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1.
Open Vet J ; 11(1): 70-79, 2021.
Article in English | MEDLINE | ID: mdl-33898286

ABSTRACT

Background: Male infertility has been on the rise since the past seven decades. Recently, in Libya, bee venom therapy (BVT) has become a popular method among alternative healthcare practitioners for treating male infertility. However, a literature search did not find any published studies that investigated the use of BVT for infertility treatment. Aim: To investigate the effect of bee venom on the male reproductive status through measurements of semen quality parameters and testicular histological changes in adult male mice. Methods: A total of 48 male mice were randomly divided into three experimental groups (which were subdivided into two subgroups with eight mice each) as follows: control, bee venom sting (BVS), and bee venom injection (BVI). The normal control subgroup mice were not subjected to any treatment, while the vehicle control subgroup mice were injected (i.p.) with 200 µl of 0.9% saline solution. In the BVS-treated subgroups, each mouse was stung by one live bee for five times (BVS-5) or seven times (BVS-7) every third day for 2 or 3 weeks. While each mouse in the BVI-treated subgroups received 23 µg/kg in a dose volume of 200 µl BVIs (i.p.) for five times (BVI-5) or seven times (BVI-7) every third day for 15 or 21 days. Results: The findings of this study showed that repeated bee venom treatment by sting or injection to adult male mice resulted in a significant decline in testosterone levels, sperm count, sperm motility, and a very significant increase in the percentage of abnormal sperm morphology; also, there were harmful testicular histological changes in the structural organization of seminiferous tubules and degenerative changes in the germinal epithelium compared to control group. Conclusion: The results of this study provide evidence for the low semen quality and adverse testicular histological changes in male mice treated with bee venom. Hence, there is a desperate need for educating alternative healthcare practitioners and infertile couples about the harmful effects of BVT on reproductive status.


Subject(s)
Bee Venoms/administration & dosage , Fertility Agents, Male/administration & dosage , Mice/physiology , Semen Analysis , Testis/drug effects , Animals , Bee Venoms/adverse effects , Bee Venoms/pharmacology , Fertility Agents, Male/adverse effects , Fertility Agents, Male/pharmacology , Injections, Intraperitoneal/statistics & numerical data , Insect Bites and Stings/complications , Male , Random Allocation , Testis/anatomy & histology , Testis/physiology
2.
Methods Mol Biol ; 2240: 65-76, 2021.
Article in English | MEDLINE | ID: mdl-33423227

ABSTRACT

Contraction of cauda epididymal duct (CE) smooth muscle is one of the very first events of the seminal emission phase of ejaculation. The contraction of CE smooth muscle is governed by a complex interaction of hormones, autacoids, and by the neurotransmitters released from the epididymal intramural nerve endings, and any impairment in the CE smooth muscle contraction has the potential to impair male fertility. Apart the obvious pathophysiological and toxicological importance of CE smooth muscle contraction, modulation of CE contraction has pharmaceutical interest offering a druggable target to development of drugs to improve/impair male fertility. The in vitro contraction experiments constitute a valuable approach to an in-depth evaluation of functional and molecular changes resulting from pathologies or drug exposure. Therefore, this chapter consists in a description of in vitro pharmacological reactivity contractility of the epididymal duct in a controlled medium, maintained at 30 °C of temperature and continuously bubbled with 95% O2 and 5% CO2 to obtain cumulative concentration-response curves that has been fundamental to some of our investigations on epididymal physiology, toxicology, and pharmacology.


Subject(s)
Drug Evaluation, Preclinical/methods , Epididymis/drug effects , Fertility Agents, Male/pharmacology , Muscle Contraction , Animals , Drug Evaluation, Preclinical/instrumentation , Epididymis/physiology , Male , Muscle, Smooth/physiology , Rats
3.
Biomed Pharmacother ; 133: 111085, 2021 Jan.
Article in English | MEDLINE | ID: mdl-33378981

ABSTRACT

Obesity is a significant global health and socio-economic challenge, and considered an important risk factor for poor health outcomes including male reproductive dysfunction and infertility. As excess adiposity causes testicular dysfunction and infertility, novel therapeutic strategies require investigation. Nigella sativa (Ns) seed oil and metformin have both demonstrated a potential positive effect on obesity, although both remain poorly investigated in male fertility. Therefore, this study aimed to determine the effect of Ns oil and metformin on total body weight (TBW), mitochondrial membrane potential (MMP), serum testosterone and semen parameters in an obese animal model. Wistar rats (n = 54) were divided into six groups: normal chow (NC), high sugar diet (HSD) only, HSD and saline, HSD and metformin (75 mg/Kg/day), HSD and Ns (200 mg/Kg/day) (NS200), HSD and Ns (400 mg/Kg/day) (NS400). Intervention was force fed for the last 8 weeks of the 14 weeks dietary exposures. Results showed that the HSD increased TBW (P = 0.001) and reduced sperm concentration (P = 0.013) and progressive motility (P = 0.009) compared to the NC group. Metformin, NS200 and NS400 improved TBW (P = 0.035, P = 0.006 and P = 0.005, respectively) and testosterone (P < 0.001) compared to the HSD saline group, where metformin and NS400 improved sperm concentration (P < 0.001 and P = 0.049, respectively) and MMP (P < 0.001). There were no changes in sperm motility and viability for all experimental exposures, although NS400 (P = 0.047) negatively affected sperm viability. Metformin and Ns may be novel treatment options in obesity-induced infertility, although a potential negative impact on viability is cautioned for high dose Ns. These results warrant further investigation of Ns and Metformin for the management of obese infertile males.


