ABSTRACT
BACKGROUND: The management of male infertility continues to encounter an array of challenges and constraints, necessitating an in-depth exploration of novel therapeutic targets to enhance its efficacy. As an eight-carbon medium-chain fatty acid, octanoic acid (OCA) shows promise for improving health, yet its impact on spermatogenesis remains inadequately researched. METHODS: Mass spectrometry was performed to determine the fatty acid content and screen for a pivotal lipid component in the serum of patients with severe spermatogenesis disorders. The sperm quality was examined, and histopathological analysis and biotin tracer tests were performed to assess spermatogenesis function and the integrity of the blood-testis barrier (BTB) in vivo. Cell-based in vitro experiments were carried out to investigate the effects of OCA administration on Sertoli cell dysfunction. This research aimed to elucidate the mechanism by which OCA may influence the function of Sertoli cells. RESULTS: A pronounced reduction in OCA content was observed in the serum of patients with severe spermatogenesis disorders, indicating that OCA deficiency is related to spermatogenic disorders. The protective effect of OCA on reproduction was tested in a mouse model of spermatogenic disorder induced by busulfan at a dose 30 mg/kg body weight (BW). The mice in the study were separated into distinct groups and administered varying amounts of OCA, specifically at doses of 32, 64, 128, and 256 mg/kg BW. After evaluating sperm parameters, the most effective dose was determined to be 32 mg/kg BW. In vivo experiments showed that treatment with OCA significantly improved sperm quality, testicular histopathology and BTB integrity, which were damaged by busulfan. Moreover, OCA intervention reduced busulfan-induced oxidative stress and autophagy in mouse testes. In vitro, OCA pretreatment (100 µM) significantly ameliorated Sertoli cell dysfunction by alleviating busulfan (800 µM)-induced oxidative stress and autophagy. Moreover, rapamycin (5 µM)-induced autophagy led to Sertoli cell barrier dysfunction, while OCA administration exerted a protective effect by alleviating autophagy. CONCLUSIONS: This study demonstrated that OCA administration suppressed oxidative stress and autophagy to alleviate busulfan-induced BTB damage. These findings provide a deeper understanding of the toxicology of busulfan and a promising avenue for the development of novel OCA-based therapies for male infertility.
Subject(s)
Autophagy , Blood-Testis Barrier , Busulfan , Caprylates , Oxidative Stress , Sertoli Cells , Spermatogenesis , Male , Animals , Blood-Testis Barrier/drug effects , Blood-Testis Barrier/metabolism , Busulfan/adverse effects , Caprylates/pharmacology , Oxidative Stress/drug effects , Mice , Sertoli Cells/drug effects , Sertoli Cells/metabolism , Humans , Spermatogenesis/drug effects , Autophagy/drug effects , Infertility, Male/drug therapy , Infertility, Male/chemically induced , Infertility, Male/pathology , Testis/drug effects , Testis/pathology , Testis/metabolism , Spermatozoa/drug effects , Spermatozoa/metabolism , AdultABSTRACT
BACKGROUND: Polycystic ovary syndrome (PCOS) is a common reproductive endocrine disorder that frequently exhibits low-grade inflammation, pro-oxidant activity, and gut dysbiosis. PCOS has become one of the leading causes of female infertility worldwide. Recently, omega-3 polyunsaturated fatty acids (PUFAs) have been proven to benefit metabolic disorders in PCOS patients. However, its roles in the regulation of metabolic and endocrinal balances in PCOS pathophysiology are not clear. In the present study, we aimed to explore how omega-3 PUFAs alleviate ovarian dysfunction and insulin resistance in mice with dehydroepiandrosterone (DHEA)-induced PCOS by modulating the gut microbiota. METHODS: We induced PCOS in female mice by injecting them with DHEA and then treated them with omega-3 PUFAs. 