壬苯醇醚-9和氯苯胍亭作用于精子的比较

壬苯醇醚-9和双醋氯苯胍亭对精液中和宫颈粘液中精子活动力作用的比较。这两种化合物在精液中杀精作用相同,用0.5mg/ml的药物在3分钟内就可使精子丧失活动力。当这两种化合物扩散于宫颈粘液中时,粘液中存活精子状况是有差异的。在用0.1mg/ml氯苯胍亭处理后,立即发现精子穿透受到限制并丧失活动力,而用200mg/ml氯苯胍亭长期接触后的粘液中精子却正常存活。在精子穿透粘液前将两种化合物直接混合于粘液中,氯苯胍亭仍然制动精子,但壬苯醇醚-9的浓度在精液中有杀精作用而在粘液中却没有杀精作用。不过目前大多数化学性阴道避孕药是用壬苯醇醚-9。在性交前阴道表面活性避孕药作为泡沫剂,乳胶剂或宫托剂使用,杀精作用方式是引致精子体部     

油茶皂素不育剂对鼠类抗生育功能的实验研究

目的研究油茶皂素不育剂对鼠类的抗生育功能。方法测定不同浓度油茶皂素不育剂对雄性小鼠的体外杀精效果和体内抗生育能力。结果油茶皂素不育剂对雄性小鼠呈现出较优的剂量依赖性体外杀精活性,3 min时的最低杀精浓度为0.05 mg/ml,与人用杀精剂壬苯醇醚相当,比扁桃酸(0.1 mg/ml)和柠檬酸(0.25 mg/ml)低。与生理盐水对照组比较,125 mg/(kg.d)油茶皂素不育剂灌胃给药4周后,可以明显降低雄性小鼠的活精子数(92.1%)和精子活力(52.6%),可以显著减少雌性小鼠的妊娠率(64.0%)和活胎数(67.4%)。结论油茶皂素不育剂具有较强的体外杀精活性和体内抗生育功能,可进一步开发成体外杀精避孕药和环境友好型鼠类抗生育药剂。     

杀微生物避孕凝胶剂的抗生育和抗菌作用研究

目的:观察杀微生物避孕凝胶剂的体外杀精、家兔抗生育和抗菌作用。方法:杀精试验采用改良的Sander-Crammer方法;家兔抗生育试验将家兔随机分为高、中、低3个剂量组,分别阴道给予凝胶剂48mg/kg,24mg/kg和12mg/kg,给药后10min与雄兔进行交配,21d后解剖并计算避孕率;抗菌实验用体外试管法观察凝胶剂的抗金黄色葡萄球菌、大肠埃希菌和白色念珠菌的作用。结果:在20s内最低体外杀精浓度为0.5mg/ml;3个剂量组家兔避孕率分别为100%、80%和20%;对金黄色葡萄球菌、大肠埃希菌和白色念珠菌的最低杀菌浓度分别为0.05625%、0.225%和0.45%。结论:研制的凝胶剂有明显的抗生育作用和抗上述3种致病菌作用。     

壬苯醇醚—9外用避孕药药理和毒理研究

壬苯醇醚-9最低有效杀精子浓度0.2～0.25mg/ml。小鼠ip的LD_(50)为128.5±7.7mg/kg、大鼠阴道给药的LD_(50)>500mg/ml。33倍人用剂量亚急性试验给药组与空白对照组无明显差异(P>0.05)。100mg/ml浓度不抑制阴道乳酸杆菌。1mg/皿进行的Ames试验结果阴性。家兔阴道刺激试验结果总分值为6.63,在可接受范围。     

从无患子分离出的皂角甙与人的精液在体外混合后,在扫描电镜观察下,精子的形态学变化

最近,对阴道避孕药结合有效的杀精子剂这一发展又重视起来。据报道:许多药如表面活性剂,抗心律失常药,抗抑郁药和一些螯合剂均具有效的杀精子活性。在这些药中,去污剂如Nonoxynol-9的浓度为0.12mg/ml时,可影响精子的活动力。去污剂的杀精子作用是通过损伤精浆膜实现的。本文报道从无患子分离出的皂角甙的杀精子活性。本研究所采用的人体新鲜精液具有计数满意,前向活动力强,形态正常的特点。2ml精液与2ml无患子液(用盐水稀释,浓度分别为0.05%、0.1%、1.25%和5%)混合孵育1.5和10分钟。从同一标本中取2 ml精液仅与盐水孵育作为对照。然后在标     

