Proteomics and polycystic ovary syndrome

Polycystic ovary syndrome (PCOS) is a complex endocrine disorder of heterogeneous etiology. Proteomics techniques have been used for elucidating the physiopathology of PCOS, yet the proteins identified so far were rarely the same across tissues and studies. The present review discusses the current challenges in the application of proteomics to the study of PCOS. A well-defined research design and an appropriate selection of study populations, samples and proteomic platforms are essential in clinical proteomics. Furthermore, the findings derived from proteomic approaches should be validated by complementary techniques, and the reproducibility of the results has ideally to be confirmed by different studies. Only when meeting these requirements, the proteins identified by proteomic techniques should be considered as candidates for future studies aiming to define specific molecular phenotypes of PCOS and their possible role in the metabolic and hormonal abnormalities characteristic of this syndrome.     

Nontargeted metabolomic analysis of skeletal muscle in a dehydroepiandrosterone‐induced mouse model of polycystic ovary syndrome

Polycystic ovary syndrome (PCOS) is the most common endocrinopathy and an important metabolic disorder in women of reproductive age. Insulin resistance (IR) is one of its most important clinical features in patients with PCOS. Androgen excess‐induced mitochondrial dysfunction contributes to skeletal muscle IR in dehydroepiandrosterone (DHEA)‐induced PCOS mice. The effect of androgen excess on the skeletal muscle, however, is incompletely characterized. A nontargeted metabolomics approach was thus applied to analyze the metabolites in skeletal muscle of DHEA‐induced PCOS mice. Data from metabolomic analysis revealed the significant changes in 32 metabolites and the marked impact of five metabolic pathways. ATP production was also found to be significantly reduced in skeletal muscle of DHEA mice. Combined with the quantification of type I and II myofibers and lipid measurement in the skeletal muscle of the mice, the results from the present study supported the role of mitochondrial impairment rather than lipid accumulation in the pathogenesis of skeletal muscle IR in DHEA‐induced PCOS mice. In summary, we show here for the first time the profile of the metabolites in the skeletal muscle of DHEA‐induced PCOS mice which exhibit IR. The work would help better understand the pathology of skeletal muscle IR in PCOS.     

多囊卵巢综合征患者血清VitD水平及与炎症因子之间的相关性研究

目的 了解多囊卵巢综合征（PCOS）患者血清中25-羟维生素D（25-OH-VitD3）水平情况,并探讨其与白细胞介素-6（IL-6）、肿瘤坏死因子（TNF-α）、转化生长因子（TGF-β）、单核细胞趋化蛋白-1（MCP-1）及C-反应蛋白（CRP）之间的相关性。方法 选择2017年8月—2018年12月来医院妇科门诊就诊并确诊为PCOS的患者共135例作为PCOS组,选择同期来医院体检的健康妇女共120例作为对照组,采用化学发光法检测血清中25-OH-VitD3水平,免疫比浊法检测CRP水平及采用酶联免疫吸附法（ELISA）检测IL-6、TNF-α及TGF-β水平,并对检测结果进行统计学分析。结果 PCOS组患者血清中25-OH-VitD3水平为31.87±6.29 nmol/L,明显低于对照组的65.08±13.76 nmol/L,差异有统计学意义（t=3.190,P<0.05）。PCOS组患者血清IL-6、TNF-α、TGF-β、MCP-1及CRP水平分别为23.71±11.23 ng/L、20.54±3.16 ng/L、76.54±8.03 ng/L、375.92±40.15 ng/L及13.48±3.10 mg/L,均明显高于对照组,差异均有统计学意义（t=9.065,8.091,10.178,13.052,7.059,Ps<0.05）。通过Pearson相关性分析,25-OH-VitD3水平与IL-6、TNF-α、TGF-β、MCP-1及CRP水平均呈明显的负相关（r= -0.562,-0.628,-0.780,-0.850,-0.505,Ps<0.05）。结论 25-OH-VitD3低表达水平可能通过其免疫调节及抑制炎症因子生成能力下降,引起各种炎症因子水平升高,导致PCOS的发病。因此,维生素D可能与PCOS病的发生和发展有密切关系。     

Serum ferritin as a candidate diagnostic biomarker of polycystic ovarian syndrome: a meta-analysis

Objective: In this study, we investigated about the potential of serum ferritin as a complementary diagnostic biomarker of polycystic ovarian syndrome (PCOS) by performing a meta-analysis of existing literature.

