Effects of lithium and haloperidol on human sperm motility in-vitro.

Two psychotropic drugs, lithium and haloperidol, were evaluated for their in-vitro effects on sperm motility using a transmembrane migration method. Sperm motility was measured either immediately after semen had been mixed with the drug or after a 2 h incubation period at 37 degrees C. Lithium inhibited human sperm motility in a dose-dependent manner with an EC50 of 10 mM when the semen-lithium mixture had been incubated. Sperm motility was increased to 127% of control when semen had been incubated with 0.027 microM haloperidol; this concentration was within the therapeutic range.     

吸入麻醉剂浓度及作用时间对人离体精子活率的影响

目的:研究吸入麻醉剂浓度及作用时间对人离体精子活率的影响。方法:选择人精液20份,上游法优化处理后随机分为异氟醚实验组和七氟醚实验组,各10份。分别观测5个时间点(0.5 h、1 h、2 h3、h4、h)及5个浓度水平(0、1.4%、2.8%、4.2%、5.6%)异氟醚和相似浓度七氟醚对精子活率的影响,精子运动功能采用计算机辅助精子分析系统分析。结果:1.4%~5.6%异氟醚作用于精子0.5~4 h后,精子活率显著升高,其变化趋势符合S型曲线,相同浓度异氟醚组,精子活率随放置时间的延长成直线下降。类似浓度七氟醚对精子活率无显著影响。结论:临床浓度异氟醚作用于人离体精子,可以显著升高精子活率。类似浓度七氟醚对精子活率无显著影响。     

Effects of chlorpromazine and other drugs acting on the central nervous system on human sperm motility

Summary The in vitro effects of chlorpromazine, diazepam, phenytoin and phenobarbitone on human sperm motility were investigated. Only chlorpromazine inhibits human sperm motility and the concentration which decreases sperm motility to 50% of control is 0.22 mM. Caffeine can shift the dose-response curve of chlorpromazine-inhibited sperm motility to right. We support the hypothesis that chlorpromazine acts on the cellular membrane but consider the inhibition of sperm motility an unlikely cause of decreased fertility in chlorpromazine treated patients.     

Effects of lithium on calmodulin-stimulated adenylate cyclase activity in cortical membranes from rat brain

The influence of lithium on calmodulin-stimulated adenylate cyclase activity has been studied in vitro and after chronic treatment. Chronic lithium treatment decreased calcium-calmodulin-stimulated adenylate cyclase activity in rat cortical membranes, while no effect was observed on GTP-stimulated activity. Lithium in vitro inhibited adenylate cyclase activity stimulated by isoprenaline, GTP or calcium-calmodulin. Calcium-calmodulin-stimulated activity was more sensitive to lithium (2 mM) than isoprenaline- and GTP-stimulated activities (5 mM) and activities by these agents combined. Lithium had no effect on the unstimulated enzyme activity. The inhibitory effect of lithium in vitro on calcium-calmodulin-stimulated adenylate cyclase activity was antagonized by magnesium. The inhibition induced by lithium in vitro on the GTP-stimulated adenylate cyclase activity was increased by substituting manganese for magnesium in the assay media. Furthermore, the manganese-stimulated activity was also reduced by lithium. The latter effect was not observed in calmodulin-depleted membranes, but the inhibitory effect of lithium could be restored by addition of exogenous calmodulin. The present results suggest that lithium might influence the interaction of calmodulin with the enzyme and/or interfere with the divalent cation site(s) on the adenylate cyclase system.     

槟榔碱对人体外精子运动能力的影响

目的:研究槟榔碱(Ar)对人体外精子运动能力的影响。方法:优选50例正常男性精液与3组不同浓度(10、50、100μg·mL-1)Ar溶液共同孵育,以精子优选液为对照组,在共同孵育0.5、1、2h后用计算机辅助精子分析(CASA)系统对精子活率(Mot)、(a+b)级前向运动精子百分率([a+b)PM]、曲线运动速度(VCL)、直线运动速度(VSL)进行检测。结果:精液与10μg·mL-1Ar溶液共同孵育1h后的Mot与对照组比较有显著性差异(P<0.01),2h后Mot、VCL与对照组比较均有显著性差异(P<0.01);精液与50、100μg·mL-1Ar共同孵育0.5、1、2h后的Mot、(a+b)PM、VCL、VSL与对照组比较均有显著性差异(P<0.01或P<0.05),表明Ar浓度越高,作用时间越长,精子运动能力越低。结论:Ar能降低正常男性体外精子运动能力,其毒性与浓度时间成正比。     

Influence of lithium on cyclic AMP accumulation in isolated rat fat cells.

