Effects of Genistein Isoflavone (4',5',7-Trihydroxyisoflavone) and Dexamethasone on Functional Characteristics of Spermatozoa

Caudal epididymal spermatozoa were used to study the influence of genistein isoflavone and dexamethasone (dxm) on the functional characteristics of spermatozoa. The effects of genistein alone and in combination with dxm on sperm motility, sperm morphology, spontaneous acrosome reaction (AcR), and ionophore A23187-induced AcR were investigated. The FITC-PSA/Hoechst 33258 staining procedure was used to assess sperm cell viability and AcR status and thus to differentiate between true AcR and acrosome degeneration. The overall results indicated that (1) lower doses of genistein alone, or in combination with dxm, did not significantly influence sperm motility or sperm morphology; (2) ionophore A23187 induced AcR in rat spermatozoa; (3) there appeared to be no direct correlation between sperm motility and AcR, (4) higher doses of genistein, alone or in combination with dxm, significantly interfered with percentage sperm motility and caused significant detachment of sperm heads but did not cause morphological defects; and (5) higher doses of genistein caused significant decrease in sperm acrosome reactivity with long duration of exposure. In view of the fact that sperm capacitation and AcR are physiological prerequisites for successful fertilization of oocytes, the findings suggest that chronic exposure of spermatozoa to high doses of genistein could be associated with infertility problems through suppression/inhibition of AcR and sperm motility. Dexamethasone did not appear to influence the effect of genistein on the functionality of postspermatogenic spermatozoa.     

HYALURONIC ACID INCREASES MOTILITY/INTRACELLULAR CA 2+ CONCENTRATION IN HUMAN SPERM IN VITRO

This study investigated the mechanisms of the stimulatory effect of hyaluronic acid on motility in human sperm in vitro. A method, involving the measurement of forward progression through an agarose gel, was used to measure sperm motility quantitatively. Changes in intracellular Ca 2+ concentrations in sperm were detected using the fluorescent dye Fluo-3. The effects of hyaluronic acid (6.5, 65, 650 ng/mL) and nifedipine (32 nM) on sperm motility were investigated. The effects of hyaluronic acid, nifedipine (32 nM), A23187 (32 µM), and a monoclonal antibody to human CD44 (1 µg/mL) on changes in intracellular CA 2+ concentrations were investigated. Hyaluronic acid significantly ( p <. 008) stimulated sperm motility and this was partially inhibited by nifedipine.A23187 significantly ( p <. 005) increased intracellular CA 2+ concentrations. Hyaluronic acid significantly ( p <. 04) increased intracellular Ca 2+ concentrations and this was inhibited by nifedipine and a monoclonal antibody to human CD44. Hyaluronic acid stimulated human sperm motility by increasing intracellular Ca 2+ concentration, partially via an influx of extracellular Ca 2+.     

A comparison of the effects of nonoxynol-9 and chlorhexidine on sperm motility

The effects of Nonoxynol-9 and chlorhexidine on sperm motility were compared using the objective Transmembrane Migration Ratio method. These agents were found to be of similar potency in inhibiting sperm motility. The concentrations which reduced sperm motility by 50% (EC50) were 0.205 mg/ml for Nonoxynol-9 and 0.215 mg/ml for chlorhexidine. The implications of these findings in relation to the comparison of the effects of drugs on sperm motility and the development of new contraceptive agents are discussed.     

Plasma Membrane Cytoskeletal Complex of the Mammalian Spermatozoa

Human and ram sperm were incubated in vitro in Tris buffer with (1) calcium ions, (2) ionophore A23187, (3) calcium plus A23187, (4) without any substance, as described in [13], and were reincubated in situ with the protein S-1 of heavy meromyosin (1 mg/ml). These spermatozoa were examined with the electron microscope to study and characterize the cytoskeletal structures after negative staining, freeze fracturing, and thin sectioning. The formation of the cytoskeletal complex, associated with the plasma membrane in the postacrosome region, appeared to be triggered by S-1 in those sperm that were earlier treated with the ionophore. The dynamic nature of actin present in the postacrosomal dense matrix becomes evident with the formation of a cytoskeletal basket in that region. The cytoskeletal complex may play an important role during the exocytotic acrosome reaction, helping to retain the shape of the sperm head and protect the genetic material until fertilization.     

