Disorders Of Male Reproductive System Research Articles

An RNA-seq transcriptome analysis for investigating the anti-lung cancer activity of medicinal Cuscuta chinensis Lam plant.

Cuscuta chinensis Lam. is a traditional medicinal herb used to treat female sterility and male reproductive system disorders. However, the anti-lung cancer properties of Cuscuta chinensis Lam. and possible molecular mechanisms have yet to be explored. Thus, the study’s main purpose was to evaluate in vitro and in vivo anti-lung cancer properties of C. chinensis water extract (CLW) in human lung adenocarcinomas and the underlying molecular mechanism involved. Our results demonstrated that CLW caused a significant inhibition of cell viability and induced G1 cycle arrest in lung cancer cells. Furthermore, RNA-seq transcriptome analysis revealed 602 common genes with a significant expression in A549 and H1650 cells under CLW treatment. Functional enrichment analysis suggested that these common genes regulated by CLW mainly involve lung cancer cell proliferation, metastases and apoptosis processes. In addition, forty-six common genes (> 2-fold change) regulated by CLW in A549 and H1650 cells were selected for further validation. In vitro quantitative real-time PCR results confirmed that twelve genes were up-regulated, and four genes were down-regulated in A549 and H1650 cells. The in vivo experiment demonstrated CLW could significantly decrease tumour volume and tumour weight of mice compared with the control group. Moreover, in vivo quantitative real-time PCR results revealed that C11orf96, FGFBP1, FOSB and NPTX1 genes were up-regulated and EGR1, GBP4 and MAP2K6 genes were down-regulated in tumour tissues compared with the control group. These data strongly suggest that CLW could be developed as an efficacious drug for lung cancer treatment.

Male infertility associated with oxidative stress of spermatozoa: pathogenesis and therapeutic approach

Infertility is defined as not being to get pregnant for a couple after at least a year of regular sexual life. The condition has become a global health issue affecting about 187 million couples worldwide, and about half of the cases are associated with male factors. There are several external and internal factor infertility. Oxidative stress, which is a common cause of several conditions associated with male infertility is one of the main factors. High levels of reactive oxygen species (ROS) decrease sperm quality through reducing sperm motility and increasing damage to DNA, protein, and lipids. If DNA strand breaks are not repaired, a cell undergoes apoptosis, programmed cell death. The combination antioxidants are considered effective in improving male fertility parameters due to synergistic effects of various components. The mechanism of action of most of them is directed toward reducing ROS concentration, thereby improving sperm quality. In addition, this review considers other natural molecules - myo-inositol and D-chiro-inositol, which improve sperm quality. They are involved in many signalling mechanisms in spermatozoa that regulate cytoplasmic calcium levels, mitochondrial capacity and function. On the other hand, D-chiro-inositol is involved in the inhibition of steroidogenic aromatase enzymes, thereby increasing testosterone levels. In this review, we analyse the processes associated with oxidative stress in male fertility and the mechanisms of action of various molecules, as well as consider treatment options for management of various male reproductive system disorders.

Exosomes as Potential Biomarkers for Erectile Dysfunction, Varicocele, and Testicular Injury.

Optimal male reproductive health is dependent upon critical mediators of cell-cell communication: exosomes or extracellular vesicles. These vesicles are nano-sized particles released into a variety of bodily fluids, such as blood and semen. Exosomes are highly stable and can carry genetic and other molecules, including DNA, RNA, and proteins, which provide information about their origin cells. To identify exosomes as potential biomarkers or therapeutic mediators in male sexual and reproductive disorders like erectile dysfunction (ED), varicocele, and testicular injury. A PubMed search was performed to highlight all articles available relating to exosomes and extracellular vesicles in the pathogenesis of different male sexual and reproductive disorders, and their importance in clinical use as both diagnostic markers and potential therapeutic mediators. Various male reproductive system disorders, such as ED, varicocele, and testicular injury, are linked to increased or decreased levels of exosomes. Exosomes have a higher number of molecules such as DNA, RNA, and proteins, which can give a more precise and comprehensive result when compared to other biomarkers. Exosomes can be considered as plausible diagnostic biomarkers for male sexual and reproductive diseases, with considerable advantages over other diagnostic procedures such as invasive tissue biopsy. Exosomes can carry cargo such certain drugs and therapeutic molecules making them a promising therapeutic approach. Several studies have begun to test treating various male sexual reproductive disorders with exosomes. Exosomes deliver many components that can regulate gene expression and target signaling pathways. Understanding how extracellular vesicles can be utilized as biomarkers in diagnosing men, particularly those with idiopathic erectile dysfunction, will not only aid in diagnosis but also help with making therapeutic targets. Khodamoradi K, Golan R, Dullea A, et al. Exosomes as Potential Biomarkers for Erectile Dysfunction, Varicocele, and Testicular Injury. Sex Med Rev 2022;10:311-322.

