Assessment of reversible contraceptive efficacy of methanol extract of Mentha arvensis L. leaves in male albino mice

The present study was undertaken to assess the reversible contraceptive efficacy of methanolic extract of Mentha arvensis leaves. Aqueous solution of the extract (10 mg per day per mouse) when administered orally to male mice of proven fertility for 20, 40 and 60 days caused inhibition of fertility while maintaining their normal sexual behaviour. With the increase in treatment duration, there occurred a corresponding decrease in the mean weight of testis and accessory organs of reproduction. Sperm concentration, motility and viability in the cauda epididymis were also decreased. Spermatozoa with coiled tails also appeared in the epididymal smear. However, all the induced effects returned to normalcy within 30 days following withdrawal of 60-day treatment. Oral administration of the extract also did not affect the body weight of the mice and their blood cells count, packed cell volume, haemoglobin and blood/serum biochemistry.     

Post-testicular antifertility effects of Abrus precatorius seed extract in albino rats

Oral administration of a 50% ethanol extract of Abrus precatorius seeds (250 mg/kg) in albino rats for 30 and 60 days induced an absolute infertility in males which was reversible. Suppression of sperm motility in the cauda epididymis was the most pronounced effect of the treatment. Such treatment may affect the oxidative/energy metabolism of the cauda epididymis. Histological and histocytometric observations in testis and parareproductive tissues appeared normal while the protein, sialic acid, acid phosphatase and succinic dehydrogenase levels were significantly depleted.     

Antifertility effect of chronically administered Martynia annua root extract on male rats

The effects of 50% ethanol extract of Martynia annua L. root on reproduction was studied on male rats. The study was divided into four groups of five animals each. The first group (I) received vehicle alone to serve as control. The second, third and fourth groups (II, III and IV) of animals were administered the root extract daily at 50 mg/kg body weight, po, 100 mg/kg body weight, po, and 200 mg/kg body weight, po, respectively, for a period of 60 days. Significant decreases in the weights of testes, epididymides, seminal vesicle and ventral prostate were observed. A dose related reduction in the testicular sperm count, epididymal sperm count and motility, number of fertile males, ratio between delivered and inseminated females and number of pups were observed. The testis showed a clear correlation between the dose and severity of lesions of seminiferous epithelium. In general, the seminiferous tubules appear reduced in size with a frequently filled eosinophilic material. Spermatogenesis arrested at the secondary spermatocyte stage. Pachytene spermatocytes were undergoing degeneration. Disorganisation and sloughing of immature germ cells were visible. Leydig cells were atrophied. No morphological changes were observed in Sertoli cells. Significant reduction in serum concentration of luteinising hormone and testosterone were observed. No distinct change in serum FSII concentration was recorded. The final body weights of all groups were elevated markedly. No alterations were recorded in any hematological parameters. It is concluded that the 50% ethanol extract of M. annua root produced dose related effects on male reproduction without altering general body metabolism.     

Effects of bamboo buds: structural and functional changes in the epididymis of rats

An ethanolic extract of the tender shoots of Bambusa arundinaceae was administered at 300 mg/kg per rat per day for 7 days to adult male rats to assess epididymal structural and functional activity. Sperm motility decreased markedly in the cauda epididymal fluid and sperm count decreased significantly in both caput and caudal segments of the epididymis. Histologically, a reduction in epithelial and stereocilia height (in both segments) and lumen diameter (in cauda) was noted. An increase in intertubular stroma was also evident. Epididymal weights, activities of acid phosphatase and total LDH were reduced in both epididymal segments. Protein concentration was appreciably increased only in the caudal segment. Extract therapy impaired the structural and functional integrity of the epididymis.     

Ultrastructural changes in the testis and epididymis of rats following treatment with the benzene chromatographic fraction of the chloroform extract of the seeds of Carica papaya

The benzene chromatographic fraction of the chloroform extract of the seeds of Carica papaya at a dose of 10 mg/rat/day for 150 days, which has shown a total inhibition of motility, reduced sperm count and infertility, was tested to de fi ne the mode of action at the subcellular level in the testis and epididymis. The ultrastructure of the testis of the treated animals revealed no appreciable changes in the subcellular characteristics. The mechanism of protein synthesis as well as steroidogenesis were evident in the Sertoli cells while the spermatogonia, spermatocytes and spermatids, both round and elongated, depicted a prominent nucleus, distinct nuclear membrane and cytoplasmic characteristics indicating normal germ cell differentiation. The principal cells of the cauda epididymis were characterized by the presence of well-de fi ned rough endoplasmic reticulum, mitochondria, Golgi bodies and secretary granules, suggesting active secretory functions. The absorptive function of the cauda epididymis was evidenced by the presence of numerous vesicles and multivesicular bodies adjacent to stereocilia. It is concluded that the inhibition of sperm motility by the drug could be due to other epididymal factors rather than the subcellular characteristics of testis and epididymis.     