洁泽2号体外杀精及大鼠阴道避孕实验

目的:观察中药复方药液洁泽1号(Jieze NO.1,JZ1)对外用杀精剂壬苯醇醚(N-9)的杀精增效作用及洁泽2号(Jieze NO.2,JZ2,JZ1与N-9混合制剂)凝胶大鼠阴道内用药的避孕(抗生育)效果。方法:采用改良的Sander-cramer方法观察JZ2中N-9的最低杀精浓度对正常人精子前向运动率、活动率和存活率的影响;大鼠阴道内给予JZ2药物凝胶后雌雄合笼,分别于交配后1 min、3 min、5 min、10 min观察阴道内精子活动情况,并观察各组避孕率。结果:N-9 20 s、3 min最低杀精浓度均为0.25 mg/ml,与洁泽1号混合(即JZ2)后,N-9最低杀精浓度分别为0.125 mg/ml、0.062 5 mg/ml。交配后1 min,各单纯N-9及JZ2凝胶组大鼠阴道内均未见活动精子。N-9(12%及以上浓度)、JZ2(含10%及以上浓度N-9)各组均有较高的避孕率,与自然对照组及空白凝胶组之间的差异均有极显著统计学意义(P<0.01)。JZ2(含14%、12%N-9)药物凝胶组抗生育率均可达100%,与14%N-9作用相当。JZ2(含10%N-9及8%N-9)抗生育率分别为90%、40%,与自然对照组之间有统计学差异。JZ2(含12%N-9和10%N-9)凝胶组对大鼠的避孕作用强于相同浓度的N-9组,且比较差异有统计学意义(P<0.05)。结论:由剂量-反应关系曲线可以看出JZ1对N-9的杀精作用具有协同作用,两药混合后(即JZ2)可以减少N-9的剂量;JZ2凝胶大鼠阴道内用药具有较好的抗生育作用,在相同避孕效果下可减少N-9的用量。     

白芸豆植物凝集素对不同物种精子的凝集作用与抗孕效果研究

目的:观察和比较自白芸豆中提取的植物凝集素(PHA)对人、小鼠、牛、山羊和兔精子的凝集作用和整体给药的避孕和抗早孕作用。方法:体外实验中,在精液中加入不同浓度的PHA,对使不同物种精子发生凝集作用影响因素进行观察,高浓度PHA制动的精子用相应获能液洗涤后复苏;在体内实验中,对交配见阴栓后的雌鼠于不同时间一次性经阴道给予PHA;另外不同时间间隔对正常性周期小鼠多次给予PHA观察其避孕效果。结果:体外实验结果表明,人、小鼠和牛的精子在低浓度PHA(15.6～125μg/ml)作用下30min就完全发生凝集,但不会致死,不同物种的精子凝集方式有所不同。凝集后的精子可以在室温下(22±7℃)存活7～12h。而≥250μg/ml的高浓度PHA会使这5种精子完全制动并发生不可逆致死。并且在低pH值时,精子存活时间明显缩短。体内实验结果表明经阴道给予PHA具有抗早孕的效果;每天经阴道给予PHA具有100%的避孕效果。结论:在体外实验中,云南大白芸豆植物凝集素具有广泛的精子凝集作用;在体实验证明PHA可有效用于避孕和抗早孕,因此PHA有可能开发成为具有外用避孕作用的药物。     