Subject(s)
Anti-Obesity Agents/pharmacology , Fertility Agents, Male/pharmacology , Fertility/drug effects , Infertility, Male/prevention & control , Metformin/pharmacology , Obesity/drug therapy , Plant Oils/pharmacology , Spermatozoa/drug effects , Testosterone/blood , Animals , Biomarkers/blood , Disease Models, Animal , Infertility, Male/blood , Infertility, Male/pathology , Infertility, Male/physiopathology , Male , Membrane Potential, Mitochondrial/drug effects , Obesity/blood , Obesity/pathology , Rats, Wistar , Sperm Count , Sperm Motility/drug effects , Spermatozoa/metabolism , Spermatozoa/pathology
4.
Reprod Sci ; 28(2): 371-380, 2021 02.
Article in English | MEDLINE | ID: mdl-32780358

ABSTRACT

Spermatogenesis process is sensitive to heat stress because the testicular temperature is 2 to 4 °C lower than the core body temperature. The current study aimed to investigate the effects of iron oxide nanoparticles containing curcumin on spermatogenesis in mice induced by long-term scrotal hyperthermia. In this experimental study, 18 mice were equally divided into the following three groups: control, scrotal hyperthermia, and scrotal hyperthermia + curcumin-loaded iron particles (NPs) (240 µL) (mice were treated for 20 days). Hyperthermia was induced by exposure to the temperature of 43 °C for 20 min every other day for 5 weeks. Afterward, the animals were euthanized; sperm samples were collected for sperm parameters analysis, and testis samples were taken for histopathology experiments, evaluation of serum testosterone level, and RNA extraction in order to examine the expression of c-kit, STRA8 and PCNA genes. Our study showed that curcumin-loaded iron particles could notably increase the volume of testis, length of seminiferous tubules, sperm parameters, and stereological parameters (i.e., spermatogonia, primary spermatocyte, round spermatid, and Leydig cells) thereby increasing serum testosterone level; in addition, TUNEL-positive cells showed a significant decrease in curcumin-loaded iron particle group. Thus, based on the obtained results, the expression of c-kit, STRA8, and PCNA genes was significantly increased in treatment groups by curcumin-loaded iron particles compared with scrotal hyperthermia-induced mice. In conclusion, curcumin-loaded iron particles can be considered an alternative treatment for improving the spermatogenesis process in scrotal hyperthermia-induced mice.


Subject(s)
Azoospermia/drug therapy , Curcumin/pharmacology , Drug Carriers , Fertility Agents, Male/pharmacology , Magnetic Iron Oxide Nanoparticles/chemistry , Spermatogenesis/drug effects , Spermatozoa/drug effects , Testis/drug effects , Adaptor Proteins, Signal Transducing/genetics , Adaptor Proteins, Signal Transducing/metabolism , Animals , Azoospermia/blood , Azoospermia/etiology , Azoospermia/pathology , Biomarkers/blood , Curcumin/chemistry , Disease Models, Animal , Drug Compounding , Fertility Agents, Male/chemistry , Hyperthermia, Induced , Male , Mice , Proliferating Cell Nuclear Antigen/genetics , Proliferating Cell Nuclear Antigen/metabolism , Proto-Oncogene Proteins c-kit/genetics , Proto-Oncogene Proteins c-kit/metabolism , Spermatozoa/metabolism , Spermatozoa/pathology , Testis/metabolism , Testis/pathology , Testis/physiopathology , Testosterone/blood , Time Factors
5.
Biomed Pharmacother ; 133: 110868, 2021 Jan.
Article in English | MEDLINE | ID: mdl-33181455

ABSTRACT

Male infertility has become an important health problem that is primarily caused by testicular dysfunction with abnormal spermatogenesis. In this study, we demonstrated that the neuropeptide, substance P (SP), is essential for spermatogonia proliferation in a seminiferous tubule culture system. In addition, SP (5 nmol/kg) treatment markedly restored spermatogenesis, improved sperm quality, and increased the number of ZBTB16+ or LIN28+ undifferentiated spermatogonia as well as STRA8+ differentiated spermatogonia in a busulfan-induced non-obstructive azoospermic mouse model. Furthermore, 100 nM SP treatment in vitro significantly stimulated the proliferation of GC-1 spg cells (a spermatogonia cell line) via activation of the Erk1/2 signaling pathway. Moreover, the sperm quality and the number of spermatogonia were significantly reduced after treatment with RP67580, a selective NK-1 receptor antagonist, suggesting that SP-NK1R signaling plays an important role in spermatogenesis. Taken together, these results suggest that SP may be a potential therapeutic agent for male infertility by accelerating the restoration of spermatogenesis.