16S ribosomal DNA (rDNA) amplicon sequencing, fecal microbiota transplantation (FMT) and antibiotic treatment were used to evaluate the role of microbiota in the regulation of ovarian functions and insulin resistance (IR) by omega-3 PUFAs. To further investigate the mechanism of gut microbiota on omega-3-mediated ovarian and metabolic protective effects, inflammatory and oxidative stress markers in ovaries and thermogenic markers in subcutaneous and brown adipose tissues were investigated. RESULTS: We found that oral supplementation with omega-3 PUFAs ameliorates the PCOS phenotype. 16S rDNA analysis revealed that omega-3 PUFA treatment increased the abundance of beneficial bacteria in the gut, thereby alleviating DHEA-induced gut dysbiosis. Antibiotic treatment and FMT experiments further demonstrated that the mechanisms underlying omega-3 benefits likely involve direct effects on the ovary to inhibit inflammatory cytokines such as IL-1ß, TNF-α and IL-18. In addition, the gut microbiota played a key role in the improvement of adipose tissue morphology and function by decreasing multilocular cells and thermogenic markers such as Ucp1, Pgc1a, Cited and Cox8b within the subcutaneous adipose tissues. CONCLUSION: These findings indicate that omega-3 PUFAs ameliorate androgen-induced gut microbiota dysbiosis. The gut microbiota plays a key role in the regulation of omega-3-mediated IR protective effects in polycystic ovary syndrome mice. Moreover, omega-3 PUFA-regulated improvements in the ovarian dysfunction associated with PCOS likely involve direct effects on the ovary to inhibit inflammation. Our findings suggest that omega-3 supplementation may be a promising therapeutic approach for the treatment of PCOS by modulating gut microbiota and alleviating ovarian dysfunction and insulin resistance.
Subject(s)
Dietary Supplements , Fatty Acids, Omega-3 , Gastrointestinal Microbiome , Polycystic Ovary Syndrome , Animals , Female , Mice , Dehydroepiandrosterone/toxicity , Gastrointestinal Microbiome/physiology , Insulin Resistance , Polycystic Ovary Syndrome/chemically induced , Polycystic Ovary Syndrome/drug therapy , Polycystic Ovary Syndrome/physiopathology , Fatty Acids, Omega-3/therapeutic useABSTRACT
In recent years, the postponement of childbearing has become a critical social issue. Male fertility is negatively associated with age because of testis aging. Spermatogenesis is impaired with age, but the molecular mechanism remains unknown. The dynamic posttranslational modification O-linked N-acetylglucosamine (O-GlcNAc), which is a type of monosaccharide modification, has been shown to drive the process of aging in various systems, but it has not yet been investigated in the testis and male reproductive aging. Thus, this study aims to investigate the alteration of O-GlcNAc with aging and explore the role of O-GlcNAc in spermatogenesis. Here, we demonstrate that the decline in spermatogenesis in aged mice is associated with elevation of O-GlcNAc. O-GlcNAc is specifically localized in differentiating spermatogonia and spermatocytes, indicating its crucial role in meiotic initiation and progression. Mimicking the age-related elevation of O-GlcNAc in young mice by disabling O-GlcNAcase (OGA) using the chemical inhibitor Thiamet-G can recapitulate the impairment of spermatogenesis in aged mice. Mechanistically, the elevation of O-GlcNAc in the testis leads to meiotic pachytene arrest due to defects in synapsis and recombination. Furthermore, decreasing O-GlcNAc in aged testes using an O-GlcNAc transferase (OGT) inhibitor can partially rescue the age-related impairment of spermatogenesis. Our results highlight that O-GlcNAc, as a novel posttranslational modification, participates in meiotic progression and drives the impairment of spermatogenesis during aging.