Studies of the mechanism of action of gossypol as a male antifertility agent

Gossypol administered orally to male rats at a daily dose of 20 mg/kg body weight for 62 days caused infertility. There were changes in the epididymal epithelium and the sperm were severely damaged and immotile. The sperm head was often detached; other defects were abnormal mitochondria, absence of plasma membranes and axonemal and accessory fibres and a lower oxygen uptake. To study the effect of gossypol on the motor apparatus of sperm, ram sperm were demembranated with the detergent, Triton-X-100. Such sperm models can normally be reactivated with ATP but gossypol (2.5-12.5 microM) decreased reactivation and must have a direct effect on the axoneme. Gossypol also inhibited ram sperm adenyl cyclase which is essential for maintaining high levels of cAMP in sperm and, in turn, motility. Ram sperm adenyl cyclase required Mn2+ for activity and high Mn2+ concentrations protected the enzyme from gossypol inhibition. Electron spin resonance studies proved that gossypol chelated Mn2+ with the formation of a 2:1 complex.     

Effects of methylmercury on sperm and egg viability of two populations of killifish (fundulus heteroclitus)

Exposure of sperm of killifish (Fundulus heteroclitus) from a relatively clean area in Long Island (LI) to 0.01 mg/L methylmercury (meHg) in 15%o sea water caused significant reduction of fertilization success. However, exposure of killifish sperm from polluted Piles Creek (PC) to either 0.01 or 0.05 mg/L meHg in 15%o sea water prior to insemination had no effect on fertilization success. Exposure of LI killifish sperm to 0.05 mg/L meHg caused significant reduction in motility. However, PC killifish sperm showed no significant difference in motility between 0 and 0.05 mg/L meHg exposure. Exposure for 5 min to 0.05 mg/L meHg caused significant reduction in motility. These data indicate that meHg is less toxic to PC killifish sperm than LI killifish sperm. Exposure of PC and LI killifish sperm to 0.05 mg/L meHg for 15 min had no effect on sperm morphology. PC killifish sperm also showed higher (20 min) motility in 15%o sea water than LI killifish sperm (10 min). Exposure of PC and LI killifish eggs up to 25 min to 0.05 mg/L meHg prior to fertilization had no effect on fertilization success.     

轻质油（45℃馏分）蒸气的生殖毒性研究：Ⅰ.小鼠精子形态学检查

本文研究了小鼠吸入轻质油(45℃馏分)蒸气3000mg/m~3×6小时/天×30天条件下,小鼠精子形态学指标的改变。结果表明:亚急性吸入轻质油(45℃馏分)蒸气后,小鼠精子数目、精子活动率、精子活动度、精子畸形率与空白对照组比较,均无显著性差异(P>0.05)。本文讨论了精子形态学各指标与生殖能力的关系,指出精子数目减少,精子活动力下降,精子畸形率上升是生育力下降的原因之一。     

乙醇对雄性小鼠精子生成的影响

分析经口染毒乙醇３５天的雄性小鼠的精子数量、活动度、精子畸形率及睾丸细胞染色体畸变,研究乙醇对雄性小鼠精子生成的影响。结果表明,４０００ｍｇ／ｋｇ组和４５００ｍｇ／ｋｇ组小鼠精子活动度明显下降;３０００,４０００和４５００ｍｇ／ｋｇ组精子畸形率明显增高;４５００ｍｇ／ｋｇ组睾丸细胞染色体畸变率明显增高,与阴性对照组相比,上述指标差异均有显著性（Ｐ〈０．０１）。说明乙醇对小鼠的精子生成有一定损害作用     