Subject(s)
Azoospermia/drug therapy , Fertility Agents, Male/pharmacology , Fertility/drug effects , Spermatogenesis/drug effects , Spermatogonia/drug effects , Substance P/pharmacology , Animals , Apoptosis/drug effects , Azoospermia/chemically induced , Azoospermia/metabolism , Azoospermia/physiopathology , Busulfan , Cell Line , Cell Proliferation/drug effects , Disease Models, Animal , Extracellular Signal-Regulated MAP Kinases/metabolism , Male , Mice, Inbred C57BL , Receptors, Neurokinin-1/agonists , Receptors, Neurokinin-1/metabolism , Signal Transduction , Spermatogonia/metabolism , Tissue Culture Techniques
6.
BMC Complement Altern Med ; 19(1): 132, 2019 Jun 17.
Article in English | MEDLINE | ID: mdl-31208410

ABSTRACT

BACKGROUND: Nigerian Cannabis sativa (hemp) causes male gonadotoxicity by inducing hyperprolactinemia, down-regulation of hypothalamic-pituitary-testicular axis, and oxidative stress. Benin republic hemp has been preferred by illicit users in Nigeria but its effect on male fertility is not understood. This study determined and compared the compositions of Benin republic hemp ethanol extract (BHE) and Nigerian hemp. The effects of BHE on semen parameters, reproductive hormones, and anti-oxidant status, and the possibility of bromocriptine (prolactin inhibitor) to abolish hemp-induced toxicities in rats were also investigated. METHODS: Thirty-six male Wistar rats were blindly randomized into 6 oral treatment groups (n = 6 each). Groups I (control) and II received normal saline and bromocriptine (3 mg/kg) respectively. Groups III and IV received 2 mg/kg of BHE alone and in combination with bromocriptine respectively, while groups V and VI received 10 mg/kg BHE alone and in combination with bromocriptine respectively. Comparisons among the groups were done by one-way analysis of variance, followed by post-hoc Tukey multiple comparison test. Statistical significance was considered at p < 0.05. RESULTS: The BHE has no cannabichromene and tetrahydrocannabinol but a very small quantity of cannabinol and higher quantity of fatty acids when compared to Nigerian hemp. Both doses of BHE increased sperm count, morphology and viability but not motility. Co-administration of BHE with bromocriptine lowered sperm count but increased sperm morphology and viability. Bromocriptine and/or BHE caused reduction in the plasma prolactin level, increase in the plasma superoxide dismutase activity, but no significant change in the plasma gonadotropin releasing hormone, follicle stimulating hormone (except for the increase in rats that received bromocriptine+ 10 mg/kg BHE), luteinizing hormone, estradiol, malondialdehyde and glutathione peroxidase. The 10 mg/kg BHE or bromocriptine+BHE (both doses) increased total anti-oxidant capacity and catalase. CONCLUSIONS: The BHE improves semen parameters by reducing plasma prolactin and enhancing plasma anti-oxidant status. Its pro-fertility potential might be associated with its deficiency in the widely known gonadotoxic phytocannabinoids.


Subject(s)
Antioxidants/metabolism , Cannabinoids/analysis , Cannabis/chemistry , Dronabinol/analysis , Fertility Agents, Male/pharmacology , Plant Extracts/pharmacology , Prolactin/metabolism , Semen/drug effects , Animals , Cannabinoids/pharmacology , Dronabinol/pharmacology , Fertility/drug effects , Fertility Agents, Male/analysis , Male , Plant Extracts/analysis , Rats , Semen/metabolism
7.
Reprod Domest Anim ; 54(2): 184-194, 2019 Feb.
Article in English | MEDLINE | ID: mdl-30179285

ABSTRACT

Conventional in vitro fertilization has not yet been implemented in the equine species. One of the main reasons has been the inability to develop a culture medium and incubation conditions supporting high levels of stallion sperm capacitation and hyperactivation in vitro. Although different culture media have been used for this purpose, human tubal fluid (HTF) medium, widely used in the manipulation of human and mice gametes, has not been reported so far in stallion sperm culture. The first part of this study aimed to compare HTF and Whitten's media on different stallion sperm quality and capacitation variables. Additionally, the effect of procaine, aminopyridine and caffeine in both media was evaluated on sperm motility parameters at different incubation times. Integrity and destabilization of the plasma membrane were evaluated by merocyanine 540/SYTOX Green (MC540), mitochondrial membrane potential (∆Ψm) using tetramethylrhodamine methyl ester perchlorate (TMRM), acrosome membrane integrity by PNA/FITC and tyrosine phosphorylation by P-tyrosine mouse mAb conjugated to Alexa Fluor® by flow cytometry. Motility parameters were evaluated using the integrated semen analysis system (ISAS®). We found no differences between Whitten's and HTF media and incubation time in terms of sperm viability, uninduced acrosome membrane damage or mitochondrial membrane potential at 30- and 120-min incubation. Membrane fluidity (MC540) increased in both media at 30- and 120-min incubation compared to noncapacitating conditions. Similarly, tyrosine phosphorylation increased in both media in capacitating conditions at 2- and 4-hr incubation compared to noncapacitating conditions. Although procaine showed the best result in terms of sperm hyperactivated motility in both media, aminopyridine also showed parameters consistent with the hyperactivation including an increase in curvilinear velocity and decrease in straightness. In conclusion, HTF medium and aminopyridine equally support capacitation-related parameters in stallion sperm.