Antifertility investigation and toxicological screening of the petroleum ether extract of the leaves of Mentha arvensis L. in male albino mice

In male albino mice, the petroleum ether extract of the leaves of Mentha arvensis L., at the doses 10 and 20 mg/mouse per day for 20, 40 and 60 days, when administered orally, showed a dose and duration dependent reduction in the number of offspring of the treated male mated with normal females. Negative fertility was observed in both dose regimens after 60 days of treatment. The body weight and libido of the treated ammals remain unaffected. However, a significant decrease in the weight of the testis, epididymis, cauda epididymal sperm count, motility, viability and normal morphology of the spermatozoa was observed. The levels of serum protein, bilirubin, GOT, GPT and acid phosphatase, blood urea and haematological indices were unaltered throughout the course of investigation. All the altered parameters were reversible following withdrawal of treatment. The results suggest that the petroleum ether extract of the leaves of M. arvensis possess reversible antifertility property without adverse toxicity in male mice.     

鹿藿醇提取物的体外杀精及抗菌作用

目的研究鹿藿醇提取物的体外杀精及抗菌作用。方法(1)鹿藿醇提取物按照国际计划生育基金会认可的杀精子实验方法,对10只小鼠附睾精子悬液和10例人精液进行体外杀精实验。(2)用试管法及琼脂平皿稀释法观察鹿藿醇提取物对几种临床常见致病菌的体外抑菌作用。结果鹿藿醇提取物对小鼠和人精子最低杀精浓度分别为50和100mg/ml;对标准菌金黄色葡萄球菌、铜绿假单胞菌、大肠埃希菌、肺炎克雷伯菌MIC(试管法)分别为1.0,1.0,4.0,4.0 mg/ml。对临床分离菌金黄色葡萄球菌、铜绿假单胞菌、大肠埃希菌MIC(试管法)分别为0.5,2.0,8.0 mg/ml。结论鹿藿醇提取物对小鼠和人精子有一定杀精作用,对临床常见致病菌有比较强的抗菌作用。     

Androgenic and Spermatogenic Activity of Alkylamide‐Rich Ethanol Solution Extract of Anacyclus pyrethrum DC

Anacyclus pyrethrum (A. pyrethrum) has been used as Vajikaran Rasayana (aphrodisiac) in traditional Indian ayurvedic medicine to treat male sexual dysfunction, including infertility. Aphrodisiac activity may be due to an increase in the production or effect of androgens, so this study sought to evaluate the androgenic and spermatogenic potential of the alkylamide‐rich ethanol solution extract. Male Wistar strain rats weighing between 150 and 180 g were completely randomized divided into five groups. The ethanol solution extract of A. pyrethrum was administered to groups of rats in 50, 100, and 150 mg/kg doses for a period of 28 days, and the action was compared with control and testosterone‐treated rats. Thirteen N‐alkylamides were detected in the extract by using HPLC/UV/electrospray ionization mass spectrometry method. Extract administration at all the doses produced significant increase in body weight, sperm count, motility, and viability along with serum testosterone, luteinizing hormone, and follicle‐stimulating hormone concentrations. Histoarchitecture of testis revealed increased spermatogenic activities. Seminal fructose content was also significantly increased after 28 days of treatment. Our results suggest that the ethanol solution extract of the roots of A. pyrethrum has androgenic potential and may improve male fertility by enhancing spermatogenesis. Copyright © 2012 John Wiley & Sons, Ltd.     

The effect of the aqueous extract of Cynomorium coccineum on the epididymal sperm pattern of the rat

An aqueous extract of Cynomorium coccineum was administered by stomach tube to ten mature male Wistar rats, at a dose of 47 mg/100 kg body weight/day for 14 consecutive days. Ten rats were kept as controls and received normal saline by oral route at the same dosing interval. Sperm was collected from the epididymes after decapitation. The results revealed that the water extract of the Cynomorium coccineum induced significant increase in the sperm count, improved the percentage of live sperm and their motility and decreased the number of abnormal sperm. Testicular histology showed increased spermatogenesis and seminiferous tubules full of sperm in the treated group compared with the controls. Copyright © 1999 John Wiley & Sons, Ltd.     