ABSTRACT
OBJECTIVE: To explore the role of postoperative gonadotrophin releasing hormone agonist (GnRH-a) therapy before treatment with intrauterine insemination (IUI) for infertile females with stage I-II endometriosis. MATERIAL AND METHODS: Ninety-seven patients diagnosed with stage I-II endometriosis before IUI were enrolled in this study. The clinical pregnancy rate, cumulative pregnancy rate, live birth rate and newborn conditions were compared between the two groups with and without GnRH-a therapy. RESULTS: The clinical pregnancy rate of IUI in the GnRH-a group was higher than that in the control group (15.29% vs. 11.82%, p = .035). By logistic regression analysis, patients treated with GnRH-a had a higher clinical pregnancy rate than those without (adjusted odds ratio (AOR) 23.190, 95% confidence interval (CI) 1.238-434.312). The live birth rate per IUI cycle in the GnRH-a group was also higher than in the controls (12.94% vs. 10%). However, the difference was not statistically significant (p = .311, AOR 4.844, 95% CI 0.229-102.320). The patients with GnRH-a therapy had a similar incidence of multiple pregnancy rate (0% vs. 0%), miscarriage rate (2.35% vs. 0.91%) and ectopic pregnancy rate (0% vs. 0.91%) as compared to the control group. The cumulative pregnancy rates were all higher in patients administered with GnRH-a than those without GnRH-a treatment in different cycles (one cycle: 17.07% vs 12.50%; two cycles: 29.27% vs 19.64%; three cycles: 31.71% vs 23.21%; ≥four cycles: 31.71% vs 23.21%), but the difference was not statistically significant. Notably, there was no more pregnancy after the third IUI cycle. The gestation weeks of delivery in the two groups were 39.09 ± 1.04 and 38.60 ± 1.17, respectively (p = .323). Nor was there difference in birth weight between the two groups (3236 ± 537 g vs 3435 ± 418 g, p = .360). CONCLUSIONS: The administration of GnRH-a in patients with stage I-II endometriosis could be beneficial to the outcomes of IUI. It is recommended that IUI should be discontinued after three failed attempts. KEY MESSAGESEndometriosis is a common cause of infertility, but the exact mechanism remains unclear.The administration of GnRH-a before IUI treatment is beneficial for patients suffering from stage I-II endometriosis.After three failed attempts, IUI should be stopped in patients with stage I-II endometriosis.
Subject(s)
Endometriosis , Gonadotropin-Releasing Hormone , Endometriosis/drug therapy , Female , Gonadotropin-Releasing Hormone/agonists , Humans , Infant, Newborn , Insemination , Ovulation Induction , Pregnancy , Pregnancy RateABSTRACT
OBJECTIVE: To explore the possible aetiology of subchorionic haematoma (SCH), especially its association with autoantibodies. MATERIAL AND METHODS: Early pregnant women who were detected SCH through ultrasonography were identified as the study group and those without SCH at comparable ages who visited the clinic at the same period of time were compared as the control group. Indexes of laboratory immune tests were compared between the two groups, as well as their pregnancy outcomes. RESULTS: A total of 97 SCH patients and 130 control cases were recruited in this study. A higher proportion of women was detected autoantibodies in the SCH group compared with control group (45.36% vs 21.54%, p = .000). Positive rates of ANA (24.74% vs 10.77%, p = .005) and laboratory antiphospholipid antibodies (ACL, anti-ß2 GP1 or LA) (25.77% vs 11.54%, p = .005) showed significant differences between the two groups. The incidence of vaginal bleeding was significantly higher in the SCH group (43.30% vs 20.00%, p = .000). While the miscarriage rates were not significantly different (17.53% vs 15.38%, p = .666). And there were no significant differences in terms of preterm delivery rate, caesarean section rate, birth weight and pregnancy complications. Most SCHs (96.25%) were absorbed before 20th gestational week. In the SCH group, the average birth weight was significantly lower in women with autoantibodies. Clinical features and other pregnancy outcomes showed no significant differences between SCH patients with and without autoantibodies. CONCLUSIONS: The occurrence of SCH may be associated with autoantibodies. The pregnancy outcomes were comparable between women with and without SCH.KEY MESSAGESSubchorionic haematoma (SCH) is increasingly commonly observed in early pregnancy period, but the aetiology is uncertain and the clinical significance of SCH is controversial.The occurrence of SCH may be associated with autoantibodies.The pregnancy outcomes were not significantly different between women with and without SCH.