Subject(s)
Culture Media/pharmacology , Fertility Agents, Male/pharmacology , Horses , Semen Analysis/veterinary , Sperm Capacitation/drug effects , Sperm Motility/drug effects , Acrosome Reaction/drug effects , Aminopyridines/pharmacology , Animals , Caffeine/pharmacology , Fertilization in Vitro/veterinary , Fluoresceins/pharmacology , Male , Membrane Potential, Mitochondrial , Peanut Agglutinin/pharmacology , Phosphorylation , Procaine/pharmacology , Semen/drug effects , Semen Analysis/methods , Spermatozoa/drug effects
8.
PLoS One ; 13(6): e0197897, 2018.
Article in English | MEDLINE | ID: mdl-29856778

ABSTRACT

Membrane integrity is essential in maintaining sperm viability, signaling, and motility, which are essential for fertilization. Sperm are highly susceptible to oxidative stress, as they are rich in sensitive polyunsaturated fatty acids (PUFA), and are unable to synthesize and repair many essential membrane constituents. Because of this, sperm cellular membranes are important targets of this process. Membrane Lipid Replacement (MLR) with glycerophospholipid mixtures (GPL) has been shown to ameliorate oxidative stress in cells, restore their cellular membranes, and prevent loss of function. Therefore, we tested the effects of MLR on sperm by tracking and monitoring GPL incorporation into their membrane systems and studying their effects on sperm motility and viability under different experimental conditions. Incubation of sperm with mixtures of exogenous, unoxidized GPL results in their incorporation into sperm membranes, as shown by the use of fluorescent dyes attached to GPL. The percent overall (total) sperm motility was increased from 52±2.5% to 68±1.34% after adding GPL to the incubation media, and overall sperm motility was recovered from 7±2% after H2O2 treatment to 58±2.5%)(n = 8, p<0.01) by the incorporation of GPL into sperm membranes. When sperm were exposed to H2O2, the mitochondrial inner membrane potential (MIMP), monitored using the MIMP tracker dye JC-1 in flow cytometry, diminished, whereas the addition of GPL prevented the decrease in MIMP. Confocal microscopy with Rhodamine-123 and JC-1 confirmed the mitochondrial localization of the dyes. We conclude that incubation of human sperm with glycerolphospholipids into the membranes of sperm improves sperm viability, motility, and resistance to oxidizing agents like H2O2. This suggests that human sperm might be useful to test innovative new treatments like MLR, since such treatments could improve fertility when it is adversely affected by increased oxidative stress.


Subject(s)
Glycerophospholipids/chemistry , Glycerophospholipids/pharmacology , Micelles , Oxidative Stress/drug effects , Sperm Motility/drug effects , Spermatozoa/cytology , Spermatozoa/drug effects , Adult , Cell Membrane/drug effects , Cell Membrane/metabolism , Culture Techniques , Fertility Agents, Male/chemistry , Fertility Agents, Male/pharmacology , Glycerophospholipids/metabolism , Humans , Male , Spermatozoa/metabolism
9.
Reprod Biol ; 15(2): 126-9, 2015 Jun.
Article in English | MEDLINE | ID: mdl-26051462

ABSTRACT

The purpose of the current study was to investigate the effects of N-acetyl-L-cysteine (NAC) and catalase (CAT) on chicken sperm parameters during liquid storage for up to 48 h at 5 °C. Supplementation of EK extender with NAC (15 mM) increased sperm motility after 24h. After 48 h, an increase in sperm viability with NAC (5, 15 mM) and CAT (100, 300 U/mL) was observed, but only treatment with 15 mM NAC improved sperm progressive motility.


Subject(s)
Acetylcysteine/pharmacology , Antioxidants/pharmacology , Fertility Agents, Male/pharmacology , Semen Preservation/veterinary , Spermatozoa/drug effects , Animals , Animals, Inbred Strains , Catalase/metabolism , Cell Survival/drug effects , Chickens , Male , Osmolar Concentration , Poland , Refrigeration/veterinary , Sperm Motility/drug effects , Spermatozoa/cytology , Time Factors
10.
Int J Biol Macromol ; 80: 16-22, 2015 Sep.
Article in English | MEDLINE | ID: mdl-26093318

ABSTRACT

In this study, purification, preliminary characterization and biological activities of water-soluble polysaccharides from Ostrea rivularis Gould (ORP) were investigated. Firstly, crude ORP was extracted by enzyme-assisted extraction and then sequentially purified by chromatography of DEAE-52 and Sephadex G-100, producing one main purified fractions of ORPp. Furthermore, the preliminary characterization of ORPp was studied, and its antioxidant and spermatogenesis activities were evaluated. Experimental results showed that ORPp was mainly composed of glucose (76.3%) and galactose (23.7%). The average molecular weight of ORPp was 118 kDa. Besides, ORPp showed strong antioxidant activities in vitro. For the experiments of antioxidant activities in vivo, ORPp can significantly inhibited the formation of MDA in rats' serums, and raised the activities of antioxidant enzymes and the level of total antioxidant capacity (TAOC). Furthermore, ORPp could significantly increase the weights of male rats' sexual organs, promote sperm motility and raise epididymal sperm counts. These results suggest that ORPp could be a new source of natural antioxidants and spermatogenic agent with its potential usage in developing novel supplements and medicines.


