JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
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2020
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IF 3.209
Mingjie Deng,Feiyan Lin,Caiping Zhou,Yuyan Chen,Leijie Xuan,Hongzhe Wang,Tiantian Feng,Lufeng Hu
Amino acids (AAs) are abundant in seminal fluid and play an important role in the reproduction process. However, the functional significance of AAs in seminal is not clear. This study was designed to profile the AAs in seminal fluid of asthenozoospermia (AS) and oligospermia (OL) patients and evaluate its potential diagnostic value. To determine the true function of AAs in seminal fluid, an ultra high performance liquid chromatography-mass spectrometry (UPLC-MS/MS) was developed and validated. The seminal fluid samples collected from 76 AS patients, 58 OL patients, and 76 healthy subjects were determined by UPLC-MS/MS and analyzed by fisher discriminant and receiver operating characteristic (ROC). The results showed that, the AAs levels were dramatically decreased in AS and OL patients compared with healthy subjects. What’s more there were significant variation in the AAs profile of AS and OL patients. Specifically, some polar AAs decreased, while nonpolar AAs increased in OL patients. Based on the level of AAs, 93.4 % of original and 90.9 % of cross-validated grouped cases were correctly classified between AS patients, OL patients, and healthy subjects. The further ROC analysis shows that glutamic acid, tyrosine, aspartic acid, proline, and tryptophan have the diagnostic value (P < 0.01) in the three groups. Moreover, aspartic acid and glutamic acid exhibit a high correlation (Correlation Coefficient >0.9) in AS, OL, and healthy subjects. In conclusion, there are significant variation in AAs profile in seminal plasma of AS and OL patients with high accuracy in fisher classification. The associations between the levels of AAs in seminal fluid of AS, OL, and healthy subjects may have clinical applications in providing valuable diagnostic indicator.
研究领域:
生殖医学
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代谢组学
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产品关联
DOI
PubMed
高影响因子
HUMAN REPRODUCTION
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2024
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IF 6
Zhang Xiaoning,Liang Min,Song Dandan,Huang Rongzu,Chen Chen,Liu Xiaojun,Chen Houyang,Wang Qingxin,Sun Xiaoli,Song Jian,Zhang Jiali,Kang Hang,Zeng Xuhui
STUDY QUESTIONWhat is the significance and mechanism of human seminal plasma extracellular vesicles (EVs) in regulating human sperm functions?SUMMARY ANSWEREV increases the intracellular Ca2+ concentrations [Ca2+]i via extracellular Ca2+ influx by activating CatSper channels, and subsequently modulate human sperm motility, especially hyperactivated motility, which is attributed to both protein and non-protein components in EV.WHAT IS KNOWN ALREADYEVs are functional regulators of human sperm function, and EV cargoes from normal and asthenozoospermic seminal plasma are different. Pre-fusion of EV with sperm in the acidic and non-physiological sucrose buffer solution could elevate [Ca2+]i in human sperm. CatSper, a principle Ca2+ channel in human sperm, is responsible for the [Ca2+]i regulation when sperm respond to diverse extracellular stimuli. However, the role of CatSper in EV-evoked calcium signaling and its potential physiological significance remain unclear.STUDY DESIGN, SIZE, DURATIONEV isolated from the seminal plasma of normal and asthenozoospermic semen were utilized to investigate the mechanism by which EV regulates calcium signal in human sperm, including the involvement of CatSper and the responsible cargoes in EV. In addition, the clinical application potential of EV and EV protein-derived peptides were also evaluated. This is a laboratory study that went on for more than 5 years and involved more than 200 separate experiments.PARTICIPANTS/MATERIALS, SETTING, METHODSSemen donors were recruited in accordance with the Institutional Ethics Committee on human subjects of the Affiliated Hospital of Nantong University and Jiangxi Maternal and Child Health Hospital. The Flow NanoAnalyzer, western blotting, and transmission electron microscope were used to systematically characterize seminal plasma EV. Sperm [Ca2+]i responses were examined by fluorimetric measurement. The whole-cell patch-clamp technique was performed to record CatSper currents. Sperm motility parameters were assessed by computer-assisted sperm analysis. Sperm hyperactivation was also evaluated by examining their penetration ability in viscous methylcellulose media. Protein and non-protein components in EV were analyzed by liquid chromatography-mass spectrum. The levels of prostaglandins, reactive