Subject(s)
Autoantibodies , Pregnancy Complications , Birth Weight , Cesarean Section , Female , Hematoma/epidemiology , Hematoma/etiology , Humans , Infant, Newborn , Pregnancy , Pregnancy Complications/epidemiology , Pregnancy Outcome/epidemiology , Retrospective StudiesABSTRACT
Ethylene glycol butyl ether (EGBE) is a ubiquitous environmental pollutant that is commonly used in maquillage, industrial, and household products. EGBE has been shown to cause blood toxicity, carcinogenicity, and organ malformations. However, little is known about the impact of EGBE on the female reproductive system, especially oocyte quality. Here, we reported that EGBE influenced oocyte quality by showing the disturbed oocyte meiotic capacity, fertilization potential, and early embryonic development competency. Specifically, EGBE exposure impaired spindle/chromosome structure, microtubule stability, and actin polymerization to result in the oocyte maturation arrest and aneuploidy. In addition, EGBE exposure compromised the dynamics of cortical granules and their component ovastacin, leading to the failure of sperm binding and fertilization. Last, single-cell transcriptome analysis revealed that EGBE-induced oocyte deterioration was caused by mitochondrial dysfunction, which led to the accumulation of ROS and occurrence of apoptosis. Altogether, our study illustrates that mitochondrial dysfunction and redox perturbation is the major cause of the poor quality of oocytes exposed to EGBE.
Subject(s)
Ethylene Glycols/toxicity , Oocytes/drug effects , Animals , Apoptosis/drug effects , Cytoskeleton/drug effects , Cytoskeleton/physiology , DNA Damage , Embryonic Development/drug effects , Environmental Pollutants/toxicity , Female , Meiosis/drug effects , Mice , Organelles/drug effects , Organelles/physiology , Reactive Oxygen SpeciesABSTRACT
BACKGROUND: Spontaneous abortion occurs in 15% ~ 25% of clinical pregnancy. ß-human chorionic gonadotropin (ß-HCG) and progesterone (P) have been widely used in early pregnancy assessment, but their clinical significances are still controversial. Estradiol (E2) has not been used as widely as ß-HCG and P, and its value in predicting pregnancy outcome is unclear. METHODS: In this retrospective study, two hundred early pregnancy women were divided into two groups according to their early pregnancy outcomes: the ongoing pregnancy group and inevitable abortion group. Serum E2 and ß-HCG levels and their growth rates were compared weekly. RESULTS: Estradiol and ß-HCG of the ongoing pregnancy group were significantly higher than that of the inevitable abortion group from the 5th to 10th week of pregnancy. Taking 489.5 pg/mL in the 5th and 6th week, 590.5 pg/mL in the 7th week, and 614.5 pg/mL in the 8th week as cutoff levels of E2, the sensitivity and specificity for E2 to predict bad pregnancy outcome were 91.7% and 41.5%, 82.9% and 71.1%, 84.8% and 84.7%, 75.0% and 95.7%, respectively (P < .05). Both E2 and ß-HCG increased much more rapidly in the ongoing pregnancy group. 80% of the normal pregnancy women showed continuously increasing E2 level. Meanwhile, the inevitable abortion group presented E2 variation types as slow increase or fluctuation, continuous decline, and sudden drop, which account for 54.0%, 34.0%, and 12.0%, respectively. CONCLUSION: Low values and low growth rates of E2 and ß-HCG probably indicate bad pregnancy outcomes.