乙烯利对小鼠生殖能力的影响

目的研究乙烯利对小鼠生殖能力的影响。方法选用乙烯利作为染毒药物,实验设阴性对照组（0mg／kg）、低剂量组（85mg／kg）、中剂量组（170mg／kg）和高剂量组（340mg／kg）,雄鼠经口染毒14d,于首次给药第15天起以1：1与雌鼠合笼。于合笼后第21天以颈椎脱臼处死雌鼠,剖腹检查确定受孕情况并计算生育仔鼠数。于首次给药第28天后用颈椎脱臼法处死雄鼠,观察乙烯利对雄鼠精子运动率的影响。结果高剂量组雌鼠受孕率与阴性对照组比较明显降低（P〈0．05）;与阴性对照组相比,高剂量组精子计数明显减少（P〈0．05）;各剂量组精子活动率均低于阴性对照组（P〈0．05）。结论乙烯利对雄性小鼠产生了一定的生殖毒作用。     

辛硫磷对大鼠精子生成量和精子运动能力的影响

为探讨有机磷杀虫剂辛硫的雄性生殖毒性,运用动物实验研究了不同剂量辛硫磷径口染毒大鼠６０天对精子生成量和精子运动能力的影响。采用计算机辅助精子分析仪分析精了轨迹。结果显示：辛硫磷２４．５和７３．５ｍｇ／ｋｂＢＷ染毒组,大鼠睾丸精子生成量低于对照组。     

甲胺磷对小鼠精子质量的影响

目的：研究甲胺磷对小鼠精子质量的影响。方法：用不同剂量（0．5,3．75,5．0,7.5mg/kg）的甲胺磷经腹腔注射染毒BALB／C雄性小鼠,4周后对小鼠精子数量,精子活率和精子形态进行分析。结果：染毒组小鼠精子畸形率与对照组相比显著增加,除0．5mg/kg剂量组外,其他剂量组小鼠精子活率显著下降,与对照组相比有统计学意义,5．0和7．5mg/kg剂量组小鼠精子数量明显低于对照组,结论：甲胺磷对小鼠精子有一定毒性。     

Fucoidin inhibits the zona pellucida-induced acrosome reaction in human spermatozoa

We recently reported that fucoidin (a polymer of predominantly sulfated L-fucose) significantly inhibits tight binding of human sperm to the human zona pellucida in vitro and that several oligosaccharides obtained after acid hydrolysis possess sperm-zona pellucida binding inhibitory activity equal to the original fucoidin. This inhibition may be specific to sperm-zona interactions or may be the consequence of the interruption of capacitation, a series of biochemical and physiological events leading to final sperm maturation, that must occur for successful fertilization. Completion of capacitation is most often determined by assessing two end-points of the process: acquisition of hyperactivated motility and ability to complete the acrosome reaction. Here, we examined the effects of fucoidin on these two end-points of capacitation in vitro. Fucoidin did not affect the proportion of sperm with hyperstimulated motility. Neither did fucoidin cause an increase in sperm that had spontaneously acrosome-reacted at 4.5 hours compared to controls as evaluated by indirect immunofluorescence using the acrosomal marker, monoclonal antibody, T-6. Comparable percentages of sperm had completed the acrosome reaction when exogenously stimulated by calcium ionophore A23187 with and without the addition of fucoidin. However, in the presence of fucoidin, stimulation of the acrosome reaction by acid solubilized human zonae pellucidae was significantly inhibited. These data indicate that fucoidin does not impede the normal progression of capacitation. These results provide strong evidence to support the hypothesis is that the inhibitory effect of fucoidin is at the level of the sperm membrane since inhibition can be bypassed by increasing intracellular calcium directly with a calcium ionophore.     