Epigenetic Regulation of TET1-SP1 During Spermatogonia Self-Renewal and Proliferation.

Spermatogonia are the source of spermatogenic waves. Abnormal spermatogonia can cause ab-normal spermatogenic waves, which manifest as spermatogenic disorders such as oligospermia, hypospermia, and azoospermia. Among them, the self-renewal of spermatogonia serves as the basis for maintaining the process of spermatogenesis, and the closely regulated balance between self-renewal and differentiation of spermatogonia can maintain the continuous production of spermatozoa. Tet methylcytosine dioxygenase 1(TET1) is an important epitope modifying enzyme that catalyzes the conversion of 5-methylcytosine (5-mC) to 5-hydroxymethylcytosine (5-hmC), thereby causing the methylation of specific genes site hydroxylation, enabling the DNA de-methylation process, and regulating gene expression. However, the hydroxymethylation sites at which TET1 acts specifically and the mechanisms of interaction affecting key differential genes are not clear. In the present study, we provide evidence that the expression of PLZF, a marker gene for spermatogonia self-renewal, was significantly elevated in the TET1 overexpression group, while the expression of PCNA, a proliferation-related marker gene, was also elevated at the mRNA level. Significant differential expression of SP1 was found by sequencing. SP1 expression was increased at both mRNA level and protein level after TET1 overexpression, while differential gene DAXX expression was downregulated at protein level, while the expression of its reciprocal protein P53 was upregulated. In conclusion, our results suggest that TET1 overexpression causes changes in the expression of SP1, DAXX and other genes, and that there is a certain antagonistic effect between SP1 and DAXX, which eventually reaches a dynamic balance to maintain the self-renewal state of spermatogonia for sustained sperm production. These findings may contribute to the understanding of male reproductive system disorders.

Weighted gene coexpression network analysis reveals ESR1, FLNA and Furin as hub genes for DEHP-induced prepubertal testicular injury

Di-(2-ethylhexyl) phthalate (DEHP) is an environmental endocrine disruptor that accumulates in organisms in various ways and induces male reproductive system disorders. In this study, we established a testicular injury model by gavage with different concentrations of DEHP. The testes were then collected for RNA sequencing (RNA-seq), and the results were analyzed by bioinformatics and verified by experiments. Our research results show that different concentrations of DEHP interfere with testicular development differently. Weighted gene coexpression network analysis (WGCNA) generated sixteen modules and identified the turquoise module as key. Then, estrogen receptor 1 (ESR1), filamin A (Flna) and Furin were identified as hub genes. qPCR and immunohistochemistry results revealed that all three hub genes were upregulated. We detected the locations of these genes by immunohistochemistry. ESR1 was mainly located in Leydig cells; Flna immunostaining is observed in the Leydig and some germ cells and Furin staining was seen in almost all types of testicular cells. Furthermore, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses showed enrichment mainly in MAPK signaling pathways, p53 signaling pathways, HIF-1 signaling pathways, protein processing in the endoplasmic reticulum, apoptosis, the cell cycle, RNA degradation, etc. This is the first study using WGCNA to investigate the mechanism of DEHP-induced injury in the prepubertal testis, providing new research angles to further understand the mechanism of DEHP-induced injury in the prepubertal testis.

Potential therapeutic effects of Ulva lactuca water fraction on monosodium glutamate-induced testicular and prostatic tissue damage in rats.