Subject(s)
Chorionic Gonadotropin, beta Subunit, Human/blood , Estrogens/blood , Pregnancy Trimester, First/blood , Abortion, Spontaneous/epidemiology , Adult , Female , Humans , Pregnancy , Pregnancy Outcome/epidemiology , ROC Curve , Retrospective StudiesABSTRACT
OBJECTIVE: This study aimed to determine the role of uterine artery blood flow (UABF) in recurrent spontaneous abortion (RSA) and to determine a viable option to promote the outcome of pregnancy. METHODS: A total of 421 women were prospectively recruited for this research. UABF indices, including the systolic/diastolic ratio, pulsatility index and resistance index, during the midluteal phase were compared between patients with RSA and women without RSA (controls). After low-molecular-weight heparin (LMWH) was administered to patients with RSA, indices at 7, 10, and 12 weeks of gestation were compared between the two groups. RESULTS: UABF indices during the midluteal phase were significantly higher in the RSA group than in the control group. After LMWH was administered to the RSA group, UABF indices in the first trimester were similar to those in the control group. The rate of spontaneous miscarriage was also similar between the groups after LMWH treatment. CONCLUSIONS: Our study shows that sufficient uterine perfusion is crucial for a successful pregnancy. LMWH reduces the rate of spontaneous miscarriage in patients with RSA to a similar rate in women without RSA. LMWH might play a role in decreasing UABF resistance and increasing uterine perfusion.
Subject(s)
Abortion, Habitual , Abortion, Spontaneous , Abortion, Habitual/drug therapy , Female , Heparin, Low-Molecular-Weight/therapeutic use , Humans , Pregnancy , Prospective Studies , Uterine Artery/diagnostic imaging , Uterus/diagnostic imagingABSTRACT
The aim of this study was to evaluate the effect of virus infection on estradiol (E2) production in human ovarian granulosa cells. Polyriboinosinic polyribocytidylic acid [Poly (I: C)], a synthetic analog of viral double stranded RNA that can be recognized by Toll like receptor 3 (TLR3), was used to imitate virus infection. Granulosa cells (GCs) obtained from patients undergoing in vitro fertilization and embryo transfer (IVF-ET) were cultured in vitro and treated with Poly (I: C), FSH, or both. Concentration of E2 was assayed by electrochemiluminescence. The mRNA and protein expression of TLR3 and aromatase were determined by real-time quantitative PCR (qPCR) and Western blot, respectively. The results showed that expression of TLR3 mRNA was significantly increased after Poly (I: C) stimulation. Poly (I: C) decreased E2 synthesis in FSH-treated GCs. Poly (I: C) inhibited the expression of aromatase in FSH-treated GCs. This study demonstrated that Poly (I: C) inhibits the synthesis of estradiol by granulosa cells under the stimulation of FSH, which might contribute to disturbance of follicular development and ovulation.
Subject(s)
Estradiol/metabolism , Follicle Stimulating Hormone/pharmacology , Granulosa Cells/drug effects , Poly I-C/pharmacology , Adult , Cells, Cultured , Female , Granulosa Cells/metabolism , Granulosa Cells/virology , Humans , Ovarian Follicle/drug effects , Ovarian Follicle/metabolism , Ovarian Follicle/virology , RNA, Viral/pharmacology , Virus Diseases/metabolism , Virus Diseases/pathology , Young AdultABSTRACT
To explore the related factors on the clinical pregnancy outcome in intrauterine insemination, a retrospective study was conducted on the clinical data of 580 cycles for 301 infertile couples who were treated with intrauterine insemination. The female age, male age, duration of infertility, treatment protocols, endometrial thickness and sperm parameters were compared between pregnant group and non-pregnant group. The results showed that there were statistical differences in female age, duration of infertility and endometrial thickness between the two groups. The pregnancy rate was 19.34% in Group A (female age ≤ 30 y) compared with 10.91% in Group B (female age > 30 y). The pregnancy rate was 18.44% when the duration of infertility ≤ 2 years, which was higher than another group 10.73% when the duration of infertility > 2 years. Group analysis according to endometrial thickness (Group1: < 8 mm; Group 2: ≥ 8 mm and ≤ 12 mm; Group 3: > 12 mm) demonstrated significant differences in clinical pregnancy rate (7.41%, 18.00% and 11.48% respectively). For those infertile female without ovulation failure, the higher clinical pregnancy rates were observed in patients undergoing intrauterine insemination in natural cycle 16.12% when compared with the patients in ovarian stimulated cycles 10.48%. Thus, we demonstrate that the pregnancy rate is related with female age, duration of infertility and endometrial thickness. The ovarian stimulated cycle couldn't improve the pregnancy outcome for those women without ovulation disorder in intrauterine insemination.