鹿藿正丁醇提取物体外杀精作用及机制探讨

目的研究鹿藿正丁醇提取物(n-Butanol extract of Rhynchosia volubilis Lour,BERVL)的体外杀精子作用及机制。同时观察该提取物体外对嗜酸乳酸杆菌的影响,初步评估BERVL作为杀精子剂的可行性和安全性。方法体外杀精子实验以生理盐水为阴性对照,壬苯醇醚(nonoxynolum-9,N-9)为阳性对照,显微镜下观察浓度为250mg/ml,125mg/ml,100mg/ml,90mg/ml,62.5mg/ml的BERVL对人精子活力的影响;体外抑菌实验用试管稀释法和琼脂稀释法;体外杀精子机制探讨:用伊红Y水试验和Hoechst 33342/PI荧光双染,显微镜下检测精子膜功能和精子凋亡。结果 BERVL在90mg/ml浓度组:体外杀精实验,原地活动精子百分率为(4.730±3.231)%;体外抑菌试验,对嗜酸乳酸杆菌抑制率为32.96%³5.70%;伊红Y水溶液染色,精子头部未着色百分率为(7.846±4.706)%。Hoechst 33342/PI荧光双染,凋亡精子百分率为(4.977±2.267)%,死精子百分率为(90.111±4.509)%。结论90mg/ml为BERVL较安全理想的体外杀精浓度。BERVL有显著的体外杀精子作用,可能与破坏精子膜和加速精子凋亡及死亡有关。     

波斯迪卫生栓的体内外杀精作用和抗生育作用的实验研究

本文目的是观察波斯迪原料药和卫生栓体内外杀精作用与抗生育作用。体外杀精作用结果表明原料药20sec内最低杀精浓度在人、家兔和大鼠分别为1.25mg/ml,2.5mg/ml和1.25mg/ml。卫生栓则分别为5mg/ml,10mg/ml和5mg/ml。体内杀精作用结果表明0.5g和1g原料药和卫生栓在家兔交配后10min和8min内阴道均未发现有活动精子。家兔抗生育试验结果表明0.25g、0.5g和1g原料药避孕率均为100％(P<0.001)。卫生栓0.25g避孕率为60％(P>0.05),但0.5g和1g均为100％(P<0.001)。本文结果指出由中草药组成的波斯迪卫生栓体内外均有杀精作用和抗生育作用。     

棉酚在猕猴阴道内的杀精子活性

最近报道,在试管内棉酚—聚乙烯吡咯烷酮(gossypol—polyvinylpyrrolidone缩写gossg—pol—pvp)混合沉淀物是一种有效的杀精子剂。使用这种混合沉淀物是因为棉酚不易溶解。gossyp-ol-pvp)浓度为40mg/ml在20秒钟内使精子完全停止运动。而5mg/ml时则需3分钟。目前市场上销售的杀精子剂在试管内起杀精子作用亦需相同浓度。本实验用非人灵长类动物进行体内杀精子活性的观察。     

《维妮好一推得》体外杀精和对家兔的阴道刺激试验

《维妮好一推得》是以卫生棉条做载体,由苦参、蛇床子、土茯苓、川椒、榆皮、土槿皮等中药和壬苯醇醚-9(NP-9)复方配制的药液组成的妇科消毒药剂。本实验采用改良Sander-Cramer方法评价其体外杀精作用。采用雌性实验家兔,分自然对照组、生理盐水对照组、NP-9对照组及《维妮好一推得》高剂量组和低剂量组,阴道给药,进行组织学炎症刺激程度的判定。结果表明《维妮好一推得》的体外杀精作用是单用NP-9的10倍;载体和药液共同留置阴道在试验应用剂量下对阴道无明显刺激作用,评分在可接受范围内。     

Synergistic effect of gramicidin and EDTA in inhibiting sperm motility and cervical mucus penetration in vitro

