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Aquaporin 9 regulates Leydig cell steroidogenesis in diabetes 水通道蛋白9调节糖尿病患者睾丸间质细胞类固醇生成
IF 2.4 4区 医学 Q3 ANDROLOGY Pub Date : 2022-03-30 DOI: 10.1080/19396368.2022.2033350
Arun K. Kannan, Lezy Flora Mariajoseph-Antony, Antojenifer Panneerselvam, Chithra Loganathan, Diwakar Kiduva Jothiraman, K. Anbarasu, C. Prahalathan
Abstract Diabetes mellitus induced hyperglycemia increases oxidative stress, which contributes to impairment of male reproductive function. Aquaporins (AQPs) belong to a transmembrane protein superfamily containing 13 isoforms (AQP0-12), differentially expressed in various organs, and play a pivotal role in male reproductive function. In the current study, we investigated the relationship between AQPs and testicular steroidogenesis under hyperglycemia in vivo and in vitro. The effect of high glucose on the role of AQPs in Leydig cell steroidogenesis was analyzed in diabetic rats (in-vivo) and LC540 rat Leydig cells (in vitro) via enzyme assays, quantitative RT-PCR, siRNA knock down and western blotting. AQP 9 was significantly up-regulated in STZ-induced diabetic rat testis and high glucose treated LC540 cells. Further, oxidative stress marker nuclear factor erythroid 2-related factor 2 (Nrf2) expression was decreased with impaired testicular steroidogenesis under hyperglycemia. Knock-down of AQP 9 resulted in increased Nrf2 expression and thus increased testicular steroidogenesis in hyperglycemia. Diabetes-associated hyperglycemia induced oxidative stress is a widely proven cause for diabetes-related male infertility. Our results collectively suggest that AQP 9 impairs testicular steroidogenesis via the regulation of oxidative stress in diabetes.
摘要糖尿病引起的高血糖会增加氧化应激,从而导致男性生殖功能受损。水通道蛋白(AQPs)属于一个跨膜蛋白超家族,包含13种异构体(AQP0-12),在不同器官中差异表达,在男性生殖功能中发挥着关键作用。在本研究中,我们在体内外研究了高血糖条件下AQPs与睾丸类固醇生成的关系。通过酶分析、定量RT-PCR、siRNA敲除和蛋白质印迹,分析了高糖对AQPs在糖尿病大鼠(体内)和LC540大鼠Leydig细胞(体外)中的作用的影响。AQP9在STZ诱导的糖尿病大鼠睾丸和高糖处理的LC540细胞中显著上调。此外,氧化应激标志物核因子-红系2相关因子2(Nrf2)的表达随着高血糖下睾丸类固醇生成受损而降低。AQP 9的敲除导致Nrf2表达增加,从而在高血糖中增加睾丸类固醇生成。糖尿病相关的高血糖诱导的氧化应激是糖尿病相关男性不育的广泛原因。我们的研究结果共同表明,AQP 9通过调节糖尿病患者的氧化应激来损害睾丸类固醇生成。
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引用次数: 4
Racial/ethnic disparities in infertility treatment utilization in the US, 2011–2019 2011-2019年美国不孕不育治疗利用率的种族/民族差异
IF 2.4 4区 医学 Q3 ANDROLOGY Pub Date : 2022-03-28 DOI: 10.1080/19396368.2022.2038718
Deepa Dongarwar, Vicki Mercado-Evans, Sylvia Adu-Gyamfi, Mei-Li Laracuente, H. Salihu
Abstract With delayed child-bearing age, there has been an increase in infertility rates globally and in the United States (US). Unsurprisingly, there has been a concomitant substantial increase in the number of individuals seeking infertility treatments over the last decade. This study aimed to examine the relationship between race/ethnicity and the utilization of different infertility treatments over the previous decade. We conducted this retrospective cohort study using the United States (US) Birth data files 2011–2019. We calculated the rates of infertility treatment and its subtypes over the study period. Descriptive statistics were utilized to examine the sociodemographic and birth characteristics for overall births and those associated with any infertility treatment and each of its subtypes. We calculated the level of association between race/ethnicity and utilization of infertility treatment and the subtypes using adjusted logistic regression models. We found that the rate of infertility treatments for all subtypes considered, had steadily increased by 63.7% within the past decade. In contrast, fertility enhancing drugs or Intrauterine Insemination (IUI) increased by 134%, and in vitro fertilization (IVF), gamete intrafallopian transfer (GIFT), and zygote intrafallopian transfer (ZIFT) treatments increased by 40% over the 9-year study period. Non-Hispanic (NH) Asian women had the highest rate of any infertility treatment with a rate of 25 per 1000 births whereas Hispanic women had the lowest rate of any infertility treatment at 5.8 per 1000 births. When compared with NH-White women, NH-Asian women had a modest 7% lower likelihood (OR = 0.93, 95% CI = 0.92–0.94) of receiving any infertility treatment while NH-Black and Hispanic women had about 70% lower likelihood of receiving any infertility treatment. Our report of increased assisted reproductive technology (ART) utilization rates, and marked racial/ethnic differences in ART utilization highlight the importance of expanding knowledge of inequities that continue to impact marginalized groups, a critical step for informing actionable strategy formulations (i.e., advocacy, policy change, patient education, provider training) to address these inequities.