ABSTRACT
HDAC8 is a class I histone deacetylase that functions in a variety of biological processes through its non-histone substrates. However, its roles during oocyte meiosis remain elusive. Here, we document that HDAC8 localizes at spindle poles and positively participates in the regulation of microtubule organization and spindle assembly in mouse oocytes. Depletion of HDAC8 by siRNA-based gene silencing results in various spindle defects and chromosome misalignment during oocyte meiotic maturation, accompanied by impaired kinetochore-microtubule attachments. Consequently, a higher incidence of aneuploidy is generated in HDAC8-depleted MII eggs. In addition, inhibition of HDAC8 activity with its selective inhibitor PCI-34051 phenocopies the spindle/chromosome defects resulting from HDAC8 depletion by siRNA injection. Finally, we find that HDAC8 is required for the correct localization of Ï-tubulin to spindle poles. Collectively, these data reveal that HDAC8 plays a significant role in regulating spindle assembly and thus ensuring the euploidy in mouse eggs.
Subject(s)
Histone Deacetylases/metabolism , Meiosis/physiology , Oocytes/physiology , Spindle Apparatus/physiology , Aneuploidy , Animals , Cells, Cultured , Chromosome Segregation/drug effects , Female , Histone Deacetylases/chemistry , Hydroxamic Acids/pharmacology , Indoles/pharmacology , Mice , Mice, Inbred ICR , Oocytes/cytology , Oocytes/drug effects , Spindle Apparatus/drug effects , Tubulin/metabolismABSTRACT
OBJECTIVE: To compare the efficacy and safety profile of carbetocin with other uterotonic agents in preventing postpartum hemorrhage. METHODS: PubMed, Web of Science, Scopus and EBSCOhost were searched for relevant randomized controlled trials published until September 2013. RESULTS: Carbetocin was associated with a significantly reduced need for additional uterotonic agents (RR = 0.68, 95% CI: 0.55-0.84, I(2 )= 4%) compared with oxytocin in women following cesarean delivery. However, with respect to postpartum hemorrhage, severe postpartum hemorrhage, mean estimated blood loss and adverse effects, our analysis failed to detect a significant difference. Studies comparing carbetocin with syntometrine in women undergoing vaginal delivery demonstrated no statistical difference in terms of risk of postpartum hemorrhage, severe postpartum hemorrhage or the need for additional uterotonic agents, but the risk of adverse effect was significantly lower in the carbetocin group. CONCLUSIONS: Carbetocin has been associated with a similar low incidence of adverse effects to oxytocin and at least as effective as syntometrine and may become an alternative uterotonic agent for the prevention of postpartum hemorrhage. Further studies should be conducted to determine the safety and efficacy profile of carbetocin in women with cardiac disorders and to analyze the cost-effectiveness and minimum effective dose of carbetocin.