Materials and methods: Eleven studies written in English were retrieved up to 30 June 2018. Data were extracted from the selected studies by two of the authors and was subjected to statistical analysis. Levels of serum ferritin were compared between women with PCOS and controls using the standardized mean difference (SMD) and 95% confidence interval (CI). Subgroup analysis was also performed and stratified by ethnicity (Asians versus Caucasians).

Results: Overall post-outlier outcomes indicated that elevated serum ferritin is strongly associated with PCOS (SMD: 0.52; 95% CI: 0.40–0.64; P^A?=?10^?5). Subgroup analysis by ethnicity showed no significant difference between Asian and Caucasian population. Post-outlier receiving operations characteristics curve were plotted and showed that values for serum ferritin showed good potential in discriminating patients with and without PCOS (AUC?=?0.827, p?=?0.006).

Conclusion: Our findings suggest that high serum ferritin level is significantly associated with PCOS and its potential as a biomarker is evident in its high diagnostic accuracy. However, additional studies are needed to confirm our claims.     

microRNAs and long non‐coding RNAs as biomarkers for polycystic ovary syndrome

Polycystic ovary syndrome (PCOS) is known as the most common metabolic/endocrine disorder among women of reproductive age. Its complicated causality assessment and diagnostic emphasized the role of non‐coding regulatory RNAs as molecular biomarkers in studying, diagnosing and even as therapeutics of PCOS. This review discusses a comparative summary of research into microRNAs (miRNAs) and long non‐coding RNAs (lncRNAs) that are molecularly or statistically related to PCOS. We categorize the literature in terms of centering on either miRNAs or lncRNAs and discuss the combinatory studies and promising ideas as well. Additionally, we compare the pros and cons of the prominent research methodologies used for each of the abovementioned research themes and discuss how errors can be stopped from propagation by selecting correct methodologies for future research. Finally, it can be concluded that research into miRNAs and lncRNAs has the potential for identifying functional networks of regulation with multiple mRNAs (and hence, functional proteins). This new understanding may eventually afford clinicians to control the molecular course of the pathogenesis better. With further research, RNA (with statistical significance and present in the blood) may be used as biomarkers for the disease, and more possibilities for RNA therapy agents can be identified.     

Improvement of the folliculogenesis by transplantation of bone marrow mesenchymal stromal cells in mice with induced polycystic ovary syndrome

Background

Many studies have reported that inflammation and oxidative stress are involved in the pathogenesis of polycystic ovary syndrome (PCOS). Bone marrow mesenchymal stromal cells (BM-MSCs) have anti-oxidant and anti-inflammation properties. In this study, we investigate the beneficial effect of stem cell therapy on folliculogenesis in mice with induced PCOS

Programmed cell death 4: A novel player in the pathogenesis of polycystic ovary syndrome

Polycystic ovary syndrome (PCOS) is a pathological condition recognized by menstrual cycle irregularities, androgen excess, and polycystic ovarian morphology, affecting a significant proportion of women of childbearing age and accounting for the most prevalent cause of anovulatory sterility. In addition, PCOS is frequently accompanied by metabolic and endocrine disturbances such as obesity, dyslipidemia, insulin resistance, and hyperinsulinemia, indicating the multiplicity of mechanisms implicated in the progression of PCOS. However, the exact pathogenesis of PCOS is yet to be elucidated. Programmed cell death 4 (PDCD4) is a ubiquitously expressed protein that contributes to the regulation of various cellular processes, including gene expression, cell cycle progression, proliferation, and apoptosis. Despite some disparities concerning its exact cellular effects, PDCD4 is generally characterized as a protein that inhibits cell cycle progression and proliferation and instead drives the cell into apoptosis. The apoptosis of granulosa cells (GCs) is speculated to take a major part in the occurrence and progression of PCOS by ceasing antral follicle development and compromising oocyte competence. Given the possible involvement of GC apoptosis in the progression of PCOS, as well as the contribution of PDCD4 to the regulation of cell apoptosis and the development of metabolic diseases, the current review aimed to discuss whether or how PDCD4 can play a role in the pathogenesis of PCOS by affecting GC apoptosis.     