摘要随着生育年龄的推迟,全球和美国的不孕不育率都在上升。不出所料,在过去十年中,寻求不孕不育治疗的人数也随之大幅增加。这项研究旨在检验种族/民族与过去十年中不同不孕不育治疗方法的使用之间的关系。我们使用美国2011-2019年出生数据文件进行了这项回顾性队列研究。我们计算了研究期间不孕不育的治疗率及其亚型。描述性统计用于检查总体出生的社会人口统计学和出生特征,以及与任何不孕不育治疗及其每种亚型相关的社会人口统计和出生特征。我们使用调整后的逻辑回归模型计算了种族/民族与不孕不育治疗利用率和亚型之间的关联水平。我们发现,在过去十年中,所有亚型的不孕不育治疗率稳步上升了63.7%。相比之下,在9年的研究期间,提高生育能力的药物或宫内节育器(IUI)增加了134%,体外受精(IVF)、配子输卵管内移植(GIFT)和受精卵输卵管内转移(ZIFT)治疗增加了40%。非西班牙裔(NH)亚洲妇女的不孕不育治疗率最高,为每1000名新生儿中有25名接受不孕不育治疗,而西班牙籍妇女的不孕治疗率最低,为每千名新生儿中5.8名。与NH白人女性相比,NH亚裔女性接受任何不孕治疗的可能性略低7%(OR=0.93,95%CI=0.92–0.94),而NH黑人和西班牙裔女性接受任何不育治疗的可能性约低70%。我们关于辅助生殖技术利用率提高以及辅助生殖技术使用方面明显的种族/民族差异的报告强调了扩大对继续影响边缘化群体的不平等现象的认识的重要性,为解决这些不公平现象提供可操作的战略制定(即宣传、政策变化、患者教育、提供者培训)的关键步骤。
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引用次数: 5
Regulation of STUB1 expression and its biological significance in mouse Sertoli cells 小鼠支持细胞中STUB1的表达调控及其生物学意义
IF 2.4 4区 医学 Q3 ANDROLOGY Pub Date : 2022-03-26 DOI: 10.1080/19396368.2022.2027554
Tao Li, Chao Zheng, W. Han, Zhen-Zhen Chen
Abstract STIP1 Homology and U-Box Containing Protein 1 (STUB1), a ubiquitin E3 ligase initially involved in immune responses, has recently emerged as a pleiotropic regulator of different biological systems, including skeletal and male reproduction systems. On the latter, a homozygous mutation in the STUB1 gene has been identified in patients with hypogonadism. However, the pattern of expression and biological actions of STUB1 in testis remains so far unexplored. Herein, we report analyses on the testicular expression of STUB1 in human testes with impaired spermatogenesis and paracrine regulation of STUB1 expression in mouse testis development and the direct effects of ablation STUB1 on Sertoli cell (SC) functions. STUB1 was expressed abundantly in pachytene spermatocytes and SCs, and weakly in spermatogonia and differentiating spermatids in normal human testis. In contrast, Sertoli-specific expression of STUB1 was significantly decreased in the human testes with impaired spermatogenesis. Throughout postnatal development of mouse testis, however, STUB1 was expressed exclusively in the nuclei of the functionally mature SCs. The adjacent germ cell (GC)-derived IL-1α overtly regulated STUB1 expression through promoting the ETS domain transcription factor Elk-1 (ELK1)-mediated transactivation. Importantly, ablation of endogenous STUB1 caused lipid accumulation and senescence in GC co-incubated SCs. Together with previous reports on the stimulatory effects of IL-1α on cell senescence, our findings suggest that STUB1 may serve as an important negative feedback signaling to modulate the magnitude of GCs-derived IL-1α, which is normally maintained at low levels within testis.
STIP1同源性和U-Box Containing Protein 1 (STUB1)是一种泛素E3连接酶,最初参与免疫应答,最近被发现是不同生物系统的多效调节因子,包括骨骼和男性生殖系统。关于后者,在性腺功能减退症患者中发现了STUB1基因的纯合突变。然而,迄今为止,STUB1在睾丸中的表达模式和生物学作用仍未被探索。在此,我们报道分析了精子发生受损的人睾丸中STUB1的表达、小鼠睾丸发育中STUB1表达的旁分泌调节以及消融STUB1对支持细胞(SC)功能的直接影响。STUB1在正常人睾丸粗线精母细胞和sc中大量表达,而在精原细胞和分化精母细胞中表达较弱。相比之下,在精子发生受损的人类睾丸中,sertoli特异性表达STUB1显著降低。然而,在小鼠睾丸出生后的整个发育过程中,STUB1仅在功能成熟的sc的细胞核中表达。邻近生殖细胞(GC)来源的IL-1α通过促进ETS结构域转录因子Elk-1 (ELK1)介导的反激活来调节STUB1的表达。重要的是,内源性STUB1的消融导致GC共孵育SCs的脂质积累和衰老。结合之前关于IL-1α对细胞衰老的刺激作用的报道,我们的研究结果表明,STUB1可能作为一个重要的负反馈信号来调节gcs来源的IL-1α的大小,而在睾丸内,IL-1α通常维持在低水平。
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引用次数: 2
Is there a role for small molecule metabolite biomarkers in the development of a diagnostic test for endometriosis? 小分子代谢物生物标志物在子宫内膜异位症诊断测试的发展中是否有作用?