Gramicidin, a linear polypeptide with antiviral and antimicrobial properties, was compared in vitro with a commonly used spermicidal detergent—nonoxynol-9 (N9). The inhibition of sperm functions was evaluated by computer-assisted semen analysis (CASA) for sperm motility, in cervical mucus penetration assay, and by colorimetric tetrazolium salt and lactate dehydrogenase release assays routinely employed for testing the toxicity of drugs. The effective 100% inhibitory concentration (IC₁₀₀) of gramicidin in a 2-min sperm immobilization assay by CASA was equal to 4 μg/ml, whereas IC₁₀₀ of N9 was equal to 200 μg/ml. The presence of 0.1% of chelating agent, EDTA, reduced IC₁₀₀ of gramicidin to 10 ng/ml, while less than a twofold enhancement in N9 activity was observed upon combination with EDTA. Likewise, the gramicidin/EDTA combination was 100,000 times more potent than N9/EDTA in the sperm penetration assay. Quantitative toxicity tests confirmed that gramicidin is a potent spermostatic rather than spermicidal agent. Further development of a gramicidin/EDTA formulation is warranted as a nontoxic topical contraceptive with activity against viral and microbial sexually transmitted diseases (STDs).     

Sperm immobilizing effect of triterpene saponins from Acacia auriculiformis

A mixture of two partially triterpenoid saponins (Tg), isolated from Acacia auriculiformis was tested for sperm immobilizing activity by using in vitro system. The lowest concentration (ED) required for an obvious immobilization of human sperm by using a modified Sander-Cramer test was found to be 0.35 mg/ml in physiological saline. The ability of the compound as a sperm immobilizing agent was compared with that of Triton X-100 and found to be more potent. Cervical mucus penetration test was also performed and the ED successfully prevented sperm entry in human cervical mucus. Supra-vital staining with eosin-nigrosin indicated death of the treated sperm. Electron microscopic study of Tg-treated sperm showed plasma membrane disintegration and dissolution of acrosomal cap which is presumably the cause for the spermicidal effect of the saponins. No permanent lesion was observed after application of 1.25 mg/ml Tg solution in physiological saline to the eye of rabbits for ten consecutive days.     

Spermicidal activity of dermaseptins

ObjectiveThe present study was undertaken to elucidate the spermicidal efficacy of two synthetic antimicrobial peptides, dermaseptin (DS1 and DS4).MethodsTwenty samples of fresh semen were obtained from patients aged between 23 and 35 years. The ability of DS to kill sperm was evaluated by the Sander–Cramer test under in vitro conditions.ResultsThe data showed that sperm motility was inhibited with various concentrations of DS at different intervals ranging from 2 to 240 min. The effective 100% inhibitory concentration (EC₁₀₀) of DS4 in 2 min during the sperm immobilization assay was 100 μg/mL whereas the sperm immobilization of EC₁₀₀ of DS1 was 200 μg/mL. The presence of 0.1% chelating agent ethylenediaminetetraacetic acid (EDTA) reduced the EC₁₀₀ of DS4 to 10 μg/mL whereas less than a two-time enhancement in DS1 activity was observed upon combination with EDTA. The action of DSs on sperm motility was observed to be dose dependent. Supplementation with pentoxifylline and that with calcium are known to enhance the motility of sperm but they did not prevent the spermicidal action of DSs.ConclusionThis present study indicates that DS is an effective agent to kill sperm. In view of this fact, it is suggested that DS4 has antibacterial, antiviral, antifungal and potentially spermicidal activities and could be a potent vaginal contraceptive.     

Potent spermicidal effect of oleanolic acid 3-beta-D-glucuronide, an active principle isolated from the plant Sesbania sesban Merrill

Background

The spermicidal activity of oleanolic acid 3-β-d-glucuronide (OAG), an active principle isolated from root extracts of Sesbania sesban, was evaluated.

Study Design

Under the Sander–Cramer test criteria, the sperm-immobilizing activity of OAG was studied using highly motile rat sperm. Sperm mortality and membrane integrity were assessed by supravital staining, hypo-osmotic swelling (HOS), transmission electron microscopy (TEM) and sperm membrane lipid peroxidation (LPO). In vitro microbicidal potential and hemolytic index of OAG were examined in Lactobacillus culture and rat red blood corpuscles (RBCs), respectively. Post-intravaginal OAG application, the in vivo contraceptive efficacy was evaluated in rats. Ames test determined the carcinogenic potential of OAG.

Results

The minimum effective concentration (MEC) of OAG was 50 mcg/mL. More than 97% of the OAG-treated sperm lost their HOS responsiveness in a dose-dependent manner. TEM and LPO revealed that OAG affected the sperm membrane integrity. OAG declined fertility to zero, was nonmutagenic and was not harmful to lactobacillus.