多囊卵巢综合征患者肠道菌群和生化免疫分子特征

【目的】比较多囊卵巢综合征(polycysticovariansyndrome,PCOS)患者与健康对照间肠道菌群谱结构特征差异,并探讨多囊卵巢综合征患者差异菌群与生化免疫指标之间的联系。【方法】选取初诊多囊卵巢综合征患者23例和健康女性对照23例,每例患者于初诊时留取粪便标本和血液标本,粪便标本提取DNA进行宏基因组测序,血清用于检测性激素和炎症因子。通过生物信息分析比较多囊卵巢综合征患者与健康对照间肠道菌群谱结构特征差异,以及与相关指标的联系。【结果】发现多囊卵巢综合征患者与健康对照的肠道菌群在门、属、种水平均有明显差异。两组间差异菌种为4个,普通拟杆菌(Bacteroides vulgatus)和弗格森埃希菌(Escherichia fergusonii)在PCOS组中的相对丰度高于对照组,并且这两种菌与PCOS的发病机制密切相关。凸腹真杆菌(Eubacteriumventriosum)和Subdoligranulum unclassified菌的相对丰度低于健康对照组。其中普通拟杆菌、凸腹真杆菌和Subdoligranulum unclassified菌3个菌种彼此间存在相关性...     

Microanalysis of N-linked oligosaccharides in a glycoprotein by capillary liquid chromatography/mass spectrometry and liquid chromatography/tandem mass spectrometry

We have studied rapid and simple sugar mapping using liquid chromatography/electrospray ionization mass spectrometry (LC/MS) equipped with a graphitized carbon column. The oligosaccharide mixture was separated on the basis of the sequence, branching structure, and linkage, and each oligosaccharide was characterized based on its molecular mass. In this study we demonstrated the usefulness of capillary LC/MS (CapLC/MS) and capillary liquid chromatography/tandem mass spectrometry (CapLC/MS/MS) as sensitive means for accomplishing the structural analysis of oligosaccharides in a low-abundance glycoprotein. The carbohydrate heterogeneity and molecular mass information of each oligosaccharide can be readily obtained from CapLC/MS of a small amount of glycoprotein. CapLC/MS/MS provided b-ion series, which is informative with regard to monosaccharide sequence. Exoglycosidase digestion followed by CapLC/MS elucidated a carbohydrate residue linkage. Using this method, we characterized N-linked oligosaccharides in hepatocyte growth factor produced in mouse myeloma NS0 cells as the complex-type bi-, tri-, and tetraantennary terminated with N-glycolylneuraminic acids and alpha-linked galactose residues. Sugar mapping with CapLC/MS and CapLC/MS/MS is useful for monitoring glycosylation patterns and for structural analysis of carbohydrates in a low-abundance glycoprotein and thus will become a powerful tool in biological, pharmaceutical, and clinical studies.     

Vitamin D3 affects mitochondrial biogenesis through mitogen-activated protein kinase in polycystic ovary syndrome mouse model

Polycystic ovarian syndrome (PCOS) is a disorder characterized by oligomenorrhea, anovulation, and hyperandrogenism. Altered mitochondrial biogenesis can result in hyperandrogenism. The goal of this study was to examine the effect of vitamin D3 on mitochondrial biogenesis of the granulosa cells in the PCOS-induced mouse model. Vitamin D3 applies its effect via the mitogen-activated pathway kinase-extracellular signal-regulated kinases (MAPK-ERK1/2) pathway. The PCOS mouse model was induced by the injection of dehydroepiandrosterone (DHEA). Isolated granulosa cells were subsequently treated with vitamin D3, MAPK activator, and MAPK inhibitor. Gene expression levels were measured using real-time polymerase chain reaction. MAPK proteins were investigated by western blot analysis. We also determined reactive oxygen species (ROS) levels with 2′, 7′-dichlorofluorescein diacetate. Mitochondrial membrane potential (mtMP) was also measured by TMJC1. Mitochondrial biogenesis (peroxisome proliferator-activated receptor gamma coactivator 1-α and nuclear respiratory factor), antioxidant (superoxide dismutase, glutathione peroxidase, and catalase), and antiapoptotic (B-cell lymphoma-2) genes were upregulated in the PCOS mice that treated with vitamin D3 compared with the PCOS mice without any treatment. Vitamin D3 and MAPK activator-treated groups also reduced ROS levels compared with the nontreated PCOS group. In summary, vitamin D3 and MAPK activator increased the levels of mitochondrial biogenesis, MAPK pathway, and mtMP markers, while concomitantly decreased ROS levels in granulosa cells of the PCOS-induced mice. This study suggests that vitamin D3 may improve mitochondrial biogenesis through stimulation of the MAPK pathway in cultured granulosa cells of DHEA-induced PCOS mice which yet to be investigated.     