IF 2.4 4区 医学 Q3 ANDROLOGY Pub Date : 2022-03-04 DOI: 10.1080/19396368.2022.2027045
Nicola E Tomkins, J. Girling, B. Boughton, S. Holdsworth-Carson
Abstract Endometriosis is a disease defined by the presence of benign lesions of endometrial-like glands and stroma outside the endometrial cavity. Affecting an estimated 11.4% of Australian women, symptoms include chronic pelvic pain, dysmenorrhea and infertility. The current gold standard of diagnosis requires an expensive and invasive laparoscopic surgery, resulting in delayed time to treatment. The identification of a non-invasive endometriosis biomarker – a measurable factor correlating with disease presence or activity – has therefore become a priority in endometriosis research, although no biomarker has yet been validated. As small molecule metabolites and lipids have emerged as a potential focus, this review with systematic approach, aims to summarize studies examining metabolomic biomarkers of endometriosis in order to guide future research. EMBASE, PubMed and Web of Science were searched using keywords: lipidomics OR metabolomics OR metabolome AND diagnostic tests OR biomarkers AND endometriosis, and only studies written in English from August 2000 to August 2020 were included. Twenty-nine studies met inclusion and exclusion criteria and were included. These studies identified potential biomarkers in serum, ectopic tissue, eutopic endometrium, peritoneal fluid, follicular fluid, urine, cervical swabs and endometrial fluid. Glycerophospholipids were identified as potential biomarkers in all specimens, except urine and cervical swab specimens. However, no individual molecule or metabolite combination has reached clinical diagnostic utility. Further research using large study populations with robust patient phenotype and specimen characterisation is required if we are to make progress in identifying and validating a non-invasive diagnostic test for endometriosis.
子宫内膜异位症是一种由子宫内膜腔外存在子宫内膜样腺体和间质的良性病变所定义的疾病。估计有11.4%的澳大利亚妇女受到影响,其症状包括慢性盆腔疼痛、痛经和不孕症。目前的黄金诊断标准需要昂贵的侵入性腹腔镜手术,导致治疗时间延迟。因此,确定一种非侵入性子宫内膜异位症生物标志物——一种与疾病存在或活动相关的可测量因素——已成为子宫内膜异位症研究的重点,尽管尚未有生物标志物得到验证。由于小分子代谢物和脂质已成为潜在的研究重点,本文旨在系统总结子宫内膜异位症代谢组学生物标志物的研究,以指导未来的研究。使用关键词:脂质组学或代谢组学或代谢组学、诊断测试或生物标志物和子宫内膜异位症对EMBASE、PubMed和Web of Science进行检索,仅包括2000年8月至2020年8月期间用英文撰写的研究。29项研究符合纳入和排除标准并被纳入。这些研究在血清、异位组织、异位子宫内膜、腹膜液、卵泡液、尿液、宫颈拭子和子宫内膜液中发现了潜在的生物标志物。除尿液和宫颈拭子标本外,甘油磷脂在所有标本中被鉴定为潜在的生物标志物。然而,没有单独的分子或代谢物组合达到临床诊断的效用。如果我们要在确定和验证子宫内膜异位症的非侵入性诊断测试方面取得进展,就需要使用具有强大患者表型和标本特征的大型研究人群进行进一步的研究。
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引用次数: 1
Cumulus cell acetyl-CoA metabolism from acetate is associated with maternal age but only partially with oocyte maturity. 卵丘细胞乙酰辅酶a代谢与母体年龄有关,但与卵母细胞成熟度仅部分相关。
IF 2.4 4区 医学 Q3 ANDROLOGY Pub Date : 2022-02-01 DOI: 10.1080/19396368.2021.2003479
Sharon Anderson, Peining Xu, Alexander J Frey, Jason R Goodspeed, Mary T Doan, John J Orris, Nicolle Clements, Michael J Glassner, Nathaniel W Snyder

Cumulus cell (CC) clumps that associate with oocytes provide the oocytes with growth and signaling factors. Thus, the metabolism of the CCs may influence oocyte function, and CC metabolism may be predictive of oocyte competence for in vitro fertilization. CCs are thought to be highly glycolytic, but data on the use of other potential carbon substrates are lacking in humans. This prospective and blinded cohort study was designed to examine the substrate utilization of CCs by age and oocyte competence. Individual sets of CC clumps from participants were removed after oocyte retrieval procedure then, incubated with stable isotope labeled substrates, and analyzed using liquid chromatography-high resolution mass spectrometry (LC-HRMS) for isotopologue enrichment of major metabolic intermediates, including acetyl-CoA. The acyl-chain of acetyl-CoA contains 2 carbons that can be derived from 13C-labeled substrates resulting in an M + 2 isotopologue that contains 2 13C atoms. Comparing the fate of three major carbon sources, mean enrichment of M + 2 acetyl-CoA (mean, standard deviation) was for glucose (3.6, 7.7), for glutamine (9.4, 6.2), and for acetate (20.7, 13.9). Due to this unexpected high and variable labeling from acetate, we then examined acetyl-CoA mean % enrichment from acetate in 278 CCs from 21 women ≤34 (49.06, 12.73) decreased with age compared to 124 CCs from 10 women >34 (43.48, 16.20) (p = 0.0004, t-test). The CCs associated with the immature prophase I oocytes had significantly lower enrichment in M + 2 acetyl CoA compared to the CCs associated with the metaphase I and metaphase II oocytes (difference: -6.02, CI: -1.74,-13.79, p = 0.013). Acetate metabolism in individual CC clumps was positively correlated with oocyte maturity and decreased with maternal age. These findings indicate that CC metabolism of non-glucose substrates should be investigated relative to oocyte function and age-related fertility.Abbreviations: CCs: cumulus cells; COC: cumulus-oocyte complex; LC-MS: liquid chromatography-mass spectrometry; acetyl-CoA: acetyl-Coenzyme A; CoA: Coenzyme A.