Subject(s)
Fertility Agents, Male/pharmacology , Free Radical Scavengers/pharmacology , Ostrea/chemistry , Polysaccharides/pharmacology , Animals , Drug Evaluation, Preclinical , Fertility Agents, Male/chemistry , Fertility Agents, Male/isolation & purification , Free Radical Scavengers/chemistry , Free Radical Scavengers/isolation & purification , Free Radicals/chemistry , Male , Mice , Organ Size/drug effects , Oxidation-Reduction , Polysaccharides/chemistry , Polysaccharides/isolation & purification , Rats, Wistar , Sperm Count , Sperm Motility , Spermatogenesis/drug effects , Superoxides/chemistry
11.
Reprod Domest Anim ; 49(6): 894-8, 2014 Dec.
Article in English | MEDLINE | ID: mdl-25132093

ABSTRACT

Poor reproductive performance of Merino ewe flocks when mated to Border Leicester rams during spring may be due to seasonality of the Border Leicester breed. Two approaches were taken to test this assumption. Six young (12 months old) or six mixed-age (12, 24 and ≥36 months old) Border Leicester rams were either treated or not treated with melatonin implants (2 × 2 design) 6 weeks before the four groups of rams were each put with approximately 300 Merino ewes for an 8-week mating period. Implants were inserted in early September (experiment 1). The second approach was to yard or not yard ewes and mixed-age rams on several occasions during the first 3 weeks of the mating period (experiment 2). Pregnancy rate and twinning percentage were assessed by ultrasonography. In experiment 1, melatonin treatment in young rams increased (p < 0.001) pregnancy rate from 5.0% to 92.6%, but mixed-age rams did not respond (90.7% vs 89.5% for melatonin and non-melatonin treatments, respectively). Twinning rate was similar (p > 0.05) for ewes mated to either melatonin or non-melatonin-treated young rams (36.8% vs 40.0%, respectively), whereas melatonin significantly improved (p < 0.05) twinning rate in those ewes mated to mixed-age rams (49.1% vs 36.1%). After 6 weeks of melatonin treatment, scrotal circumference was greater (p < 0.05) in both young and mixed-aged rams than in untreated counterparts. In experiment 2, yarding of ewes and rams overnight on several occasions early in the mating period reduced (p < 0.001) pregnancy rate compared with non-yarded counterparts (89.5% vs 65.5%). Twinning rate was not affected (37.7% vs 36.1%, respectively). In summary, melatonin treatment of Border Leicester rams significantly improved flock reproductive performance in spring due to improved pregnancy rates with young rams and improved litter size with mixed-age rams.


Subject(s)
Fertility Agents, Male/pharmacology , Melatonin/pharmacology , Sheep/physiology , Animals , Drug Implants , Female , Male , Pregnancy , Pregnancy Rate , Pregnancy, Multiple/drug effects , Seasons
12.
Molecules ; 19(7): 9502-14, 2014 Jul 03.
Article in English | MEDLINE | ID: mdl-24995929

ABSTRACT

The present study investigated the effects and potential mechanism(s) of action of icariin on the reproductive functions of male rats. Adult rats were treated orally with icariin at doses of 0 (control), 50, 100, or 200 mg/kg body weight for 35 consecutive days. The results show that icariin had virtually no effect on the body weight or organ coefficients of the testes or epididymides. However, 100 mg/kg icariin significantly increased epididymal sperm counts. In addition, 50 and 100 mg/kg icariin significantly increased testosterone levels. Real-time PCR suggests icariin may be involved in testosterone production via mRNA expression regulation of genes such as peripheral type benzodiazepine receptor (PBR) and steroidogenic acute regulatory protein (StAR). Furthermore, 100 mg/kg icariin treatment also affected follicle stimulating hormone receptor (FSHR) and claudin-11 mRNA expression in Sertoli cells. Superoxide dismutase (SOD) activity and malondialdehyde (MDA) levels were measured in the testes; 50 and 100 mg/kg icariin treatment improved antioxidative capacity, while 200 mg/kg icariin treatment upregulated oxidative stress. These results collectively suggest that icariin within a certain dose range is beneficial to male reproductive functions; meanwhile, higher doses of icariin may damage reproductive functions by increasing oxidative stress in the testes.


Subject(s)
Fertility Agents, Male/pharmacology , Flavonoids/pharmacology , Spermatogenesis/drug effects , Animals , Drug Evaluation, Preclinical , Gene Expression/drug effects , Male , Malondialdehyde/metabolism , Organ Specificity , Rats, Sprague-Dawley , Receptors, LH/genetics , Receptors, LH/metabolism , Sperm Count , Superoxide Dismutase/metabolism , Testis/cytology , Testis/drug effects , Testis/physiology , Testosterone/blood
13.
Endocrinology ; 154(10): 3914-24, 2013 Oct.
Article in English | MEDLINE | ID: mdl-23885018

ABSTRACT

The molecular mechanism of the aging-associated dysfunction of Leydig cells (LCs) is complex and poorly understood. In this study, we analyzed the contribution of nitric oxide (NO) and cGMP signaling to the age-dependent decline in LC function. Significant (>50%) decreases in serum, intratesticular, and LC androgens in aging rats (15-24 months) were accompanied by a proportional increase in NO production, an up-regulation of cGMP levels, and the expression of soluble guanylyl cyclase-1B and protein kinase G1 in LCs. In contrast, LC cAMP levels decreased with age, most likely reflecting the up-regulation of cAMP-specific phosphodiesterase expression. Moreover, the expression of genes encoding enzymes responsible for cholesterol transport and its conversion to T were reduced. Exposing LCs from aged animals to NO further increased cGMP levels and decreased cAMP and androgen production, whereas the addition of cell-permeable 8-bromoguanosine-cGMP alone had the opposite effect. In vivo inhibition of cGMP-specific phosphodiesterase-5 for 3 and 6 months in aged rats led to a partial restoration of androgens, NO, and cyclic nucleotide levels, as well as the expression of steroidogenic and NO/cGMP signaling genes. These results indicate that a progressive increase in NO production contributes to the age-dependent decrease in steroidogenesis in a cGMP-independent manner, whereas the sustained elevation in cGMP levels significantly slows the decline in LC function.