多囊卵巢综合征患者阴道微生态变化及其与性激素水平的关系

目的探讨多囊卵巢综合征(PCOS)患者阴道微生态变化及其与性激素水平的关系。方法将2018年7月-2019年7月我院生殖健康与不孕症科收治的132例PCOS患者作为研究组,选择同期在我院进行妇科体检的86例健康女性作为对照组。16S rRNA法和定量PCR反应检测阴道分泌物微生态情况,放射免疫法测定血清中睾酮(T)、卵泡刺激素(FSH)、黄体生成素(LH)、雌二醇(E2)、孕酮(P)、催乳素(PRL)水平,分析微生态变化与性激素水平的关系。结果研究组患者阴道分泌物清洁度Ⅳ度检出率,滴虫、假丝酵母菌及线索细胞阳性检出率,阴道微生态失调率,血清T、LH、E2及PRL水平均高于对照组(P0.05),研究组患者血清FSH、P水平低于对照组(P0.05)。阴道微生态失调患者血清中E2水平高于阴道微生态正常患者,P水平低于阴道微生态正常患者(P0.05)。结论 PCOS患者存在阴道微生态失衡,阴道微生态的变化与血清性激素水平存在相关性,考虑激素水平的变化可能参与阴道微生态失调的发生。     

Pathophysiological changes in experimental polycystic ovary syndrome in female albino rats: Using either hemin or L-arginine

Polycystic ovary syndrome (PCOS), one of the important endocrine disorders affecting females in the reproductive age, is caused mainly by an abnormal oxidation status that subsequently causes inflammatory conditions. Thus, this study aims to examine the possible individual prophylactic effects of gasotransmitters, hemin, or L-arginine in letrozole-induced PCOS. Fifty adult female albino rats were used and separated into a control group, which received the vehicle; a letrozole-induced PCOS group (L), which received letrozole orally at a dose level of 1 mg/kg for 21 days; a letrozole+hemin (L+H) group, which received letrozole plus hemin at a dose level of 25 mg/kg injected IP twice per week for 21 days; and a letrozole+L-arginine (L+A) group, which received letrozole plus L-arginine at a dose level of 200 mg/kg orally for 21 days. During PCO induction, the body weight and Lee index were measured. Serum glucose, insulin, lipid profile, gonadotrophic hormones, testosterone, estrogen, and tumor necrosis factor alpha were assayed, while ovarian tissues were analyzed to measure the oxidative state and histopathological changes. Our results proved that either hemin or L-arginine administration could improve the oxidative state, the inflammatory reaction, the hormonal imbalance, and the metabolic disturbances in PCO rats, which was confirmed by a histopathological examination of the rats’ ovaries. In conclusion, either hemin or L-arginine had protective effects against PCOS with better pathophysiological changes with hemin.     

N-acetyl cysteine reduces oxidative toxicity,apoptosis, and calcium entry through TRPV1 channels in the neutrophils of patients with polycystic ovary syndrome

Abstract

Polycystic ovary syndrome (PCOS) is a common inflammatory and oxidant disease with an uncertain pathogenesis. N-acetyl cysteine (NAC) decreases oxidative stress, intracellular free calcium ion [Ca²⁺]_i, and apoptosis levels in human neutrophil. We aimed to investigate the effects of NAC on apoptosis, oxidative stress, and Ca²⁺ entry through transient receptor potential vanilloid 1 (TRPV1) and TRP melastatin 2 (TRPM2) channels in neutrophils from patients with PCOS. Neutrophils isolated from PCOS group were investigated in three settings: (1) after incubation with TRPV1 channel blocker capsazepine or TRPM2 channel blocker 2-aminoethyl diphenylborinate (2-APB), (2) after supplementation with NAC (for 6 weeks), and (3) with combination (capsazepine + 2-APB + NAC) exposure. The neutrophils in TRPM2 and TRPV1 experiments were stimulated by N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP; 1 μM) and capsaicin (10 μM) as concentration agonists, respectively. Neutrophil lipid peroxidation and capsaicin-induced increase in [Ca²⁺]_i concentrations were reduced by capsazepine and NAC treatments. However, the [Ca²⁺]_i concentration did not change by fMLP stimulation. Neutrophil lipid peroxidation, apoptosis, caspase-3, caspase-9, cytosolic reactive oxygen species production, and mitochondrial membrane depolarization values were decreased by NAC treatment although neutrophil glutathione peroxidase and reduced glutathione levels were increased by the NAC treatment. Serum lipid peroxidation, luteinizing hormone, testosterone, insulin, interleukin-1 beta, and homocysteine levels were decreased by NAC treatment although serum vitamin A, beta-carotene, vitamin E, and total antioxidant status were increased by the NAC treatment. In conclusion, NAC reduced oxidative stress, apoptosis, cytokine levels, and Ca²⁺ entry through TRPV1 channel, which provide supportive evidence that oxidative stress and TRPV1 channel plays a key role in etiology of PCOS.     