oxygen species, malonaldehyde, and DNA integrity were detected by commercial kits.MAIN RESULTS AND THE ROLE OF CHANCEEV increased [Ca2+]i via an extracellular Ca2+ influx, which could be suppressed by a CatSper inhibitor. Also, EV potentiated CatSper currents in human sperm. Furthermore, the EV-in [Ca2+]i increase and CatSper currents were absent in a CatSper-deficient sperm, confirming the crucial role of CatSper in EV induced Ca2+ signaling in human sperm. Both proteins and non-protein components of EV contributed to the increase of [Ca2+]i, which were important for the effects of EV on human sperm. Consequently, EV and its cargos promoted sperm hyperactivated motility. In addition, seminal plasma EV protein-derived peptides, such as NAT1-derived peptide (N-P) and THBS-1-derived peptide (T-P), could activate the sperm calcium signal and enhance sperm function. Interestingly, EV derived from asthenozoospermic semen caused a lower increase of [Ca2+]i than that isolated from normal seminal plasma (N-EV), and N-EV significantly improved sperm motility and function in both asthenozoospermic samples and frozen-thawed sperm.LARGE SCALE DATAN/A.LIMITATIONS, REASONS FOR CAUTIONThis was an in vitro study and caution must be taken when extrapolating the physiological relevance to in vivo regulation of sperm.WIDER IMPLICATIONS OF THE FINDINGSOur findings demonstrate that the CatSper-mediated-Ca2+ signaling is involved in EV-modulated sperm function under near physiological conditions, and EV and their derivates are a novel CatSper and sperm function regulators with potential for clinical application. They may be developed to improve sperm motility resulting from low [Ca2+]i response and/or freezing and thawing.STUDY FUNDING/COMPETING INTEREST(S)This research was supported by the National Natural Science Foundation of China (32271167), the Social Development Project of Jiangsu Province (BE2022765), the Nantong Social and People's Livelihood Science and Technology Plan (MS22022087), the Basic Science Research Program of Nantong (JC22022086), and the Jiangsu Innovation and Entrepreneurship Talent Plan (JSSCRC2021543). The authors declare no conflict of interest.
研究领域:
生殖生物学
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细胞生物学
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分子生物学
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生理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Andrology
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2024
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IF 3.2
Yumeng Liu,Qiwen Feng,Liping Zou,Changhong Zhu,Wei Xia
Background Oligoasthenozoospermia is a common cause of male infertility, for which effective treatments are urgently needed. Humanin (HN) is a peptide associated with this condition. Objectives To investigate the ameliorative effect of [Gly14]-Humanin (HNG) on oligoasthenozoospermia and the mechanisms. Materials and methods Mice were treated with cyclophosphamide (CP) to construct a mice model of oligoasthenozoospermia. The resulting model mice were treated with saline or HNG. Subsequently, the testis weights, organ indices, testicular structure, sperm counts and motilities, litter sizes, and serum testosterone concentrations of the mice were determined. Differential gene expression in testicular tissues was determined by RNA sequencing. TM3, TM4, GC1, and GC2 cells were exposed to erastin to induce ferroptosis, followed by treatment with HNG or HNG + ML385 (a nuclear factor erythroid 2-related factor 2 inhibitor). Levels of reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH), and ferrous ions (Fe 2+ ) were determined and their expression of ferroptosis-related proteins was determined by immunofluorescence and western blot. Results The HNG treatment improved testis and sperm parameters and increased litter size and serum testosterone concentrations in model mice. Kyoto Encyclopaedia of Genes and Genomes pathway enrichment analysis revealed significant differential expression of ferroptosis-related genes. The expression of ferroptosis-related proteins increased in testicular tissues after the HNG treatment. The concentrations of ROS, MDA, and Fe 2+ decreased and the concentrations of GSH increased in testicular tissues and in TM3 and TM4 cells after HNG treatment. In vitro experiments confirmed that HNG activated the nuclear factor erythroid 2-related factor 2/glutathione peroxidase 4 (Nrf2/GPX4) pathway. However, these effects of HNG were blocked by ML385 treatment. Discussion and conclusion HNG demonstrated a therapeutic effect on oligoasthenozoospermia in a mouse model by reducing oxidative stress and ferroptosis. In TM3 and TM4 cells, HNG attenuated cellular oxidative stress and inhibited ferroptosis via the Nrf2/GPX4 pathway.