Male infertility is considered one of the most critical health problems that are expected to expand worldwide. Ulva lactuca is a species of green seaweeds which is known to be a rich source of many important nutrients. Accordingly, this study is designated to investigate the therapeutic role of Ulva lactuca water fraction (UL) against monosodium glutamate (MSG)-induced male reproductive system disorders in male rats. Ulva lactuca methanolic crude extract was prepared firstly, and then water-dissolved compounds of this crude methanolic extract were separated. Ulva lactuca water fraction active phenolic compounds were determined using high-performance liquid chromatography (HPLC). Thirty-two male rats were divided equally into four groups; male infertility was induced in sixteen experimental animals by MSG at dose of 15mg/Kg for 45days. Eight infertile animals were treated with 100mg/Kg of Ulva lactuca water fraction for 30days. The rest of the animals were divided into two control groups; one control group (eight animals) was used to study the effect of UL on healthy rats at dose of 100mg/Kg for 30days and healthy control group (eight animals). Semen quality parameters (concentration and motility ratio), serum testosterone, prostate-specific antigen (PSA), and phosphatases were estimated by using standard protocols. Moreover, prooxidants and endogenous antioxidant enzymes were measured in prostate and testis homogenates. In addition, relative expression of pro-inflammatory genes (inducible nitric oxide synthase (i-NOS), cyclooxygenase-2 (COX-2), tumor necrosis factor-α alpha (TNF-α), and tumor protein (P53)) were assessed in testicular and prostatic tissues. Finally, histological alterations were measured by hematoxylin and eosin (H&E) stain. Results revealed that Ulva lactuca water fraction contains active phenolic constituents responsible for its antioxidant bioactivity. Oral administration of MSG significantly induced histological alterations. Oxidative stress was observed with elevated levels of nitric oxide (NO), thiobarbituric acid reactive substances (TBARS), and xanthine oxidase (XO) activity in both testis and prostate tissues. MSG adversely affected prostate function via elevation of PSA, prostatic acid phosphatases (PAPs), and total acid phosphatases (TAPs). In addition, it upregulated pro-inflammatory genes in testis and prostate tissues. Meanwhile, MSG reduced serum testosterone, semen quality, and antioxidant enzyme activities (glutathione peroxidase (GPx), glutathione-s-transferase (GST), and superoxide dismutase (SOD)). Treatment with UL notably ameliorated the state of oxidative stress and downregulated the expression of pro-inflammatory gene markers. This study highlighted the potential efficacy of Ulva lactuca water fraction on MSG-induced male infertility in rats. Therapeutic effect of UL on oxidative stress and inflammation induced by MSG in testicular and prostatic tissues.

Ferulago angulata extract ameliorates epididymal sperm toxicity in mice induced by lead and diazinon.

The potential toxicity that results from environmental xenobiotics is not completely known. Increasing levels of heavy metals and the use of organophosphate pesticides (OPs) and their co-existence in the environment could be associated with an increasing incidence of male reproductive system disorders in humans and animals. Ferulago angulata is a dietary source of phenolic compounds with reported health benefits. This study was conducted to investigate whether an extract of Ferulago angulata could protect adult male NMRI mice against reproductive toxicity induced by lead acetate (PbAc), diazinon (DZN), or PbAc+DZN. Adult male NMRI mice were exposed to either 0.5% PbAc in drinking water, DZN (3mg/kg/day, intraperitoneal [i.p.] injection), or PbAc+DZN in the presence or absence of 400mg/kg/day Ferulago angulata hydroalcoholic extract (FAE) that was administered via gavage for 6weeks. Chronic exposure to PbAc, DZN, and PbAc+DZN decreased sperm quality, sperm chromatin maturity and integrity, increased oxidative stress and lipid peroxidation, and could reduce male fertility indices. Co-administration of FAE could reduce these negative effects. The Ferulago angulata extract should be considered as a useful natural extract for the treatment of male infertility, especially in males exposed to conditions which induce reproductive toxicity.

