factor-A expression in pre-ovulatory follicular fluid in normogonadotrophic and endometriotic patients undergoing assisted reproductive techniques

ObjectiveThe aim of this study is to compare the differential expression of VEGF-A in pre-ovulatory follicular fluid of both normogonadotrophic and endometriotic patients undergoing ICSI and its relation to ICSI outcome.SettingAgial IVF – ICSI center, Alexandria, Egypt.MethodologyThe study was a prospective randomized control trial that included 90 infertile patients who were subdivided into two groups:Group A: 45 normogonadotrophic patients as patients having tubal factor and unexplained infertility.Group B: 45 endometriotic patients diagnosed by laparoscopy.All patients underwent the same ovarian hyperstimulation protocol (the long luteal phase protocol). In all patients the blood was sampled on the day of HCG administration. The isolated sera were frozen and stored at −20 °C for later hormone analysis (assay of estradiol and progesterone). The follicular fluid was collected on the day of oocyte retrieval from all the patients. Only aspirates from follicles larger than 17 mm, uncontaminated with blood were included in this study. Follicular fluid samples were centrifuged for 10 min, and the supernatants were stored at −20 °C until hormone measurements were carried out (assay of estradiol, progesterone and androstenedione). VEGF-A assay was performed using the ELISA technique for all the follicular fluid samples and the results were correlated to the ICSI outcome in both groups.Outcome measuresThe primary outcome measures were: (a) The number of mature oocytes per cycle of induction, (b) the grading of embryos obtained, (c) implantation rate, (d) serum level of estradiol and progesterone on the day of HCG administration and (e) follicular fluid level of estradiol, progesterone and androstenedione on the day of oocyte retrieval.The secondary outcome measure was pregnancy rate which was diagnosed by:1. Serum B-hCG assay 14 days after embryo transfer.2. Clinical pregnancy rate.ResultsThere was a statistically significant difference between the two groups when comparing the mean value of the follicular fluid VEGF-A concentrations. In group A, the mean value was 529.4 ± 309.7 (pg/ml), while in group B, it was 1388.7 ± 1152 (pg/ml) (p value = 0.0001). There was no statistical significant difference between both groups as regards the outcome of pregnancy. When correlating between ICSI outcome and different studied parameters a positive correlation was detected between the ICSI outcome and endometrial thickness. No correlation could be detected between the ICSI outcome and other studied parameters. No correlations could be detected between VEGF-A expression and different studied laboratory parameters. However, A positive correlation was detected between VEGF-A and ICSI outcomes in both groups, it means that increasing the level of VEGF-A is accompanied with a decrease in the pregnancy rate (p = 0.003), a negative correlation was detected between VEGF-A and endometrial thickness, a positive correlation was detected between VEGF-A and metaphase I oocytes, otherwise no other correlations could be detected.ConclusionThe current study concluded that follicular fluid VEGF-A is correlated negatively with the ICSI outcome.     

Placental gene expression patterns of epidermal growth factor in intrauterine growth restriction

Objective

In this study, we compared human placental gene expression patterns of epidermal growth factor (EGF) in pregnancies with intrauterine growth restriction (IUGR) vs. normal pregnancies as control.

Study design

Gene expression of EGF was determined from human placental samples collected from all pregnancies presenting with IUGR at our institution during the study period January 1, 2010–January 1, 2011. Multiple clinical variables were also assessed including maternal age, gestational weight gain, increase of BMI during pregnancy and fetal gender.

Results

A total of 241 samples were obtained (101 in the IUGR pregnancy group, 140 in the normal pregnancy group). EGF was found to be underexpressed in the IUGR group compared to normal pregnancy (Ln2^α: −1.54; p < 0.04). Within the IUGR group no fetal gender-dependent difference was seen in EGF gene expression (Ln2^α: 0.44; p < 0.06). Similarly, no significant difference in EGF expression was noted in cases with more vs. less severe forms of IUGR (Ln2^α: −0.08; p = 0.05). IUGR pregnancies were significantly more common in the maternal age group 35–44 years compared to other age groups. Gestational weight gain and gestational BMI increase were significantly lower in IUGR pregnancies compared to controls.

Conclusions

Placental expression of EGF was found to be reduced in IUGR pregnancies vs. normal pregnancies. This may partly explain the smaller placental size and placental dysfunction commonly seen with IUGR. An increased incidence of IUGR was observed with maternal age exceeding 35 years. The probability of IUGR correlated with lower gestational weight gain and lower BMI increase during pregnancy.