Lithium inhibits in vitro as well as in vivo several hormone-stimulated adenylate cyclases. The aim of this study was to investigate the mechanism by which lithium inhibits adenylate cyclase in vitro. It was found that lithium inhibited both the norepinephrine- and the glucagon-induced cAMP accumulation in rat fat cells at lithium concentrations above 10 mM. The basal cAMP content was unaffected even at 40 mM of lithium. The inhibitory action was time-dependent and reversible, indicating an intracellular site of action. Lithium inhibited both norepinephrine- and glucagon-stimulated cAMP accumulation in a mainly non-competitive way, but the inhibitory effect decreased with increasing hormone concentrations. In accordance, lithium and propranolol had a supraadditive effect on norepinephrine-induced cAMP accumulation. It is suggested that lithium affects both the hormone-receptor binding as well as the transfer of the hormonal stimulus by an intracellular site of action.     

不同有机溶剂对人精子体外运动参数的影响比较

目的研究常用有机溶剂对正常男性精子体外运动活力的影响。方法取正常男性精子,采用上游优化法处理,制备成精子悬液并分成4组,前3组分别加入二甲亚砜,使二甲亚砜终浓度分别为0.5%,1%和2%,pH值均为7.5,渗透压为290 mosm.(kg.H2O-1),D组加入等量Ham’s F10培养液作对照。分别于孵育15,30,45和60 m in后采用CASA进行分析。参照上述方法,考察有机溶剂甘油、聚乙二醇、吐温80对精子体外运动活力的影响。甘油终浓度分别调至1%,5%和10%,聚乙二醇终浓度分别调至1%,5%和10%,吐温80终浓度分别调至1%,5%和10%。结果2%二甲亚砜作用于精子60 m in能导致精子全部死亡,10%甘油作用60 m in后对精子运动参数抑制率达73.8%,10%聚乙二醇400作用于人精子60 m in可致其全部死亡,10%吐温80作用60 m in后对精子运动参数抑制率达95.5%。加入不同浓度二甲亚砜、甘油、聚乙二醇400和吐温80的各组前向运动精子百分率、曲线运动速度、直线运动速度和平均路径速度均较空白对照组显著降低(均P<0.05)。结论高浓度有机溶剂体外可抑制正常男性精子存活率和运动活力。     

Calcium antagonists stimulate sperm motility in ejaculated human semen

Three calcium antagonists, diltiazem, flunarizine and verapamil stimulated human sperm motility in vitro. Among them, diltiazem induced the largest amplitude of motility increase. The concentration-response curve of diltiazem was similar in shape to those of calcium chelators. EGTA, a calcium chelator, potentiated the stimulatory effect of diltiazem while A23187, a calcium ionophore, antagonized it. These observations supported our previous hypothesis that an increased concentration of calcium ion was detrimental to human sperm motility. Whether calcium antagonists could be used in the treatment of subfertile patients awaits further investigation.     

Effects of fenvalerate and cypermethrin on rat sperm motility patterns in vitro as measured by computer-assisted sperm analysis

Fenvalerate and cypermethrin were reported to impair male reproductive function, inducing significant reductions in epididymal sperm count. Further, fenvalerate was shown to reduce sperm motility. However, it is not clear whether fenvalerate and cypermethrin might impact sperm motility directly or indirectly by affecting spermatogenesis via interaction with androgens or their receptors. In this study, sperm suspensions were treated with fenvalerate and cypermethrin, respectively, at various concentrations (0, 1, 4, 16, or 64 micromol/L) for various times (1, 2, or 4 h). The motility parameters of sperm treated with these two insecticides were analyzed with a computer-assisted sperm analysis (CASA) system. The differential effects of fenvalerate and cypermethrin on rat sperm motility patterns in vitro were also compared. Our study revealed that fenvalerate and cypermethrin reduced sperm motility in vitro in a concentration- and time-dependent manner. Cypermethrin exerted a greater effect on sperm motility in comparison to fenvalerate. These results provided evidence that fenvalerate and cypermethrin directly influence mature rat sperm motility.     