卵丘细胞(CC)团块与卵母细胞相关,为卵母细胞提供生长和信号因子。因此,CC的代谢可能影响卵母细胞的功能,而CC的代谢可能预测卵母细胞体外受精的能力。CCs被认为具有高度的糖酵解作用,但缺乏关于人类使用其他潜在碳底物的数据。这项前瞻性和盲法队列研究旨在通过年龄和卵母细胞能力来检查cc的底物利用率。在卵母细胞回收程序后,从参与者身上分离出单独的CC团块,与稳定同位素标记的底物孵育,并使用液相色谱-高分辨率质谱(LC-HRMS)分析主要代谢中间体(包括乙酰辅酶a)的同位素富集。乙酰辅酶a的酰基链含有2个碳,可以从13C标记的底物中得到,从而形成含有2个13C原子的M + 2同位素。比较三种主要碳源的命运,M + 2乙酰辅酶a的平均富集(平均值,标准差)为葡萄糖(3.6,7.7),谷氨酰胺(9.4,6.2)和乙酸(20.7,13.9)。由于这一意想不到的高和可变标记,我们随后检查了来自21名≤34岁女性(49.06,12.73)的278个cc中乙酰辅酶a的平均百分比随着年龄的增长而下降,而来自10名>34岁女性(43.48,16.20)的124个cc中乙酰辅酶a的平均百分比随着年龄的增长而下降(p = 0.0004, t检验)。与中期I和中期II卵母细胞相关的cc相比,未成熟前期I卵母细胞相关的cc中M + 2乙酰辅酶a的富集显著降低(差异:-6.02,CI: -1.74,-13.79, p = 0.013)。单个CC团块的醋酸盐代谢与卵母细胞成熟度呈正相关,随母亲年龄的增加而降低。这些发现表明,非葡萄糖底物的CC代谢应该与卵母细胞功能和年龄相关的生育能力进行研究。缩写词:CCs:积云细胞;COC:卵母细胞复合体;LC-MS:液相色谱-质谱法;乙酰辅酶A:乙酰辅酶A;CoA:辅酶A。
{"title":"Cumulus cell acetyl-CoA metabolism from acetate is associated with maternal age but only partially with oocyte maturity.","authors":"Sharon Anderson,&nbsp;Peining Xu,&nbsp;Alexander J Frey,&nbsp;Jason R Goodspeed,&nbsp;Mary T Doan,&nbsp;John J Orris,&nbsp;Nicolle Clements,&nbsp;Michael J Glassner,&nbsp;Nathaniel W Snyder","doi":"10.1080/19396368.2021.2003479","DOIUrl":"https://doi.org/10.1080/19396368.2021.2003479","url":null,"abstract":"<p><p>Cumulus cell (CC) clumps that associate with oocytes provide the oocytes with growth and signaling factors. Thus, the metabolism of the CCs may influence oocyte function, and CC metabolism may be predictive of oocyte competence for in vitro fertilization. CCs are thought to be highly glycolytic, but data on the use of other potential carbon substrates are lacking in humans. This prospective and blinded cohort study was designed to examine the substrate utilization of CCs by age and oocyte competence. Individual sets of CC clumps from participants were removed after oocyte retrieval procedure then, incubated with stable isotope labeled substrates, and analyzed using liquid chromatography-high resolution mass spectrometry (LC-HRMS) for isotopologue enrichment of major metabolic intermediates, including acetyl-CoA. The acyl-chain of acetyl-CoA contains 2 carbons that can be derived from <sup>13</sup>C-labeled substrates resulting in an M + 2 isotopologue that contains 2 <sup>13</sup>C atoms. Comparing the fate of three major carbon sources, mean enrichment of M + 2 acetyl-CoA (mean, standard deviation) was for glucose (3.6, 7.7), for glutamine (9.4, 6.2), and for acetate (20.7, 13.9). Due to this unexpected high and variable labeling from acetate, we then examined acetyl-CoA mean % enrichment from acetate in 278 CCs from 21 women ≤34 (49.06, 12.73) decreased with age compared to 124 CCs from 10 women >34 (43.48, 16.20) (p = 0.0004, t-test). The CCs associated with the immature prophase I oocytes had significantly lower enrichment in M + 2 acetyl CoA compared to the CCs associated with the metaphase I and metaphase II oocytes (difference: -6.02, CI: -1.74,-13.79, p = 0.013). Acetate metabolism in individual CC clumps was positively correlated with oocyte maturity and decreased with maternal age. These findings indicate that CC metabolism of non-glucose substrates should be investigated relative to oocyte function and age-related fertility.<b>Abbreviations:</b> CCs: cumulus cells; COC: cumulus-oocyte complex; LC-MS: liquid chromatography-mass spectrometry; acetyl-CoA: acetyl-Coenzyme A; CoA: Coenzyme A.</p>","PeriodicalId":22184,"journal":{"name":"Systems Biology in Reproductive Medicine","volume":"68 1","pages":"36-43"},"PeriodicalIF":2.4,"publicationDate":"2022-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8821170/pdf/nihms-1766969.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10651339","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Investigating the role of EGF-CFC gene family in recurrent pregnancy loss through bioinformatics and molecular approaches. 利用生物信息学和分子方法研究EGF-CFC基因家族在复发性妊娠丢失中的作用。
IF 2.4 4区 医学 Q3 ANDROLOGY Pub Date : 2021-12-01 Epub Date: 2021-09-09 DOI: 10.1080/19396368.2021.1965673
João Matheus Bremm, Juliano André Boquett, Marcus Silva Michels, Thayne Woycinck Kowalski, Flávia Gobetti Gomes, Fernanda Sales Luiz Vianna, Maria Teresa Vieira Sanseverino, Lucas Rosa Fraga