Subject(s)
Aging , Androgens/metabolism , Cyclic AMP/metabolism , Leydig Cells/metabolism , Nitric Oxide/metabolism , Second Messenger Systems , Testis/metabolism , Androgens/blood , Animals , Cells, Cultured , Cyclic GMP-Dependent Protein Kinase Type I/biosynthesis , Cyclic GMP-Dependent Protein Kinase Type I/genetics , Cyclic GMP-Dependent Protein Kinase Type I/metabolism , Cyclic Nucleotide Phosphodiesterases, Type 5/chemistry , Cyclic Nucleotide Phosphodiesterases, Type 5/metabolism , Fertility Agents, Male/pharmacology , Gene Expression Regulation, Developmental/drug effects , Guanylate Cyclase/biosynthesis , Guanylate Cyclase/genetics , Guanylate Cyclase/metabolism , Leydig Cells/cytology , Leydig Cells/drug effects , Male , Phosphodiesterase 5 Inhibitors/pharmacology , Rats , Rats, Wistar , Receptors, Cytoplasmic and Nuclear/biosynthesis , Receptors, Cytoplasmic and Nuclear/genetics , Receptors, Cytoplasmic and Nuclear/metabolism , Second Messenger Systems/drug effects , Soluble Guanylyl Cyclase , Testis/cytology , Testis/drug effects , Testis/growth & development , Up-Regulation
14.
Contraception ; 84(1): 98-107, 2011 Jul.
Article in English | MEDLINE | ID: mdl-21664518

ABSTRACT

BACKGROUND: The aim of the present study was to evaluate the contraceptive efficacy of Cuminum cyminum (jeera) seeds in male albino rats. STUDY DESIGN: C. cyminum methanol extract (CcMtE) at dose levels of 100 and 200 mg/rat/day was orally administered to male rats for 60 days. The effect of the treatment on reproductive organs and fertility was investigated. Recovery and toxicity studies were also carried out. RESULTS: C. cyminum methanol extract fed to male rats for 60 days did not cause any alterations in the body weight, whereas the weight of testes, epididymides, seminal vesicles and ventral prostate were significantly reduced (p≤.001). Animals treated with CcMtE showed a marked reduction in sperm density in the cauda epididymis and testes and sperm motility in the cauda epididymis. Reduction in fertility was 69.0% and 76.0% in 100 and 200 mg/rat/day dose levels, respectively. The circulatory hormones were also reduced significantly. Testicular biochemical analysis of protein, sialic acid, glycogen, ascorbic acid and fructose indicated a marked decline, whereas testicular cholesterol content was significantly increased, which showed altered biochemistry of the reproductive organs. After CcMtE treatment, significant decreases (p≤.001) were observed in the number of testicular cells (i.e., spermatogonia, primary spermatocytes [preleptotene and pachytene], secondary spermatocytes and round spermatids); nonsignificant change was observed in the Sertoli cell count. The treatment had no effect on levels of serum protein, cholesterol, bilirubin, glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), blood urea and hematological indices. CONCLUSIONS: The present study shows that C. cyminum treatment resulted in the inhibition of spermatogenesis and fertility without producing apparent toxic effects.


Subject(s)
Cuminum/chemistry , Fertility Agents, Male/analysis , Genitalia, Male/drug effects , Plant Extracts/pharmacology , Spermatogenesis/drug effects , Animals , Body Weight/drug effects , Fertility/drug effects , Fertility Agents, Male/pharmacology , Genitalia, Male/metabolism , Hematologic Tests , Male , Organ Size/drug effects , Rats , Rats, Wistar , Sperm Count , Sperm Motility/drug effects , Testosterone/blood
15.
Reprod Biol Endocrinol ; 9: 65, 2011 May 16.
Article in English | MEDLINE | ID: mdl-21575213

ABSTRACT

BACKGROUND: We describe the development of an electrochemical sensor array for monitoring the proliferation effects of cissus populnea plant extracts on TM4 Sertoli cells. METHODS: The proliferation activities of the extracts on Sertoli cells were studied using a high-throughput electrochemical sensor array (DOX-96) and the analytical sensor characteristics were compared with conventional colorimetric MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and fluorescence spectroscopy. RESULTS: This work shows that there is a definite positive trend in the proliferation effect of the extract of Cissus populnea on the TM4 Sertoli cells. All of the three techniques confirmed that the most effective concentration for the proliferation is 10 ppm. At this concentration, the proliferation effect was established around 120% for both DOX-96 and MTT techniques, whereas fluorescence assays showed a higher level (120-150%). DOX-96 showed a lower limit of detection (1.25 × 10(4) cells/ml); whereas the LOD recorded for both MTT and fluorescence techniques was 2.5 × 10(4) cells/ml. Visual examination of the cells by means of confocal fluorescence microscopy confirmed the proliferation of Sertoli cells as was determined using the MTT assay. This investigation provides a confident interpretation of the results and proved that the most effective concentration for the proliferation using Cissus populnea plant extract is 10 ppm. CONCLUSIONS: Overall, the DOX results compared well with the conventional methods of checking proliferation of cells. The fascinating feature of the sensor array is the ability to provide continuous proliferation experiments with no additional reagents including 96 simultaneous electrochemical experiments. The use of the DOX-96 could reduce a typical bioassay time by 20-fold. Thus the DOX-96 can be used as both a research tool and for practical cell culture monitoring.