Subchronic supplementation of lithium carbonate induces reproductive system toxicity in male rat

Lithium is frequently used as an effective drug for the treatment of several psychiatric disorders in human. This alkali element and its salt, at its higher doses, may lead to various side effects or has several toxic effects after prolonged therapeutic use. To test this hypothesis, the present study was designed to investigate the adverse effect of subchronic exposure of lithium carbonate on reproductive organs of male rat. Rats were exposed to lithium carbonate at doses of 500, 800, 1100 mg/kg of diet for 90 days. The weight of reproductive organs, histology of testis, epididymis, seminal vesicle, prostate, testicular interstitial fluid volume (IFV), testosterone level, sperm morphology and fertility index were analyzed. Treatment with higher doses of lithium carbonate (i.e. 800, 1100 mg/kg diet) significantly reduced testes, epididymis and accessory sex organs weights, whereas, lower dose (500 mg/kg diet) did not show any untoward effect. Similarly, the sperm number from cauda epididymis and daily sperm production was significantly decreased with higher doses of lithium carbonate. The serum testosterone levels and IFVs were also reduced significantly. Seminal vesicle and prostate secretions were completely blocked and spermatozoa were not seen in the lumen of epididymis and vas deference. Histological studies have revealed that lithium carbonate (1100 mg/kg) caused degeneration of spermatogenic cells and vacuolization of sertoli cells cytoplasm in the testis. The sperm transit rate and production of abnormal spermatozoa were significantly (P<0.01) increased. When the lithium carbonate-treated males were mated with normal cyclic females, the fertility index declined to 50% even after 30 days of withdrawal of lithium carbonate treatment. These results clearly suggest that subchronic exposure of lithium carbonate promote reproductive system toxicity and reduces fertility of male rats.     

Lithium regulates protein tyrosine phosphatase activity in vitro and in vivo

RATIONALE: Lithium has been shown to regulate multiple intracellular signaling pathways by affecting various protein kinases. However, the counterpart of protein kinases, i.e., protein phosphatases may play an important role in lithium-regulated cellular signaling and functions. OBJECTIVES: The present work was designed to test the effect of lithium on protein phosphatases in vitro and in vivo. METHODS: PC12 cells were used as an in vitro model to characterize the effect of lithium on protein phosphatase activity. Rats treated with a lithium-containing diet were used to examine the in vivo effect of the drug on brain protein phosphatase activity.RESULTS. Lithium stimulated protein tyrosine phosphatase (PTPase) activity in a dose- and time-dependent manner in PC12 cells. A maximal stimulation of 87% was observed after 6 h of incubation with 3 mM LiCl. In contrast, protein serine phosphatase (PSPase) activity was not changed by lithium. The stimulatory effect on PTPase was not due to a direct action of the ion on the enzymes, but its selectivity was noted since treatment of cells with other monovalent cations exhibited no effect on PTPase activity. Lithium appeared to target specific PTPase(s) as it stimulated membrane-associated PTPase activity without affecting cytosolic or nuclear enzymatic activities. Moreover, the stimulation of PTPase activity in PC12 cells by lithium is independent of de novo protein synthesis. In the rat, 3 weeks of lithium treatment significantly elevated PTPase activity in hippocampus, striatum and cortex. CONCLUSION: The present findings provide the first evidence that lithium treatment selectively increases membrane-associated PTPase activity and suggest that this action may contribute to the pharmacotherapeutic actions of lithium.     

In vitro effects of arecoline on sperm motility and cyclooxygenase-2 expression

Semen samples were obtained from 30 volunteers who had never consumed betel quid. Swim-up spermatozoa from the 30 seminal samples of non-betel quid chewers and also non-smokers, usually not exposed to passive smoking, were treated in vitro with arecoline at different concentrations to evaluate the action of these drugs on sperm motility. Highly motile sperms were collected and divided into 5 equal fractions. Four fractions were supplemented with various concentrations of arecoline and one as control. The study was carried out at time 0 and +1, +2, +3 and +4 hr of incubation. Sperm cells were also extracted and blotted with COX-2 antibody after arecoline treatment after 4 hr incubation. The sperm motility parameters, i.e., motility, average path velocity, curvilinear velocity, straight-line velocity and linearity, were significantly decreased after arecoline treatment. In vitro, arecoline induces the COX-2 expression of sperm cells in a dose-dependent manner. This is the first report to demonstrate that arecoline may mediate COX-2 expression in human sperms, resulting in inflammation response. This situation may act on the structure responsible for the flagellar motion and cause the reduction of sperm motility.     