Sodium Arsenic Alters the Gene Expression of some Steroidogenic Genes in TM3 Leydig Cell

Arsenic is a broad-spectrum environmental contaminant with mutagenic, teratogenic and carcinogenic effects. Due to its widespread distribution in nature, drinking water is the most common source of arsenic exposure for the general population. In this study, we aimed to determine the effect of sodium arsenite on the viability and expression profile of steroidogenic genes in TM3 Leydig cells, responsible for testicular steroidogenesis. The TM3 Leydig cells were treated with sodium arsenic (384,8 µM or 7,6 mM) for 24 hours with LH (Luteinizing hormone) stimulation. The MTT assay was used for measuring cell viability, the expression level of key genes of the steroidogenesis was evaluated using RT-qPCR.The MTT assay showed that cell viability was decreased dose-dependently. RT-qPCR demonstrated that the expression level of CYP11A1, CYP17A1 were decreased as compared to the untreated control while StAR gene expression was found to be surprisingly high in the cell exposed to high-dose arsenic (p<0.05). The expression profile of two dehydrogenase; 17β-HSD and 3β-HSD was significantly reduced in high dose arsenic treated-group (p<0.05); but however, no statistical significance was found in low-dose. The RT-qPCR also showed that the expression SF-1 (Steroidogenic factor-1), a key regulator of adrenal and reproductive development, was significantly decreased in both low-dose and high-dose (p<0.05). Arsenic toxicity in Leydig cell leads to cell viability loss and cause a perturbation in key steroidogenic genes, reflecting the possible role of arsenic in infertility and male reproductive system disorders.

Exploring disease-specific methylated CpGs in human male genital abnormalities by using methylated-site display-amplified fragment length polymorphism (MSD-AFLP).

The incidence of male reproductive system disorders, especially hypospadias, has been increasing in developed countries since the latter half of the 20th century. Endocrine-disrupting chemicals from the environment are considered to be involved in hypospadias onset through epigenetic alterations. This pilot study aimed to explore disease-specific methylated CpGs in human patient samples using the methylated-site display-amplified fragment length polymorphism (MSD-AFLP) technique developed by our research group [1]. We compared clinical samples from hypospadias and phimosis patients. Foreskin and blood samples were collected from one- to two-year-old patients with hypospadias (N = 3) and phimosis (N = 3) during surgical treatment. MSD-AFLP analysis showed significantly decreased CpG-methylation levels of genes such as MYH11 and increased CpG-methylation levels of genes such as PLA2G15 in hypospadias patients. Hierarchical clustering analysis showed that genes with significantly altered CpG levels were more markedly altered in DNA from blood than from foreskin. Because of the small number of samples, further investigation is necessary to elucidate the association between variations in CpG levels in foreskin and blood DNA and male genital abnormalities. However, our MSD-AFLP method appears to be a useful tool for exploring disease-specific methylated-CpGs in human epidemiological studies.

Expression of Xenobiotic Biomarkers CYP1 Family in Preputial Tissue of Patients with Hypospadias and Phimosis and Its Association with DNA Methylation Level of SRD5A2 Minimal Promoter

Several epidemiological studies have suggested that the incidence of male reproductive organ malformations, including hypospadias or cryptorchidism, has increased due to fetal-stage exposure to environmental pollutants. However, the association of chemical exposure with the expression of target regulatory genes in the tissues of patients has not yet been reported. Because experimental approaches or clinical trials in human studies are limited, especially those using fetal and/or infants, it is difficult to obtain clear physiological evidence of mechanisms underlying male reproductive malformations. Thus, the lack of physiological evidence makes this issue controversial. We analyzed preputial tissues from patients with hypospadias (n = 23) and phimosis (n = 16). The atypical CYP1 family genes, CYP1A1 and CYP1B1, are potential biomarkers of environmental chemical exposure. We then compared the expression levels of CYP1A1 and CYP1B1 between hypospadias and phimosis samples by quantitative RT-PCR analysis. The mRNA expression levels of SRD5A2 and AR also were measured, because the androgen-related genes involved in the onset of disorders of male reproductive system. A significantly higher CYP1B1 expression level and a lower AR expression level were observed in the hypospadias groups than in the phimosis group. Positive correlations (P < 0.001) between the mRNA expression levels of the CYP1 family and SRD5A2 were found in patients with hypospadias but not in those with phimosis. Moreover, the methylation levels of the four genes were determined by bisulfite genomic sequencing. Although the SRD5A2 promoter region showed moderate methylation, no methylation was detected in CYP1A1, CYP1B1, or AR. There was no significant difference in SRD5A2 promoter methylation level between hypospadias and phimosis patients. Negative correlations were found between the methylation level of SRD5A2, especially at the − 221 Sp1 site, and the CYP1 family mRNA expression levels (CYP1A1, p = 0.002; CYP1B1, p = 0.007) in hypospadias patients, but not in phimosis patients. The significant positive association of mRNA expression level and the negative association of methylation level of the SRD5A2 gene with the mRNA expression levels of CYP1 family genes in the preputial tissue seem to indicate the chemical exposure of patients with hypospadias.