Genistein Mitigates Radiation-induced Testicular Injury

The present study investigated the radioprotective effect of a multifunctional soy isoflavone, genistein, with the testicular system. Genistein was administered (200 mg/kg body weight) to male C3H/HeN mice by subcutaneous injection 24 h prior to pelvic irradiation (5 Gy). Histopathological parameters were evaluated 12 h and 21 days post‐irradiation. Genistein protected the germ cells from radiation‐induced apoptosis (p < 0.05 vs vehicle‐treated irradiated mice at 12 h post‐irradiation). Genistein significantly attenuated radiation‐induced reduction in testis weight, seminiferous tubular diameter, seminiferous epithelial depth and sperm head count in the testes (p < 0.05 vs vehicle‐treated irradiated mice at 21 days post‐irradiation). Repopulation and stem cell survival indices of the seminiferous tubules were increased in the genistein‐treated group compared with the vehicle‐treated irradiation group at 21 days post‐irradiation (p < 0.01). The irradiation‐mediated decrease in the sperm count and sperm mobility in the epididymis was counteracted by genistein (p < 0.01), but no effect on the frequency of abnormal sperm was evident. Reactive oxygen species (ROS) were evaluated using DCFDA method and exposure to irradiation elevated ROS levels in the testis and genistein treatment resulted in a significant attenuation of radiation‐induced ROS production. The results indicate that genistein protects from testicular dysfunction induced by gamma‐irradiation by an antiapoptotic effect and recovery of spermatogenesis. Copyright © 2011 John Wiley & Sons, Ltd.     

Contraceptive efficacy of Carica papaya seed extract in male mice (Mus musculus)

The effects of an aqueous extract of Carica papaya seeds (20 mg/kg body weight/animal/day orally and 5 mg/kg body weight/animal/day i.m. for 60 days) were investigated for contraceptive efficacy and other related side effects in male albino mice, Mus musculus. The data revealed that the extract might be causing an androgen deprived effect to target organs resulting in alterations in the internal milieu of the cauda epididymis especially. The treatment did not, however, affect the testicular sperm count suggesting that it acted at the post-testicular level which lead to a reduction of cauda epididymal sperm motility and thus the treatment brought about a significant reduction in fertility rate. The induced effects were transient and reversible upon withdrawal of the treatment, elucidating that functional sterility could be induced by the aqueous papaya seed extract in rodents.     

杜仲补天素胶囊对白消安诱导的生精障碍小鼠的影响

目的研究杜仲补天素胶囊(Duzhong Butiansu Capsule,DBC)对白消安致生精功能障碍的影响。方法Balb/c雄性小鼠腹腔注射白消安制备生精障碍模型。造模成功后,生精组给予0.8 g·kg^-1生精胶囊,褪黑素组给予10 mg·kg^-1褪黑素,杜仲补天素低剂量组和杜仲补天素高剂量组分别给予0.694 g·kg^-1、1.388 g·kg^-1杜仲补天素,共给药35 d。称量脏器,用精子分析仪检测精子活力及数目;采用流式细胞仪检测睾丸细胞周期;进行HE染色,观察睾丸、附睾和肝的形态学结构。同时,检测了血清促卵泡生成素(Follicle stimulating hormone, FSH)、促黄体生成素(Luteinizing hormone, LH)、睾酮(testosterone, T)、血清丙二醛(malondialdehyde,MDA)、血清超氧化物歧化酶(superoxide dismutase,SOD)、肝中谷胱甘肽(glutathione,GSH)水平。采用免疫组织化学法测定了睾丸c-kit、Caspase-3、Cleaved Caspase-3的表达水平。结果与正常组比较,白消安模型组明显降低了精子活力及数目以及睾丸细胞单倍体、二倍体的比例(P<0.01),且破坏睾丸、附睾和肝的组织形态;显著降低血清T(P<0.01)水平,升高FSH、LH和氧化应激水平(P<0.05,P<0.01);明显降低c-kit的表达水平(P<0.05),并明显升高Caspase-3和Cleaved Caspase-3的表达(P<0.01)。与模型组比较,经DBC作用后,精子活力及二倍体、四倍体比例升高(P<0.05,P<0.01),同时保护睾丸、附睾和肝;改善血清性激素水平及氧化应激水平;升高c-kit表达水平(P<0.01),降低凋亡蛋白Caspase-3、Cleaved Caspase-3的表达水平(P<0.05,P<0.01)。结论杜仲补天素胶囊可改善白消安诱导的生精障碍,效果以低剂量为佳。     

Antifertility effects of beta-sitosterol in male albino rats.