Conclusion

OAG has significant spermicidal activity that may be explored further.     

Spermicidal effects of amphotericin B and nystatin on bull and rabbit sperm and contraceptive effects in rabbits

Vaginal contraceptives have potential for controlling reproduction as well as sexually transmitted diseases. Two extensively used fungicides, amphotericin B and nystatin, were found to be highly spermicidal to bull and rabbit sperm even in the presence of organic material found in milk and egg yolk, both excellent components of media designed to preserve sperm. In repeated tests with many samples of bull and rabbit sperm, as little as 0.5 micro g/mL of amphotericin B completely immobilized sperm after prolonged exposure, and 1000 micro g/mL immobiled sperm in less than 1 min. Treatment of rabbit sperm with 1000 micro g of solubilized amphotericin B in 0.4 ml of a glucose-semen mixture before insemination resulted in a total of 2 young born from 9 inseminated Dutch does, compared with 55 young from 9 control does. As this inhibition was achieved without the use of enhancing foams or gels, these results provide promising leads for contraceptive research.     

中药远志提取物体外杀精子效果与机制研究

目的:通过精子低渗肿胀(HOS)实验结合改良伊红染色法观察中草药远志粗提物体外杀精活性及机制。方法:HOS实验结合伊红-姬姆萨(EG)染色(HOS-EG)判断精子膜完整性,将精子分为A(HOS+-EG-)、B(HOS-EG-)、C(HOS+-EG+)、D(HOS-EG+)4型。精子染色质扩散实验(SCD)检测精子DNA碎片。制备远志粗提物,并判断其体外杀精子活性。壬苯醇醚-9(N-9)作为标准参照物,生理盐水为对照组。结果:远志粗提物浓度为50.0mg/ml和20.0mg/ml时体外20s内即可完全杀灭精子(与精液1:1混合)。对照组、远志粗提物组及N-9组精子DNA碎片比例各组比较无差异。对照组A、B、C和D型精子百分比分别为(69.0±8.3)%、(3.4±0.5)%、(10.2±1.7)%和(17.4±2.1)%。远志粗提物组及N-9组精子膜完整性全部被破坏,无A型和B型精子,C型精子较对照组明显减少,D型精子比对照组明显增加(P<0.001)。精子复活实验显示远志粗提物组和N-9组均无活动精子恢复。结论:HOS-EG实验可以同时判断一条精子头膜或尾膜损伤。远志粗提物在体外通过破坏精子膜使精子快速失活。     

Attainment and maintenance of azoospermia with combined monthly injections of depot medroxyprogesterone acetate and testosterone enanthate

Depot medroxyprogesterone acetate (DMPA) and testosterone enanthate (TE) were administered as a monthly treatment to 20 normal male volunteers. Two regimens were employed: 1) 1000 mg DMPA and 250 mg TE initially and repeated after 1 month, followed by 150 mg DMPA and 250 mg TE at each sucessive month; 2) 1000 mg DMPA and 250 mg TE initially, followed by 300 mg DMPA and 250 mg TE at each successive month. Sperm production was severely inhibited in the 19 subjects who were followed beyond 2 months: azoospermia in 14 subjects; less than 1 million sperm/ml in 2 subjects; 1–2 million sperm/ml in 2 subjects; and 3 million sperm/ml in 1 subject. The lowest sperm count attained (azoospermia or severe oligozoospermia) was usually maintained for at least several months; then, despite continued treatment, most subjects eventually showed at least slight increases in sperm concentration. However, since only 1 sperm count rose above 20 million/ml, and since sperm motility remained poor, the subjects may actually have been infertile. Gonadotropins and testosterone decreased initially, then tended to recover somewhat in later months of treatment. The only side effects observed were moderate weight gain and some slight increases in serum transaminases. There were no changes in other blood values, in blood pressure, in the size, consistency or sensitivity of testicles, prostate or breasts, or in libido/sexual behavior. The few differences in the effects of the 2 different regimens were not striking.