研究领域:
生殖生物学
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分子生物学
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药理学
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF ETHNOPHARMACOLOGY
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2023
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IF 5.4
Mengqi Shi,Tianshi Li,Yan Zhao,Zhongmei He,Ying Zong,Weijia Chen,Rui Du
Ethnopharmacological relevance Antler glue is a classic medicinal to enhance sexual function in traditional Chinese medicine (TCM), which was first recorded in Shen Nong Ben Cao Jing (Shennong′s Classic of the Materia Medica) . Vinegar-processing is a classic method of processing traditional Chinese medicine. The method of preparing antler glue by boiling antlers in vinegar and then concentrating them is recorded in Lei Gong Pao Zhi Lun (Master Lei′s Discourse on Medicinal Processing) . In modern times, the typical processing method of antler glue is water extraction and concentration. However, it is not clear whether there is a difference in the effect of these two processing methods on the chemical composition and pharmacological activity of antler glue. Aim of the study The Chinese Pharmacopoeia (2020) records that the processing method of antler glue is water extraction and concentration. But Lei Gong Pao Zhi Lun differs in Chinese Pharmacopoeia (2020) , which records the processing method of vinegar extraction and concentration. The effect of the two processing methods on antler glue's chemical composition and pharmacological activity is unknown. So this study aimed to elucidate the difference between different processing methods on the chemical composition and the treatment effect on oligoasthenospermia of antler glue. Materials and methods So the automatic amino acid analyzer is used to determine the amino acid content of two different processing methods of antler glue. Proteomics was performed to detect the protein components of two different processing methods of antler glue and analyze them. Cyclophosphamide-induced mice models of oligoasthenospermia were used to study the different pharmacological effects of antler glue in two different processing methods. An automatic sperm analyzer observed the quantity and quality of sperm in mice epididymis. Serum sex hormone testosterone (T), luteinizing hormone (LH) and follicle stimulating hormone (FSH) levels in mice were tested using the enzyme-linked immunosorbent assay (ELISA) kits. Hematoxylin-eosin (H&E) staining was used to analyze pathological alterations in mouse testicular tissue. The transcriptome has been used to reveal the potential mechanism of antler glue in treating oligoasthenospermia. Mitochondrial complex activity assay kits were used to assay the activity of mitochondrial respiratory chain complex I–V in mouse testicular tissue. Western blot was used to determine the expression of related proteins in mouse testicular tissue. Results Vinegar-processing can increase the alanine , proline , and glycine content in antler glue, reduce the length of protein peptides in antler glue, and produce a variety of unique proteins. Vinegar-processed antler glue (VAG) increased sperm density, sperm survival, sperm viability, and serum sex hormone levels in oligozoospermic mice. It reversed testicular damage caused by cyclophosphamide, and the effects were differently superior to those of water-processed antler glue (WAG). In addition, transcriptomics and related experiments have shown that VAG can increase the expression of Ndufa2, Uqcr11, Cox6b1, and Atp5i genes and proteins in mouse testis, thus promoting adenosine diphosphate (ATP) synthesis by increasing the activity of mitochondrial respiratory chain complexes I, III, IV and V. By promoting the oxidative phosphorylation process to produce more ATP, VAG can achieve the therapeutic effect of oligoasthenospermia. Conclusion Vinegar-processing method can increase the content of active ingredients in antler glue. VAG increases ATP levels in the testes by promoting the process of oxidative phosphorylation to treat oligozoospermia.
研究领域:
中医药
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药理学
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生物化学
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生殖医学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）