Subject(s)
Biosensing Techniques/methods , Cell Proliferation/drug effects , Cissus/chemistry , Plant Extracts/pharmacology , Sertoli Cells/drug effects , Animals , Cells, Cultured , Electrochemistry/methods , Fertility Agents, Male/pharmacology , Fluorescent Antibody Technique/methods , Male , Rats , Sertoli Cells/physiology , Spermatogenesis/drug effects , Spermatogenesis/physiology , Tetrazolium Salts/pharmacology , Thiazoles/pharmacology
16.
J Basic Clin Physiol Pharmacol ; 22(1-2): 29-32, 2011 Jun 17.
Article in English | MEDLINE | ID: mdl-22865360

ABSTRACT

BACKGROUND: Cystic fibrosis (CF) is a hereditary disease caused by mutations of the gene encoding a channel protein CFTR, conducting Cl- and HCO3 - ions. The disease is characterized by disturbances in most physiological systems, and more than 95% of men are infertile. The mechanism underlying the etiology of CF is associated with an imbalance of fatty acids. It has been suggested that the function of the endocannabinoid system is also disturbed in CF, because endocannabinoids are derivatives of fatty acids. We assumed, therefore, that endocannabinoid activity, which plays an important role in fertility, is disrupted in CF and could be one of the causes of infertility. The aim of the present study was to test the hypothesis that stimulation of endocannabinoid receptors in infancy would normalize their function and prevent infertility in adulthood. METHODS: Knockout male mice (cftr-/-) were treated with tetrahydrocannabinol (THC), endocannabinoid receptors agonist, in infancy from days 7 until 28, daily. RESULTS: CF males treated with THC were fully fertile, producing offspring comparable by the number of litters and the number of pups with wild-type mice. CF males that were not treated with THC were completely infertile. CONCLUSIONS: The present study shows that (i) endocannabinoid function is impaired in CF mice, as evidenced by the regenerating effect of its stimulation on the fertility of otherwise infertile males, (ii) endocannabinoid system dysfunction is apparently the determining factor causing infertility in CF, and (iii) mild stimulation of the endocannabinoid system in infancy and adolescence appears to normalize many reproductive processes and thereby prevent infertility in CF males.


Subject(s)
Cystic Fibrosis/drug therapy , Dronabinol/pharmacology , Fertility Agents, Male/pharmacology , Fertility/drug effects , Infertility, Male/prevention & control , Animals , Birth Weight , Cannabinoid Receptor Agonists , Cannabinoid Receptor Modulators/metabolism , Cystic Fibrosis/complications , Cystic Fibrosis/physiopathology , Disease Models, Animal , Female , Infertility, Male/etiology , Infertility, Male/physiopathology , Litter Size , Male , Mice , Mice, Inbred CFTR , Pregnancy , Receptors, Cannabinoid/metabolism
17.
Theriogenology ; 73(1): 1-10, 2010 Jan.
Article in English | MEDLINE | ID: mdl-19828187

ABSTRACT

The aim of the current study was to assess the effect of insulin-like growth factor-I (IGF-I; 100 ng/mL) on buffalo (Bubalus Bubalis) sperm functional parameters related to in vitro fertilization. The acrosin activity (the mean diameter of halo formation in micrometers) was significantly higher in the IGF-I group (14.17 +/- 1.51) compared with that in the control group (9.50+/-0.36) at 2h incubation. The mitochondrial membrane potential (per cent) was significantly higher in the IGF-I group compared with that in the control group at 30min (33.27+/-2.62 vs. 26.71+/-1.02), 60min (24.24+/-3.45 vs. 18.77+/-2.09), and 90min (22.86+/-3.02 vs. 16.92+/-1.24) incubation. The percentage of spermatozoa positive for sperm nuclear chromatin decondensation (NCD) differed significantly between the groups at 90 and 120min incubation. The comet length was significantly lower in the IGF-I group compared with that in the control group at 2h incubation. The percentage of fragmented DNA in the tail did not differ significantly between the groups at 2h incubation. The percentage of acrosomal-reacted spermatozoa did not differ significantly between the IGF-I and the control groups at 4h (41.12+/-6.44 vs. 43.53+/-5.05) incubation. The cleavage rate (per cent) was significantly higher in the IGF-I-treated group (56.73+/-3.70) compared with that in the control group (44.85+/-2.15). The current study suggests that the addition of IGF-I prevents deterioration of sperm functional parameters and fertility.