Improvement in human semen quality after oral supplementation of vitamin C

This study was carried out to monitor the effect of oral supplementation of vitamin C on various semen parameters in oligospermic, infertile, otherwise healthy individuals. Various semen parameters, including sperm motility, sperm count, and sperm morphology, were studied before and after the vitamin C treatment. A total of 13 infertile patients were included. Their ages ranged between 25 and 35 years. They had no genital infection or varicocele. Physical examination and other routine laboratory investigations were normal. General semen analysis revealed oligozoospermia (mean sperm count was 14.3 +/- 7.38 x 10(6) sperms/mL, mean sperm with normal morphology was 43 +/- 7.87%, and mean sperm motility was 31.2 +/- 9.61%). Testicular biopsy was not done. These patients received in an open trial of 1,000 mg of vitamin C twice daily for a maximum of 2 months. Results showed that the mean sperm count was increased to 32.8 +/- 10.3 x 10(6) sperms/mL (P < .001) after 2 months of vitamin C intake. The mean sperm motility was increased significantly to 60.1 +/- 8.47% (P < .001), and mean sperms with normal morphology increased significantly to 66.7 +/- 4.77% (P < .001). This study showed that vitamin C supplementation in infertile men might improve sperm count, sperm motility, and sperm morphology and might have a place as an additional supplement to improve the semen quality towards conception.     

Spermicidal activity of Azadirachta indica (neem) leaf extract

The present study was carried out to evaluate the effective concentration of aqueous extract of old and tender Azadirachta indica (neem) leaves to immobilize and kill 100% human spermatozoa within 20 s. Sander-Cramer test was used to study the spermicidal activity of neem leaf extract. Under the test conditions, minimum effective spermicidal concentrations for tender and old leaf extracts were 2.91 +/- 0.669 mg/million sperm and 2.75 +/- 0.754 mg/million sperm, respectively. The effect of extracts on morphology and viability of sperm was also studied and no change was observed in morphology of head, mid-piece and tail and no viable sperm seen. The leaf extracts were found to be water soluble and carbohydrate in nature. The effect of different concentrations of extracts (old and tender) on percentage motility of the sperm was also studied. With an increase in concentration, there is a linear decrease in percentage motility, becoming zero at a 3-mg dose within 20 s.     

Investigations on the use of testosterone oenanthate as a male contraceptive agent

The effect of exogenous testosterone on male gonadal function was in vestigated by applying a practical dose schedule of weekly injections of testosterone enanthate. 7 healthy young males received 22 intramuscular injections of 250 mg testosterone enanthate once a week for a period of 21 weeks. Mean sperm counts were reduced from a pretreatment value of 73 million sperm/ml to below 3 million sperm/ml within 9 weeks of testosterone administration and remained so during the whole treatment period. A marked recovery of spermatogenesis did not occur before 13 weeks after testosterone withdrawal, when the mean sperm count rose to 63 million sperm/ml. Changes of the mean sperm motility and percentage of normal sperm morphology generally paralleled those of the mean sperm concentration, whereas the mean semen volume, libido, potency, and secondary sex characteristics remained unaffected. A consistently azoospermic semen was not developed by any individual. This and the fact that sperm counts in 3 out of 7 males still showed some depression of spermatogenesis at the end of the observation period of 28 weeks after testosterone withdrawal indicate that testosterone enanthate admin istered in the described dose schedule is not a safe male contraceptive agent. However, further research on the use of hormones for the control of fertility in men seems warranted.     

十一酸睾酮对人精浆蛋白的影响

本文采用SDS—PAGE法,测定7例健康育龄男子在肌肉注射十一酸睾酮(TU)前,用药后l、2、3个月及停药后精子计数恢复正常时的精浆蛋白浓度和组份变化,并结合血清测定激素水平及常规精液分析,以探讨TU对上述指标的影响及其相互关系。结果显示:用药早期精浆总蛋白浓度开始逐步降低,在停药后精子计数恢复正常时仍未达到正常水平。精浆总蛋白浓度的变化与精子活动率、前向运动率及LH呈正相关(r值分别为0.8975、0.7776、0.6406,P<0.01或0.05)。精浆蛋白组份也发生变化,其中51KD、37KD的蛋白条带变化最为明显,其变化除与精子活动率相关外(r=0.9094,P<0.01),其中51KD蛋白条带的变化还与精子前向运动率相关(r=0.7660,P<0.05)。上述结果提示TU可能在抑制生精之前即通过改变精浆蛋白而影响精子的功能,说明单用睾酮(T)的避孕机理是多因素的。     