Subject(s)
Oxytocics/therapeutic use , Oxytocin/analogs & derivatives , Postpartum Hemorrhage/drug therapy , Female , Humans , Oxytocin/therapeutic use , Pregnancy , Randomized Controlled Trials as TopicABSTRACT
The mature oocyte contains the full complement of maternal proteins required for fertilization, the transition to zygotic transcription, and the beginning stages of embryogenesis. Many of these proteins have yet to be characterized. In this study, two-dimensional electrophoresis (2-DE) of mouse metaphase-II (MII) oocyte proteins, stained with silver staining or Pro-Q Diamond dye, was performed to describe the proteome and phosphoproteome of the mouse oocyte derived from ICR mice. A total of 869 selected protein spots, corresponding to 380 unique proteins, were identified successfully by mass spectrometry, in which 90 protein spots representing 53 unique proteins have been stained with Pro-Q Diamond, indicating that they are in phosphorylated forms. All identified proteins were bioinformatically annotated in detail and compared with the embryonic stem cell (ESC) proteome. A proteome reference database for the mouse oocyte was established from the protein data generated in this study, which can be accessed over the Internet ( http://reprod.njmu.edu.cn/2d). This database is the most detailed mouse oocyte proteomic database to date. It should be valuable in expanding our knowledge of the regulation of signaling in oogenesis, fertilization, and embryo development, while revealing potential mechanisms for epigenetic reprogramming.
Subject(s)
Meiosis , Metaphase , Oocytes/metabolism , Phosphoproteins/metabolism , Proteome/analysis , Animals , Computational Biology/methods , Databases, Protein , Female , Mice , Mice, Inbred ICR , Oocytes/chemistry , Proteome/metabolism , Proteomics/methods , Silver StainingABSTRACT
AIM: To investigate the expression of Spindlin 1 (Spin 1) isoform2 and assess its function in mouse testis. METHODS: First, reverse-transcription polymerase chain reaction (RT-PCR) was used to determine whether Spin1 isoform2 is present in mouse testis. Then the expression patterns of the isoform between newborn and adult mice testes were compared by immunoblot analysis. Finally, the diversity of its localization in mice testes at different ages (days 0, 7, 14, 21, 28 and 60) was observed by immunohistochemistry. The localization of the protein in mouse sperm was also investigated by immunofluorescence. RESULTS: The RT-PCR results show that Spin1 isoform2 is present in mouse testis. As shown by immunoblot analysis, the isoform was more highly expressed in adult testes compared with newborn testes. Interestingly, Spin1 isoform2 did not show up in the cytoplasm of primary spermatocytes until day 14. Also, the protein exists at the tail of the mouse sperm. CONCLUSION: Spin1 isoform2 is a protein expressed highly in adult testis, which might be involved in spermatogenesis and could be necessary for normal sperm motility.
Subject(s)
Cell Cycle Proteins/genetics , Cell Cycle Proteins/metabolism , Microtubule-Associated Proteins/genetics , Microtubule-Associated Proteins/metabolism , Phosphoproteins/genetics , Phosphoproteins/metabolism , Testis/physiology , Age Factors , Animals , Animals, Newborn , Blotting, Western , Cytoplasm/metabolism , Female , Immunohistochemistry , Male , Meiosis/physiology , Mice , Mice, Inbred ICR , Pregnancy , Reverse Transcriptase Polymerase Chain Reaction , Sperm Motility/physiology , Spermatogenesis/physiologyABSTRACT
Hemin-catalytic decomposition of artemisinin (qinghaosu, QHS) was studied using pyronine B (PB) as an indicator. The interaction between hemin and QHS was an enzyme-substrate model, and the action sites were the endoperoxide moiety of QHS and the central metal ion of enzyme respectively. The kinetic catalytic constant depends upon enzyme and substrate concentrations, and the Michaelis-Menten parameters Km, Vmax and Kcat was 8.4 x 10(-5) mol x L(-1), 7.4 x 10(-6) mol x L(-1) s(-1) and 50.23 s(-1) respectively. The catalytic activity of hemin was inhibited in the presence of deactivated agents and at high temperature. Under optimal conditions, the change in fluorescence intensity (Fo-F) of pyronine B was proportional to the QHS concentration from 0.0 to 1.27 x 10(-6) mol x L(-1), and the detection limit (3sigma) was as low as 2.3 x 10(-8) mol x L(-1). The proposed method was applied to detect the concentration of QHS in the media of plasma and urine.