Subject(s)
Buffaloes/physiology , Fertility Agents, Male/pharmacology , Fertility/drug effects , Insulin-Like Growth Factor I/pharmacology , Spermatozoa/physiology , Acrosin/physiology , Acrosome Reaction/drug effects , Animals , Chromatin/drug effects , Male , Membrane Potential, Mitochondrial/drug effects , Mitochondrial Membranes/drug effects , Semen Analysis , Sperm-Ovum Interactions/drug effects , Spermatozoa/drug effects , Spermatozoa/ultrastructure
18.
Forsch Komplementmed ; 16(6): 373-80, 2009 Dec.
Article in English | MEDLINE | ID: mdl-20090350

ABSTRACT

This review summarizes the current state of knowledge on Lepidium meyenii (maca), a cruciferous plant (Brassicaceae family) which is cultivated exclusively at an altitude of 4,000-4,500 m in the Peruvian Central Andes. Maca is traditionally used for its nutritional and presumed medicinal properties. Over the past 20 years, interest in maca has increased in many parts of the world, and since 2005 maca is considered one of the seven Peruvian flag products. Maca is exported as powder, capsules, pills, flour, liquor, and extracts. There are different types of maca with differ-ent colors ranging from white to black. We have studied the pharmacological effects of 3 types; yellow, black, and red maca. Evidence from experimental studies indicates effects of maca on nutrition, fertility, memory, and mood. Black maca has better effects on sperm production than yellow maca which has only moderate effects. Red maca, however, has no effect on sperm production. However, red maca has been shown to reduce prostate size in rats in which prostate hyperplasia had been induced with testosterone enanthate; yellow maca has shown moderate effects here, whereas black maca has not shown any effects. Randomized clinical trials have shown that maca has favorable effects on energy and mood, may decrease anxiety and improve sexual desire. Maca has also been shown to improve sperm production, sperm motility, and semen volume. Serum levels of testosterone, estradiol, LH, FSH, and prolactin were not affected. The exact mechanisms of action are still unclear, but so far research clearly indicates that various bioactive constituents contribute to the clinical effects reported.


Subject(s)
Lepidium/chemistry , Medicine, Traditional , Affect/drug effects , Animals , Clinical Trials as Topic , Fertility Agents, Male/pharmacology , History, 16th Century , History, 18th Century , History, 20th Century , History, Ancient , Humans , Male , Medicine, Traditional/history , Memory/drug effects , Nutritive Value , Peru , Plant Extracts/pharmacology , Spermatozoa/drug effects
19.
Zhongguo Zhong Yao Za Zhi ; 31(11): 904-6, 2006 Jun.
Article in Chinese | MEDLINE | ID: mdl-17048630

ABSTRACT

OBJECTIVE: Discuss the functional mechanism of treating male infertility with Bushen Shengjing pill (BS). METHOD: Fill the SD rat with adenine so as to injure the germ cell which will produce the model rat with kidney yang deficiency. Measure the activity of SOD and NOS, the amount of NO and MDA in the testicle. RESULT: BS can promote the activity of SOD, reduce the activity of NOS and the amount of NO and MDA. CONCLUSION: BS has the function of protecting the germ cell which is closely related to the function that the pill can promote the activity of SOD, reduce the activity of NOS and the amount of NO and MDA.


Subject(s)
Drugs, Chinese Herbal/pharmacology , Kidney Diseases/metabolism , Nitric Oxide Synthase/metabolism , Nitric Oxide/metabolism , Yang Deficiency/metabolism , Adenine , Animals , Drug Combinations , Drugs, Chinese Herbal/isolation & purification , Fertility Agents, Male/pharmacology , Kidney Diseases/chemically induced , Male , Malondialdehyde/metabolism , Plants, Medicinal/chemistry , Random Allocation , Rats , Rats, Sprague-Dawley , Spermatogenesis/drug effects , Superoxide Dismutase/metabolism , Testis/metabolism , Yang Deficiency/chemically induced
20.
Fitoterapia ; 77(6): 435-8, 2006 Sep.
Article in English | MEDLINE | ID: mdl-16889907

ABSTRACT

The effects of Butea superba on the reproductive system in male Wistar rats were investigated. The animals were fed daily with the powdered crude drug suspended in distilled water by a gastric tube at the dose of 2, 25, 250 and 1250 mg/kg body weight for 8 weeks. Rats fed with 1 ml of distilled water were used as a negative control. The weights of all vital organs in all treated groups were not different from the control. The percentage weight ratios of body weights of seminal vesicles and prostate glands were not different from the control, except that the testis of the group fed with 1250 mg/kg was significantly different from the control and the other treated groups. In addition, the sperm counts in this group showed about 16% more than the control group. Hematology as well as the liver and kidney function of all treated groups showed no difference from the control. B. superba, drug at 250 mg/kg which was 100 times more than the Thai FDA recommended dose for humans appeared to be safe in rats. The crude drug has demonstrated an increase tendency on testis weight and sperm counts in rat. The information from the present study can be used to explain the Thai folklore application of this plant in Thailand.


Subject(s)
Butea , Fertility Agents, Male/pharmacology , Phytotherapy , Plant Extracts/pharmacology , Reproduction/drug effects , Administration, Oral , Animals , Dose-Response Relationship, Drug , Erectile Dysfunction/drug therapy , Fertility Agents, Male/administration & dosage , Fertility Agents, Male/therapeutic use , Male , Plant Extracts/administration & dosage , Plant Extracts/therapeutic use , Plant Roots , Rats , Rats, Wistar , Sperm Count , Testis/drug effects
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