Ameliorative effects of Achillea millefolium inflorescences alcoholic extract against nicotine-induced reproductive failure in rat

BackgroundNicotine (Nic) is a major risk factor in the development of functional disorders of male reproductive system. Achillea millefolium; is highly regarded for medicinal activities, due to its antioxidant and anti-inflammatory properties. This study was carried out to evaluate whether Achillea millefolium (Achm) inflorescences alcoholic extract could serve as a protective agent in male reproductive male failures during Nic exposure in a rat model. MethodsTwenty-five adult male Wistar rats were categorized into the five groups. Tests 1 and 3 groups were received Nic at dose levels of 0.2 and 0.4mg/kg BW/day, respectively by IP injection. Tests 2 and 4 groups were received Nic at the same doses along with Achm at dose level of 120mg/kg BW/day. The study period took forty-eight days for all experimental groups. ResultsNic groups showed significant decreases in tubule differentiation index (TDI), sperm count, motility, stereological parameters and an increase in dead and abnormal sperms. Moreover, the reduction in total antioxidant capacity (TAC), superoxide dismutase (SOD) activity, serum levels of FSH, LH and testosterone, along with increased serum concentration of LDH were observed in the Nic groups. Total nitrite and malondealdehyde levels increased and total thiol molecules (TTM) levels decreased in testicular tissue in the Nic groups. Notably, Achm co-administration caused a contemporary recovery in above-mentioned parameters. ConclusionNic exerts major toxicity in testicular tissue and causes damages in several ways including, oxidative stress, whilst Achm imposes protective effect against Nic-induced reproductive failure, which may attribute to its antioxidant capacity.

Evaluation of minerals in the seminal plasma of azoospermic semen

Background: Several studies have reported that mineral elements are involved in the performance and maintenance of male fertility.Aim: The aim of present study was to evaluate the mineral elements such as sodium, potassium, calcium, magnesium, phosphorus, chlorine and iron found in the seminal plasma of men was examined at the Pasteur Institute of Ce DIvoire to determine new biochemical markers in azoospermia.Material and Methods: Analysis of seminal fluid was carried out with 30 samples of normozoospermic semen and 30 samples of azoospermic semen. The assay of calcium, phosphorus, magnesium and Iron was done using the automated Hitachi Roche Cobas c 311. The evaluation of the concentration level of chlorine, sodium and potassium in the seminal plasma was performed using the automated 9180 Roche Electrolyte Analyzer. The data collected were analyzed using the software Graph Pad Prism 5. P values less than 0.05 was considered the level of significance.Results: The results of this study showed that the correlation test performed between concentrations level of minerals in the seminal plasma of normozoospermic and azoospermic semen showed no significant difference statistically with P > 0.05.Conclusion: The evaluation of the concentration level of mineral elements such as sodium, potassium, calcium, magnesium, phosphorus, chlorine and iron in the seminal plasma of azoospermic and normozoospermic semen showed that the concentration level of these mineral elements has no diagnostic value in the male reproductive system disorders, including azoospermia.

“We don’t know what we need to learn”: Medical student perceptions of preparedness for practice in men’s health

Introduction: In response to poorer health status and lower use of healthcare services by men compared to women, the Australian National Male Health Policy (2010) includes a commitment to better workforce training. We present a formative evaluation assessing medical students’ preparedness and learning needs in relation to the care of men in order to inform the development of a commensurate curriculum framework. Method : A 4-item Likert scale survey was administered to final-year students from one Australian medical school (n=170; 64% female, 36% male; 74% response). The survey assessed coverage of male health topics in the curriculum and preparedness for men’s health practice. Additional focus groups with 13 students from four Australian medical schools aimed to provide context for the survey data and better understanding of learning needs. Results: Overall, 65% of students (67% male, 64% female) reported no or brief coverage of men’s health in the curriculum; 20% of students (14% male, 23% female) felt minimally prepared in men’s health practice. Students’ perceived learning needs mostly related to male reproductive system disorders. Focus group data highlighted a difference between the content/opportunities that exist in men’s health education compared with women’s health, stating that men’s health education tends to be “scattered” variably through the medical course and that student or educators’ personal interest tended to drive men’s health education. In addition, unlike opportunities to undertake sensitive examinations on females, practice opportunities for male genital and prostate examination were limited, with some reporting no coverage in their medical course. Conclusion : From a student perspective, there is a need to enhance men’s health education in Australian medical schools by incorporating clinically relevant men’s health across curricula using a defined framework.