The effects of beta-sitosterol on fertility, epididymal sperm counts and testicular and accessory reproductive organ weights were evaluated in male albino rats. The effects were studied at two dosages (0.5 and 5 mg/kg per day rat subcutaneously) for 16, 32 and 48 days. The antifertility effect of beta-sitosterol was pronounced only at the high dose level, but there was a significant decrease in testicular weight and sperm concentrations after long-term treatment with the low dose of beta-sitosterol. The weights of all accessory sex tissues except caput epididymis increased following low dose sitosterol treatment. High dose treatment reduced the sperm concentrations as well as the weights of testis and accessory sex tissues in a time-dependent manner. Withdrawal of treatment for 30 days restored only the weights of accessory sex tissues to near normal conditions.     

Effects of Ginsenoside Rb2 and Rc on inferior human sperm motility in vitro

The purpose of the study was to investigate the effects of two constituents of Panax notoginseng flower extract, Ginsenoside Rb2 and Rc, on human sperm motility and progression in vitro. Semen samples were collected from 20 patients with sperm motility between 20% and 40% of normal. All samples had sperm counts of over 20 million per milliliter, in accordance with the World Health Organization standard. Sperm were separated by a Percoll discontinuous gradient technique, and divided into a Percoll sperm control group, and three Ginsenoside Rb2 experimental groups (0.1, 0.01 and 0.001 mg/ml) and three Ginsenoside Rc experimental groups (0.1, 0.01 and 0.001 mg/ml). The results showed that at concentrations of 0.01 mg/ml and 0.001 mg/ml, Ginsenoside Rc enhanced both sperm motility and sperm progression significantly at the end of the 1st and 2nd hour. However, the three concentrations of Ginsenoside Rb2 did not increase sperm motility at the 1st or 2nd hour, but promoted sperm progression at the 2nd hour, when compared to the Percoll group.     

Sperm antimotility properties of a seed extract of Abrus precatorius

This study examined the inhibitory effects of a methanol extract of Abrus precatorius seeds on the motility of washed human spermatozoa. The extract caused a concentration-related impairment of percentage sperm motility; with the EC50 concentration being 2.29 mg/ml. This effect on motility was essentially irreversible. With the highest concentration tested (20.0 mg/ml), the onset of the antimotility action was almost immediate. In addition, this concentration impaired the functional integrity of the plasma membrane (hypoosmotic swelling test) and viability (nigrosin-eosin stain) of spermatozoa. In contrast, with a lower concentration (5.0 mg/ml), such effects were not evident. It is concluded that at the lower concentrations the antimotility action may result from a rise in intracellular calcium (not via influx) and/or a decline in cAMP content and/or enhanced generation of a reactive oxygen species.     

Water extracts from Momordica charantia increase glucose uptake and adiponectin secretion in 3T3-L1 adipose cells

To examine the effects of Momordica charantia on glucose uptake and adiponectin secretion in adipose cells, 3T3-L1 adipocytes were treated with three concentrations (0.2, 0.3 and 0.4mg/ml) of water and ethanol extracts of Momordica charantia fruit and seeds alone and in combination with either 0.5nM or 50nM insulin. The treatment combination of 0.2mg/ml water extract and 0.5nM insulin was associated with significant (p<0.05) increases in glucose uptake (61%) and adiponectin secretion (75%) over control levels. The ethanol extract was not associated with an increase in glucose uptake; however, a dose-dependent decrease in basal glucose uptake and insulin-mediated glucose uptake was observed with the ethanol extract in combination with 50nM insulin. In the absence of insulin, no effects on glucose uptake were observed in adipocytes exposed to the water extracts whereas the highest concentration (0.4mg/ml) of the ethanol extract was associated with a significant (p<0.05) decrease in glucose uptake relative to controls. The present results indicate that water-soluble component(s) in Momordica charantia enhance the glucose uptake at sub-optimal concentrations of insulin in 3T3-L1 adipocytes, which is accompanied by and may be a result of increased adiponectin secretion from the 3T3-L1 adipocytes.     