Alterations of polyunsaturated fatty acid metabolism in ovarian tissues of polycystic ovary syndrome rats

The metabolism of polyunsaturated fatty acids (PUFAs) remains poorly characterized in ovarian tissues of patients with polycystic ovary syndrome (PCOS). This study aimed to explore alterations in the levels of PUFAs and their metabolites in serum and ovarian tissues in a PCOS rat model treated with a high‐fat diet and andronate. Levels of PUFAs and their metabolites were measured using gas/liquid chromatography‐mass spectrometry after the establishment of a PCOS rat model. Only 3 kinds of PUFAs [linoleic acid, arachidonic acid (AA) and docosahexaenoic acid] were detected in both the circulation and ovarian tissues of the rats, and their concentrations were lower in ovarian tissues than in serum. Moreover, significant differences in the ovarian levels of AA were observed between control, high‐fat diet‐fed and PCOS rats. The levels of prostaglandins, AA metabolites via the cyclooxygenase (COX) pathway, in ovarian tissues of the PCOS group were significantly increased compared to those in the controls. Further studies on the mechanism underlying this phenomenon showed a correlation between decreased expression of phosphorylated cytosolic phospholipase A2 (p‐cPLA2) and increased mRNA and protein expression of COX2, potentially leading to a deeper understanding of altered AA and prostaglandin levels in ovarian tissues of PCOS rats.     

多囊卵巢综合征患者与健康人群肠道菌群差异性分析

目的 比较多囊卵巢综合征(PCOS)患者与健康人群间肠道菌群的差异,为后续研究提供参考.方法 采用16SrDNA扩增子测序法对30例PCOS患者(PCOS组)和20例健康人(健康组)粪便标本中菌群结构进行分析,并比较两组之间的区别.结果 与健康组相比,PCOS组患者肠道菌群α多样性降低.PCOS组患者肠道厚壁菌门(Fi...     

Effects of serum irisin,neuregulin 4, and weight management on obese adolescent girls with polycystic ovary syndrome

The study is aimed at investigating the association of serum irisin, neuregulin 4 (NRG4), and anti-müllerian hormone (AMH) with adolescent obesity with polycystic ovary syndrome (PCOS) and the efficacy of weight management interventions. Serum levels of irisin, NRG4, AMH, sex steroid hormone, body mass index (BMI), serum insulin, and C-peptide were measured in 52 obese adolescent girls with PCOS (PCOS group) and 43 obese adolescent girls without PCOS (non-PCOS group). The levels of AMH, NRG4, serum irisin, sex steroid hormones, BMI, serum insulin, and C-peptide were evaluated in obese PCOS girls before and after one year weight management. The levels of AMH, serum insulin, NRG4, and total testosterone of PCOS group were significantly higher than those of non-PCOS group. On the contrary, serum irisin and serum C-peptide in PCOS group were significantly lower than that in non-PCOS group. The levels of fat mass, percent body fat, total testosterone, AMH, NRG4, and serum insulin in the obese girls with PCOS showed significant decreases compared with before weight management intervention. On the contrary, after one year of body weight management intervention, serum irisin and serum C-peptide was significantly increased. Adolescent obesity complicated with PCOS is significantly associated with glucose and lipid metabolism and sex steroid hormone disorders, but the exact pathophysiological and clinical features are highly variable. Weight management intervention can significantly improve the clinical symptoms and hematological indicators, serum irisin and NRG4 can be used as two essential biomarkers for evaluating weight management.