Seminal plasma as a diagnostic fluid for male reproductive system disorders

Molecular biomarkers hold promise to advance the noninvasive diagnosis of male reproductive system disorders and facilitate the identification and management of these conditions through screening, early diagnosis and more accurate prognosis. Seminal plasma has great potential as a proximal fluid for protein biomarker discovery and as a clinical sample for noninvasive diagnostics. The seminal plasma proteome contains thousands of proteins and includes a large number of tissue-specific proteins that might accurately indicate a pathological process in the tissue of origin. Potential protein biomarkers for male reproductive system disorders are more abundant in seminal plasma than in blood serum or urine, and, therefore, are more easily identified and quantified in semen by mass spectrometry and other techniques. These methods have enabled elaboration of the composition of the seminal plasma proteome and the tissue specificity of seminal plasma proteins. Strategies have been developed to discover protein biomarkers in seminal plasma through integrated 'omics' approaches. Biomarkers of male infertility and prostate cancer are now emerging, and it is evident that seminal plasma has the potential to complement other diagnostic tools available in urology clinics.

Verification of Male Infertility Biomarkers in Seminal Plasma by Multiplex Selected Reaction Monitoring Assay

Seminal plasma is a promising biological fluid to use for noninvasive clinical diagnostics of male reproductive system disorders. To verify a list of prospective male infertility biomarkers, we developed a multiplex selected reaction monitoring assay and measured the relative abundance of 31 proteins in 30 seminal plasma samples from normal, nonobstructive azoospermia and post-vasectomy individuals. Median levels of some proteins were decreased by more than 100-fold in nonobstructive azoospermia or post-vasectomy samples, in comparison with normal samples. To follow up the most promising candidates and measure their concentrations in seminal plasma, heavy isotope-labeled internal standards were synthesized and used to reanalyze 20 proteins in the same set of samples. Concentrations of candidate proteins in normal seminal plasma were found in the range 0.1-1000 μg/ml but were significantly decreased in nonobstructive azoospermia and post-vasectomy. These data allowed us to select, for the first time, biomarkers to discriminate between normal, nonobstructive azoospermia, and post-vasectomy (simulated obstructive azoospermia) seminal plasma samples. Some testis-specific proteins (LDHC, TEX101, and SPAG11B) performed with absolute or nearly absolute specificities and sensitivities. Cell-specific classification of protein expression indicated that Sertoli or germ cell dysfunction, but not Leydig cell dysfunction, was observed in nonobstructive azoospermia seminal plasma. The proposed panel of biomarkers, pending further validation, could lead to a clinical assay that can eliminate the need for testicular biopsy to diagnose the category of male infertility, thus providing significant benefits to patients as well as decreased costs associated with the differential diagnosis of azoospermia.

Xenobiotics with estrogen or antiandrogen action — disruptors of the male reproductive system

Abstract The environmental and life-style changes associated with developing industry and agriculture, especially the exposure to endocrine disrupting chemicals (xenobiotics), are considered as causes of the increasing incidence of male reproductive system disorders. Most of the xenobiotics, which harmfully influence the male reproductive system, reveal estrogen-like (xenoestrogens) or anti-androgenic activity. Recent data have revealed physiological roles of estrogens in the male, however, there are evidences that estrogen-like substances may lead to many undesirable symptoms in the male i.e. gonadal dysgenesis, genital malformations, cryptorchidism, decreased fertility potential and testicular neoplastic changes. The number of xenoestrogens is still growing in the environment, whereas the mechanisms of their action are still not exactly known. They can be harmful not only to the present but potentially also to the next generations.