农达对小鼠生精作用的影响

目的探讨农达(roundup,活性成分为草甘膦)对小鼠生精作用的影响。方法将24只成年SPF级雄性Balb/c小鼠随机分为阴性对照(蒸馏水)组和20、200、2 000 mg/kg农达染毒组,每组6只。采用经口灌胃方式进行染毒,染毒容量为20 ml/kg,每天1次,连续染毒30 d。测定小鼠精子数量、精子活动率和血清中睾酮(T)浓度及睾丸生精细胞的凋亡情况。结果与阴性对照组相比,200、2 000 mg/kg农达染毒组小鼠精子数量减少、精子活动度降低;各剂量农达染毒组小鼠血清睾酮浓度均降低,而睾丸生精细胞的凋亡率均升高,差异均有统计学意义(P0.05)。结论在本实验染毒时间和剂量范围内,农达对雄性Balb/c小鼠的生精功能有一定的毒性作用。     

Induction of Oxidative Stress in the Epididymis of Rats After Subchronic Exposure to Epichlorohydrin

This study investigated the effects of epichlorohydrin (ECH) on spermatogenesis and antioxidant system in rats. An increase in the incidence of clinical signs, gross pathology and histopathology findings in the epididymidis, and sperm abnormalities and a decrease in the testicular spermatid counts, epididymal sperm counts, and sperm motility were observed at 30 mg/kg/day. Oxidative stress in the epididymal tissue was detected at ≥3.3 mg/kg/day. The results show that graded doses of ECH elicit depletion of antioxidant defense system and that the adverse effects on male reproductive function in ECH-treated rats may be due to the induction of oxidative stress.     

精索静脉曲张不育患者8-羟基脱氧鸟苷水平的检测分析

目的:检测人血清及精浆中8-羟基脱氧鸟苷(8-OHdG)水平,探讨男性精索静脉曲张(VC)患者体内氧化损伤情况。方法:对41例VC不育患者、33例正常生育者进行精液分析,用8-OHdG酶联免疫试剂盒检测两组对象血清及精浆中8-OHdG水平,采用Wilcoxon两样本比较法对数据进行统计分析。结果:正常生育组的精子浓度、前向活动率、存活率均高于VC不育组(P<0.05),血清8-OHdG水平(中位数34.83ng/ml)低于VC组(中位数76.87ng/ml,P<0.05),精浆8-OHdG水平(中位数18.51ng/ml)低于VC组(中位数22.19ng/ml,P<0.05)。血清中8-OHdG水平高于精浆(P<0.05)。血清8-OHdG水平与精子存活率、前向活动率呈负相关(γ=-0.293,P<0.05;γ=-0.243,P<0.05);精浆8-OHdG水平与精子存活率、前向活动率呈负相关(γ=-0.327,P<0.05;γ=-0.275,P<0.05);血清及精浆中8-OHdG水平与精子浓度、正常形态率不相关。结论:VC患者血清和精浆中8-OHdG水平高于正常生育组,表明VC患者可能存在DNA氧化损伤,导致精子活力下降。     

Curcumin: a potential vaginal contraceptive

The purpose of this investigation was to evaluate the sperm-immobilizing effects of curcumin, a plant-derived diferuloylmethane compound. Washed human healthy sperm were suspended in Ham's F10 and exposed to varying concentrations of curcumin. Sperm motility was evaluated and changes in sperm mitochondrial transmembrane potential (MTP) was quantified by flow cytometry. Incubation of normal human sperm with curcumin resulted in a dose- and time-dependent loss of sperm motility. At lower concentrations (30 g/mL), curcumin produced a significant (20%) decrease in sperm motility within 30 min without significant effects on sperm viability. An instantaneous (>50%) loss of sperm motility was observed with higher concentrations (300 g/mL) of curcumin and a total loss of sperm motility was achieved within 60 min. A significant reduction in sperm MTP was found with all doses of curcumin tested. Our results indicate that curcumin has a selective sperm-immobilizing effect, in addition to a previously studied anti-HIV property. This compound may have potential clinical applications as a novel intravaginal spermicidal agent for contraception and HIV prevention.