ABSTRACT Recurrent pregnancy loss (RPL) is the most common reproductive failure, reaching 1-5% of women throughout their lives, and having unknown etiology in 50% of the cases. In humans, EGF-CFC1 (Epidermal Growth Factors & Cripto/FRL-1/Cryptic) gene family is composed by TDGF1 and CFC1, two developmental genes. The aim of this study was to investigate the role of EGF-CFC on RPL. To this, multiple approaches were performed; we conducted an expression analysis of TDGF1 and CFC1 using publicly available data from Gene Omnibus Expression (GEO), systems biology analyses and functional prediction; and a molecular analysis carried out in a case-control study. Our GEO analysis showed a decrease in TDGF1 expression in the endometrium (p=0.049) and CFC1 expression in placenta (p=0.015) of women with RPL. Network analysis, gene ontology and literature pointed to a strong connection between EGF-CFC1 gene family to pathways that play key roles during pregnancy, including TGF-β, c-Src/MAPK/AKT, Notch, TNFα, IFNγ and IL-6. A pathogenicity score developed for this gene family showed that the c.-14+1429T>C (rs3806702) variant in the TDGF1 and the p.Arg47Gln (rs201431919) variant in CFC1 gene would be the ones with the highest deleterious effect for RPL. In the case-control study, which involved 149 women with RPL and 159 controls, no statistical difference was observed in the allele and genotype distributions of the variants studied in the two groups. In this study, we performed extensive bioinformatics analysis for biomarker prioritization followed by experimental validation of proposed selected markers. Although there is no statistical difference in the frequencies of these variants between RPL and controls, the expression analysis results suggest that TDGF1 and CFC1 genes might play a role in RPL. In addition, systems biology analyzes raise the hypothesis that genes in other signaling pathways that may be related to RPL as good candidates for future studies. Abbreviations RPL: recurrent pregnancy loss; EGF-CFC1: Epidermal Growth Factors – Cripto/FRL-1; GEO: Gene Omnibus Expression; KEGG: Kyoto Encyclopedia of Genes and Genomes
复发性妊娠丢失(RPL)是最常见的生殖失败,占女性一生的1-5%,其中50%的病例病因不明。在人类中,EGF-CFC1 (Epidermal Growth Factors & crypto /FRL-1/Cryptic)基因家族由TDGF1和CFC1这两个发育基因组成。本研究的目的是探讨EGF-CFC在RPL中的作用。为此,采取了多种方法;我们利用基因综合表达(GEO)、系统生物学分析和功能预测的公开数据对TDGF1和CFC1进行了表达分析;在病例对照研究中进行了分子分析。我们的GEO分析显示,RPL患者子宫内膜中TDGF1表达(p=0.049)和胎盘中CFC1表达(p=0.015)均有所下降。网络分析、基因本体论和文献表明,EGF-CFC1基因家族与TGF-β、c-Src/MAPK/AKT、Notch、TNFα、IFNγ和IL-6等在妊娠过程中发挥关键作用的通路密切相关。对该基因家族的致病性评分显示,TDGF1基因中的C -14+1429T>C (rs3806702)变异和CFC1基因中的p.a g47gln (rs201431919)变异对RPL的危害最大。在病例对照研究中,涉及149名RPL妇女和159名对照组,在两组研究的变异的等位基因和基因型分布中没有观察到统计学差异。在这项研究中,我们对生物标志物的优先级进行了广泛的生物信息学分析,然后对所选标记进行了实验验证。虽然RPL和对照组之间这些变异的频率没有统计学差异,但表达分析结果表明TDGF1和CFC1基因可能在RPL中发挥作用。此外,系统生物学分析提出了其他信号通路中可能与RPL相关的基因作为未来研究的良好候选者的假设。RPL:复发性妊娠丢失;EGF-CFC1:表皮生长因子- crypto /FRL-1;GEO:基因综合表达;京都基因和基因组百科全书。
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引用次数: 0
Immunohistochemical examination of androgen receptor and estrogen receptor alpha expressions in obstructive and non-obstructive azoospermia. 梗阻性和非梗阻性无精子症患者雄激素受体和雌激素受体α表达的免疫组化检测。
IF 2.4 4区 医学 Q3 ANDROLOGY Pub Date : 2021-12-01 Epub Date: 2021-08-22 DOI: 10.1080/19396368.2021.1958094
Yurdun Kuyucu, Gülfidan Coşkun, Dilek Şaker, Özdem Karaoğlan, İbrahim Ferhat Ürünsak, Volkan İzol, İbrahim Atilla Arıdoğan, Şeyda Erdoğan, Hülya Özgür, Sait Polat

In this study, the expression of the androgen receptor (AR) and estrogen receptor alpha (ERα) in testicular tissue of male patients with obstructive azoospermia (OA) and non-obstructive azoospermia (NOA) were evaluated by immunohistochemistry. NOA (n = 23) and OA (n = 21) groups were created according to clinical and laboratory archival records. Testicular sperm extraction tissue sections were evaluated according to Johnsen's tubular biopsy scoring (JTBS) method. ERα and AR immunostaining results were evaluated semiquantitatively. Serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone, and estradiol were analyzed. Serum FSH and LH concentrations were greater, and testosterone concentrations were lower than the normal values in the NOA group, whereas the OA group revealed normal hormonal values. Serum estradiol concentrations in groups were in the normal range. JTBSs were significantly lower in the NOA group. Decreased AR expression and increased ERα expression were observed in the NOA group compared to the OA group. This suggests that ERα and AR are expressed in Sertoli cells, Leydig cells, and myoid cells and are required for normal testicular function. Decreased expression of the AR and increased expression of ERα in the testis may negatively affect spermatogenesis.Abbreviations: AR: androgen receptor; ER: estrogen receptor; ERα: estrogen receptor alpha; FSH: follicle-stimulating hormone; JTBS: Johnsen's tubular biopsy scoring; LH: luteinizing hormone; NOA: non-obstructive azoospermia; OA: obstructive azoospermia; TESE: testicular sperm extraction.