Effects of permethrin,cypermethrin and 3-phenoxybenzoic acid on rat sperm motility in vitro evaluated with computer-assisted sperm analysis

Pyrethroid pesticides, produced and used worldwide, have been reported to impair male reproductive function by reducing sperm count and sperm motility. They are divided into two types: type I pyrethroids including permethrin, etc. and type II pyrethroids including cypermethrin, fenvalerate, cyfluthrin, etc. Our previous study showed that fenvalerate and cypermethrin could reduce sperm motility in vitro. However, it is not clear whether permethrin and 3-phenoxybenzoic acid (3-PBA, the major metabolite of pyrethroids) affect sperm motility directly or indirectly by affecting spermatogenesis via interaction with androgens and/or their receptors. In this study, rat sperm suspensions were treated respectively with permethrin, cypermethrin and 3-PBA, at various concentrations (0, 1, 4, 16, or 64 mmol/L) for various times (1, 2, or 4 h). The motility parameters of sperm were analyzed with a computer-assisted sperm analysis (CASA) system. The differential effects of permethrin and cypermethrin on sperm motility patterns in vitro were also compared. Our study revealed that permethrin and cypermethrin could reduce sperm motility in vitro in a concentration- and time-dependent manner. Marked differences between the two pyrethroids were not found in this study. Moreover, 3-PBA did not reduce sperm motility directly at all concentrations and treatment periods. These results provide further evidence that permethrin and cypermethrin can directly affect mature rat sperm motility.     

Adenosine Stimulates Human Sperm Motility via A2 Receptors

Abstract— The effects of adenosine and its analogues on human sperm motility were studied using a transmembrane migration method. Specific binding sites for adenosine in human sperm were also investigated. Adenosine and 5′-N-ethylcarboxamidoadenosine (NECA) stimulated human sperm motility with similar efficacies and the maximal amplitudes of motility increases were both about 70%. 3,7-Dimethyl-1-propargylxanthine (DMPX), a potent A₂ antagonist, competitively antagonized NECA-induced motility stimulation. Successively higher concentrations of DMPX shifted the dose-response curve of NECA to the right in a nearly parallel fashion. Dipyridamole, an inhibitor of adenosine uptake, does not reduce the ability of adenosine to stimulate human sperm motility. In radioligand-binding studies, adenosine A₁ selective analogues, cyclopentyl-1,3-dipropylxanthine and 1-methyl-2-phenylethyl adenosine, have little competitive effect on [³H]NECA binding in human sperm membrane. These results provide evidence that adenosine enhances human sperm motility via adenosine A₂ receptors on the surface of sperm membranes.     

Effect of two insecticides and two herbicides on the porcine sperm motility patterns using computer-assisted semen analysis (CASA) in vitro

The recent decline in sperm concentration observed in men has developed over a short period of time, suggesting that it could be the result of environmental factors. The present study has evaluated the effects of insecticides Malathion and Diazinon, and herbicides Atrazine and Fenoxaprop-Ethyl on porcine sperm viability and motility patterns in vitro using the eosin-nigrosin staining and a computer-assisted semen analyzer (CASA), respectively. Malathion and Fenoxaprop-Ethyl exerted more deleterious effects than Diazinon and Atrazine. Progressive sperm motility was strongly affected whereas the effect on sperm viability was less pronounced. This suggests that a reduction of sperm motility is not necessarily the result of sperm death. Since sperm motility is dependent on energy metabolism the mechanism of action of these pesticides might be mediated at the level of the mitochondrion, producing a delay in motility and eventual cell death.     

The effects of lithium in vitro and ex vivo on adenylate cyclase in brain are exerted by distinct mechanisms

The effects of lithium on basal and forskolin-stimulated activity of adenylate cyclase in membrane preparations from cerebral cortex of the rat have been studied. Chronic treatment with lithium, yielding a level of lithium in serum of 0.71 +/- 0.18 mmol/l, reduced forskolin-stimulated activity in total homogenates but exerted no effect on the basal activity. Lithium in vitro, at 2 and 10 mM, did not influence the basal enzyme activity in membranes from either control or lithium-treated animals. The sensitivity of forskolin-stimulated adenylate cyclase to lithium in vitro was unaltered after chronic treatment and the in vitro and ex vivo effects of lithium on this parameter were additive. The inhibitory ex vivo effect of lithium was not antagonized by increasing concentrations of magnesium and the inhibitory effect of lithium ex vivo was still persistent after washing of the membranes. The present results indicate that lithium exerts its ex vivo effect on the activated cyclase, independently of the in vitro effect. Both effects may, however, contribute to the in vivo effect of lithium during chronic treatment.     