本研究采用免疫组织化学方法检测男性阻塞性无精子症(OA)和非阻塞性无精子症(NOA)患者睾丸组织中雄激素受体(AR)和雌激素受体α (ERα)的表达。根据临床和实验室档案记录创建NOA组(n = 23)和OA组(n = 21)。根据Johnsen管状活检评分法(JTBS)对睾丸取精组织切片进行评价。对ERα和AR免疫染色结果进行半定量评价。分析血清促卵泡激素(FSH)、黄体生成素(LH)、睾酮和雌二醇水平。NOA组血清FSH和LH浓度高于正常值,睾酮浓度低于正常值,而OA组的激素值正常。各组血清雌二醇浓度均在正常范围内。NOA组jtbs明显降低。与OA组相比,NOA组AR表达降低,ERα表达升高。这表明ERα和AR在支持细胞、间质细胞和肌样细胞中表达,是正常睾丸功能所必需的。睾丸中AR表达的减少和ERα表达的增加可能会对精子发生产生负面影响。缩写:AR:雄激素受体;ER:雌激素受体;ERα:雌激素受体α;促卵泡激素;JTBS:约翰森肾小管活检评分;LH:黄体生成素;NOA:非阻塞性无精子症;OA:阻塞性无精子症;睾丸精子提取。
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引用次数: 4
Production of bioactive recombinant ovine cysteine-rich secretory protein 1 in Escherichia coli. 重组羊富半胱氨酸分泌蛋白1在大肠杆菌中的生产。
IF 2.4 4区 医学 Q3 ANDROLOGY Pub Date : 2021-12-01 Epub Date: 2021-08-29 DOI: 10.1080/19396368.2021.1963012
Kalpana Jorasia, Rajani Kr Paul, N S Rathore, Pyare Lal, R Singh, Meenaxi Sareen

Ovine cysteine-rich secretory protein 1 (CRISP-1) is an acidic glycoprotein of epididymal origin under CRISP, antigen 5, pathogenesis-related protein 1 (CAP) super-family. The aim of the present study was the optimization of bacterial production and partial characterization of putative mature ovine CRISP-1 protein. The cDNA corresponding to T23 - C242 peptide fragment of ovine CRISP-1 protein was cloned into THE pET32b(+) expression vector using E. coli DH5α. Protein expression was carried out in E. coli BL21(DE3) by inducition with 1 mM IPTG at 37°C for 4 h. The recombinant protein was expressed as inclusion bodies and purified by Ni-NTA affinity chromatography using a pH gradient. Further purification of the protein was carried out by gel extraction following zinc sulfate negative staining. SDS-PAGE analysis of the purified recombinant CRISP-1 protein revealed a 43.8 kDa band. Bioactivity of the purified CRISP-1 protein was examined on sperm motility and capacitation. The recombinant ovine CRISP-1 protein at 5 µg/ml caused significant inhibition of sperm motility, and the activity was lost following heating the protein at 100°C for 5 min. The protein also demonstrated decapacitation activity, and at a concentration of 2 µg/ml, it caused a significant (P < 0.05) reduction in sperm capacitation. In conclusion, the thioredoxin-tagged ovine CRISP-1 protein was successfully produced in E. coli and purified in the soluble form by a combination of Ni-NTA affinity chromatography, gel purification, and dialysis. The recombinant protein exhibited both motility-inhibiting and decapacitating activities. Further study is needed to elucidate the mechanism of action and evaluate it's possible use in semen preservation.Abbreviations: CRISP-1: Cysteine-rich secretory protein-1; PCR: polymerase chain reaction; IPTG: isopropyl-β-D-thiogalactopyranoside; LB: Luria Bertani; SDS-PAGE: sodium dodecyl sulfate polyacrylamide gel electrophoresis; EDTA: ethylene diamine tetraacetic acid; Ni-NTA: Nickel nitrilotriacetic acid.