苯扎溴胺对精子运动能力的影响

目的：运用计算机辅助精子质量检测系统测定表面活性剂苯扎溴胺对精子运动的影响。方法：新鲜精液液化后，经离心洗涤与不同浓度的苯扎溴胺混匀，30s后滴片，置计算机辅助精子质量检测系统分析。结果：经苯扎溴胺处理的精子的活率和精子活力随着药物剂量的增加而逐渐降低，至0．25mg/ml浓度时，无活动精子，与对照组相比，差异有显著性（P＜0．01）；其直线运动速率VSL、曲线运动速率VCL及平均路径速率VAP，随剂量的增加逐渐降低，且各剂量组与对照组相比，差异均有显著性（P＜0．01）；侧摆幅度ALH与对照组相比，差异无显著性，P＞0．05；而用药组精子的直线笥LIN和前向性STR百分率均低于对照组，但是参数的变化并不规则。结论：苯扎溴胺可显著地影响精子的运动能力，从而使受精能力下降，作为外用避孕药使用是可行的。     

Lithium salts in AIDS and AIDS-related dementia

Agents currently in use against human immunodeficiency virus (HIV) infection, including the prospects of a vaccine, have focused primarily on destroying or disabling the virus. This strategy is hampered by the ephemeral nature of the viral genome. Lithium salts are widely used in psychiatry as mood-stabilizing agents. Studies have revealed, however, that the lithium ion also has significant granulopoietic actions, as well as regulatory effects on select cytokines that enable it to boost the body's natural defense against viral infections, specifically DNA viruses. Moreover, case reports in acquired immunodeficiency syndrome (AIDS) patients, as well as animal studies using related immunodeficiency viruses, provide support for a novel therapeutic role for lithium salts in the treatment of HIV infection. Furthermore, recent studies have revealed an important additional benefit of the lithium ion. In both in vitro and human studies, lithium has been found to increase the synthesis of neuroprotective proteins and to exert possible neurotrophic effects in the human brain. These properties hold great promise in the treatment of AIDS-related neurological deficits such as dementia. This paper reviews the mechanisms involved in and the clinical promise for lithium salts in the treatment of AIDS and AIDS-related dementia.     

Lithium: ADH antagonism and ADH independent action in rats with diabetes insipidus

Treatment with lithium chloride (2 mmole/kg × 24 hr) caused a gradual increase in water intake, urine output, sodium and potassium excretion of rats; Na/K in the urine also increased. Lithium administration caused partial depletion of hypophyseal vasopressin and did not prevent the depletion caused by hypertonic stimulation. Sodium administration also caused partial depletion of hypophyseal vasopressin but had no significant effect on water intake or urine output. The inhibition of water intake and urine output by administration of ADH to rats with hereditary diabetes insipidus was antagonized by lithium chloride (2 mmole/kg); the dose of ADH necessary to cause 50% inhibition was increased by lithium more than 5-fold. Lithium did not inhibit in vitro kidney adenylate cyclase from rats with diabetes insipidus while it produced significant inhibition in kidney adenylate cyclase from control rats. Lithium injected into untreated rats with hereditary diabetes insipidus further increased their water intake, urine output and decreased their urine osmolality. Therefore, lithium also affected water balance through a mechanism unrelated to ADH. In rats with D.I., lithium did not increase sodium excretion but decreased potassium excretion, thus Na/K in the urine was increased. The possible interaction of lithium with ADH and with aldosterone is discussed.     

Ginsenoside Re increases fertile and asthenozoospermic infertile human sperm motility by induction of nitric oxide synthase

We investigated the effects of Ginsenoside R(e) on human sperm motility in fertile and asthenozoospermic infertile individuals in vitro and the mechanism by which the Ginsenosides play their roles. The semen samples were obtained from 10 fertile volunteers and 10 asthenozoospermic infertile patients. Spermatozoa were separated by Percoll and incubated with 0, 1, 10 or 100 microM of Ginsenoside R(e). Total sperm motility and progressive motility were measured by computer-aided sperm analyzer (CASA). Nitric oxide synthase (NOS) activity was determined by the 3H-arginine to 3H-citrulline conversion assay, and the NOS protein was examined by the Western blot analysis. The production of sperm nitric oxide (NO) was detected using the Griess reaction. The results showed that Ginsenoside R(e) significantly enhanced both fertile and infertile sperm motility, NOS activity and NO production in a concentration-dependent manner. Sodium nitroprusside (SNP, 100 nM), a NO donor, mimicked the effects of Ginsenoside R(e). And pretreatment with a NOS inhibitor N(omega)-Nitro-L-arginine methyl ester (L-NAME, 100 microM) or a NO scavenger N-Acetyl-L-cysteine (LNAC, 1 mM) completely blocked the effects of Ginsenoside R(e). Data suggested that Ginsenoside R(e) is beneficial to sperm motility, and that induction of NOS to increase NO production may be involved in this benefit.