绵羊富含半胱氨酸分泌蛋白1 (CRISP-1)是一种起源于附睾的酸性糖蛋白,属于CRISP、抗原5、致病相关蛋白1 (CAP)超家族。本研究的目的是优化细菌生产和部分鉴定推定成熟的绵羊CRISP-1蛋白。利用大肠杆菌DH5α将羊CRISP-1蛋白T23 - C242肽片段cDNA克隆到The pET32b(+)表达载体上。在大肠杆菌BL21(DE3)中,用1 mM IPTG在37℃下诱导表达4 h。重组蛋白以包涵体形式表达,采用Ni-NTA亲和层析纯化。在硫酸锌阴性染色后,用凝胶萃取法进一步纯化蛋白。纯化的重组CRISP-1蛋白的SDS-PAGE分析显示有43.8 kDa的条带。纯化后的CRISP-1蛋白在精子运动和获能方面的生物活性进行了检测。5µg/ml的重组羊CRISP-1蛋白对精子活力有明显的抑制作用,在100℃下加热5 min后活性丧失。该蛋白也表现出失活活性,在2µg/ml的浓度下,该蛋白引起了显著的大肠杆菌(P . coli),并通过Ni-NTA亲和层析、凝胶纯化和透析相结合以可溶性形式纯化。重组蛋白具有运动抑制和失能活性。其作用机制有待进一步研究,并评价其在精液保存中的应用前景。缩写:CRISP-1:富含半胱氨酸分泌蛋白1;PCR:聚合酶链反应;IPTG:异丙酯-β-D-thiogalactopyranoside;LB: Luria Bertani;SDS-PAGE:十二烷基硫酸钠聚丙烯酰胺凝胶电泳;EDTA:乙二胺四乙酸;Ni-NTA:硝基三乙酸镍。
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引用次数: 0
Relationship between functional Nrf2 gene promoter polymorphism and sperm DNA damage in male infertility. 男性不育症Nrf2基因启动子多态性与精子DNA损伤的关系
IF 2.4 4区 医学 Q3 ANDROLOGY Pub Date : 2021-12-01 Epub Date: 2021-09-19 DOI: 10.1080/19396368.2021.1972359
O Sena Aydos, Yunus Yukselten, Dunya Aydos, Asuman Sunguroglu, Kaan Aydos

This study examines the association of the -617 C > A polymorphism in the Nrf2 gene (rs6721961) with male infertility in a Turkish population and determines its functional role in spermatogenesis in correlation with the impact of different levels of DNA damage on the genotypes. A total of 100 infertile men and 100 healthy fertile men were included in the study. Nrf2 genotyping was performed with the PCR-based restriction fragment length gene polymorphism (RFLP-PCR) analysis. According to our results, the Nrf2 CC, CA, and AA genotype distribution frequencies were 58.6%, 38.4%, and 3% in the control group, respectively, and 38%, 48%, and 14% in the infertile men, respectively. The AA genotype was significantly higher in the patient group. In smokers, a significant difference was found in progressive motility values between the genotypes (p = 0.001). Also, sperm progressive motility and concentration decreased significantly in those smokers with the AA genotype; smokers carrying this genotype were also 5.75 times more likely to have oligoasthenozoospermia than those with CC (p < 0.05). There was a significant relationship between the number of cases with high sperm-DNA damage when comparing the frequency of Nrf2 AA genotype carriers with the CC genotype 16.3% vs. 6.9%, respectively (p < 0.001). These results suggest the importance of the Nrf2 gene C > A (rs 6,721,961) polymorphism in the etiology of sperm DNA damage as a risk factor for male infertility. Smokers carrying the AA genotype are more likely to impair seminal parameters through antioxidant mechanisms.Abbreviations: Polymerase chain reaction (PCR)-based restriction fragment length gene polymorphism (RFLP-PCR); reactive oxygen species (ROS); deoxyribonucleic acid (DNA); catalases (CATs); superoxide dismutase (SOD); glutathione peroxidase (GPX); glutathione-S-transferase (GST); Nuclear factor erythroid 2 (NF-E2)-related factor 2 (Nrf2); basic leucine zipper (bZIP); antioxidant response element (ARE); World Health Organization (WHO);normospermia(NS);asthenozoospermia(AS);oligozoospermia(OS);oligoasthenozoospermia (OAS); follicle stimulating hormone (FSH); ultraviolet (UV); low-melting-point agarose (LMA); normal-melting-point agarose (NMA); arbitrary units (AU); total comet score (TCS); A one-way analysis of variance (ANOVA); standard deviation (SD); N-acetyltransferase (NAT2); small non-coding RNAs (ncRNAs); microRNAs (miRNA).

本研究探讨了土耳其人群中Nrf2基因(rs6721961) -617 C > A多态性与男性不育的关系,并确定了其在精子发生中的功能作用,以及不同水平的DNA损伤对基因型的影响。共有100名不育男性和100名健康的有生育能力的男性参与了这项研究。采用限制性内切片段长度基因多态性(RFLP-PCR)分析Nrf2基因分型。结果显示,Nrf2 CC、CA和AA基因型分布频率在对照组分别为58.6%、38.4%和3%,在不育男性中分别为38%、48%和14%。患者组AA基因型明显增高。在吸烟者中,基因型之间的渐进式运动值存在显著差异(p = 0.001)。AA基因型吸烟者精子进行性活力和精子浓度显著降低;在精子DNA损伤的病因学中,携带该基因型的吸烟者患少弱精子症的可能性是携带CC多态性的吸烟者的5.75倍(p A (rs 6,721,961))。携带AA基因型的吸烟者更有可能通过抗氧化机制损害精液参数。基于聚合酶链反应(PCR)的限制性片段长度基因多态性(RFLP-PCR);活性氧(ROS);脱氧核糖核酸;过氧化氢酶(猫);超氧化物歧化酶(SOD);谷胱甘肽过氧化物酶(GPX);glutathione-S-transferase(销售税);核因子红系2 (NF-E2)相关因子2 (Nrf2);碱性亮氨酸拉链(bZIP);抗氧化反应元件(ARE);世界卫生组织(WHO);无精子症(NS);无精子症(AS);少精子症(OS);少精子症(OAS);促卵泡激素(FSH);紫外线(UV);低熔点琼脂糖(LMA);正熔点琼脂糖;任意单位(AU);总彗星分数;单向方差分析(ANOVA);标准差(SD);N-acetyltransferase (NAT2);小非编码rna (ncRNAs);小分子核糖核酸(microrna的)。
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引用次数: 4
Levels of inter-alpha-trypsin inhibitor heavy chain H4 urinary polypeptide in gestational diabetes mellitus. 胰蛋白酶间抑制剂重链H4尿多肽在妊娠期糖尿病中的水平。
IF 2.4 4区 医学 Q3 ANDROLOGY Pub Date : 2021-12-01 Epub Date: 2021-10-04 DOI: 10.1080/19396368.2021.1977869
Zhiying Hu, Junlin Hou, Man Zhang

Gestational diabetes mellitus (GDM) can cause a variety of adverse maternal and fetal complications. The purpose of this study was to screen and identify the urinary polypeptides related to the severity of GDM and to analyze the correlation between urinary peptide levels and neonatal metabolic indices. A total of 31 normal pregnant women (N group) and 74 patients with GDM (GDM group) were randomly selected between February 2018 and August 2019. Patients with GDM were divided into two groups according to their fasting plasma glucose (FPG) levels. The urine samples were enriched using weak cation-exchange magnetic beads (MB-WCX), and eight different urine polypeptides were screened and analyzed. The peptide spectra were obtained using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). The urinary peptide signatures of the two groups were compared using the BioExplorer software. The difference analysis of the eight urinary polypeptides between the normal pregnant (N) group and GDM group showed that two polypeptides with mass-to- charge ratios (m/z) of 2175.7 and 2318.8, respectively, were significantly different between the two groups (P < 0.01). The m/z 2175.7 polypeptide was analyzed by liquid chromatography-tandem mass spectrometry (LC-MS), and the corresponding name of the molecule was inter-alpha-trypsin inhibitor heavy chain H4 (ITIH4). The changes in ITIH4 levels correlated with those in the neonatal metabolic indices. By establishing the Fisher discriminant function equation for the GDM group, the difference in sample distribution and mean value of the two groups could be observed directly.Abbreviations: GDM: gestational diabetes mellitus; FPG: fasting plasma glucose; MB-WCX: weak cation exchange magnetic beads; MALDI-TOF MS: matrix-assisted laser desorption ionization time-of-flight mass spectrometry; m/z: mass charge ratio; LC-MS: liquid chromatography-tandem mass spectrometry; glycosylated hemoglobin (HbA1c); PPG: postprandial plasma glucose; ITIH4: inter-alpha-trypsin inhibitor heavy chain H4; IR: insulin resistance; NFPG: neonatal fasting plasma glucose; NH: neonatal height; NW: neonatal weight; BMI: body mass index; RPL: recurrent pregnancy loss; OGTT: oral glucose tolerance test; ADA: American Diabetes Association; LIS: Laboratory Information System.

妊娠期糖尿病(GDM)可引起多种不良的母体和胎儿并发症。本研究旨在筛选和鉴定与GDM严重程度相关的尿多肽,并分析尿多肽水平与新生儿代谢指标的相关性。2018年2月至2019年8月,随机选择31名正常孕妇(N组)和74名GDM患者(GDM组)。根据空腹血糖(FPG)水平将GDM患者分为两组。用弱阳离子交换磁珠(MB-WCX)富集尿液样本,筛选并分析8种不同的尿液多肽。利用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)获得肽谱。使用BioExplorer软件比较两组的尿肽特征。正常妊娠(N)组与妊娠糖尿病(GDM)组8种尿多肽的差异分析显示,两组间质量电荷比(m/z)分别为2175.7和2318.8的2种多肽差异有统计学意义(P: GDM:妊娠糖尿病;FPG:空腹血糖;MB-WCX:弱阳离子交换磁珠;MALDI-TOF MS:基质辅助激光解吸电离飞行时间质谱法;M /z:质量电荷比;LC-MS:液相色谱-串联质谱;糖化血红蛋白(HbA1c);PPG:餐后血糖;ITIH4:胰蛋白酶间抑制剂重链H4;IR:胰岛素抵抗;NFPG:新生儿空腹血糖;NH:新生儿身高;NW:新生儿体重;BMI:身体质量指数;RPL:复发性流产;OGTT:口服葡萄糖耐量试验;ADA:美国糖尿病协会;实验室信息系统。
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引用次数: 0
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Systems Biology in Reproductive Medicine
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