Beneficial effects of quercetin on sperm parameters in streptozotocin‐induced diabetic male rats

Quercetin (QR) is a strong antioxidant and has been shown to reduce oxidative stress in the long‐term treatment of streptozotocin (STZ)‐induced diabetes in animal models. Antioxidants have significant effects on spermatogenesis, sperm biology and oxidative stress, and changes in antioxidant capacity are considered to be involved in the pathogenesis of chronic diabetes mellitus. The present study aims to examine the influence of QR on spermatogenesis in STZ‐induced diabetes in male Wistar rats. Animals (n = 50) were allocated into five groups: Group 1: Control rats given 0.5 ml of 20% glycerol in 0.9% normal saline. Group 2: Control rats given buffer (pH4.0).Group 3: diabetic controls. Group 4: rats given QR 15 mg/kg/day (i.p.). Group 5: STZ + QR rats. Animals were kept in standard conditions. At the end of the experiment (28th day), blood samples were taken for determination of testosterone, total antioxidant capacity, and levels of malondialdehyde and oxidized low‐density lipoprotein. All rats were euthanized, testes were dissected out and spermatozoa were collected from the epididymis for analysis. Sperm numbers, percentages of sperm viability and motility, and total serum testosterone increased significantly in QR‐treated diabetic rats (P < 0.05) compared with control groups. In histopathology, degeneration and inflammation in testes cells associated with diabetes were improved and testes weights in the QR‐treated diabetic group decreased significantly in comparison with controls (P < 0.05). We conclude that QR has significant beneficial effects on the sperm viability, motility, and serum total testosterone and could be effective for maintaining healthy sperm parameters and male reproductive function in diabetic rats. Copyright © 2010 John Wiley & Sons, Ltd.     

鹿藿醋酸乙酯提取物抗雄性小鼠生育研究及成分分析

目的：研究鹿藿醋酸乙酯提取物(Rhynchosia volubilis Lour ethyl acetate extract,RVLEAE)的抗生育作用及化学成分分析。方法：①用溶剂萃取法提取有效部位,减压蒸馏浓缩制成浸膏；②抗生育实验：80只雄性小鼠分为4组即生理盐水(NS)对照组,阳性药物(雷公藤多苷10 mg·kg^-1·d^-1)对照组,鹿藿醋酸乙酯提取物低、高剂量(醋酸乙酯浸膏100,400 mg·kg^-1·d^-1)组,灌胃,每天1次,连续给药11周。给药2,10周后,合笼交配,1周后处死部分雄鼠观察附睾精子、附睾及睾丸病理变化、血清睾酮含量以及体重变化情况；分笼10 d后处死雌鼠,观察妊娠率、活胎数、死胎数；③化学成分分析用柱层析分离、薄层色谱初步分析及红外色谱、核磁共振波谱定性分析。结果：RVLEAE连续用药2周,小鼠妊娠率明显下降,妊娠抑制率明显增高。连续用药10周后作用仍较明显。3周时的精子数量,在高、低剂量组减少最为明显。连续给药后精子数量减少。RVLEAE对小鼠睾丸及附睾组织影响不大。血清睾酮含量,阳性对照组明显增高,而高、低剂量组与生理盐水对照组差异无显著性。RVLEAE的主要成分为糖及苷类、醇和酚类化合物。3号组分被鉴定为β-甲基2,4-二甲氧基-L-吡喃岩藻糖苷。6号组分被鉴定为α,β-D-吡喃葡萄糖和α,β-甲氧基-D-吡喃葡萄糖单糖的混合物。结论：RVLEAE有抗生育作用,作用部位主要在附睾,作用机制可能是糖苷类化合物干扰了精子在附睾的成熟。     

Evaluation of contraceptive activity of methanol extract of Dendrophthoe falcata stem in male albino rats

The present study was undertaken to evaluate the contraceptive efficacy of methanol extract of Dendrophthoe falcata Ettingsh (family-Loranthaceae), stem in male albino rats as reported in folk remedies. Adult proven fertile male rats were gavaged methanol extract of D. falcata stem at 50, 100 and 200mg/rat/day for 60 days. The activity was compared with standard drug, i.e. Lonidamine. On day 61 the animals were autopsied and the testes, epididymides, seminal vesicle and ventral prostate were dissected out and weighed. Sperm motility and density and serum testosterone level were assessed. The sperm motility and density were significantly reduced. The histoarchitecture of testes revealed degenerative changes in the seminiferous tubules, arrest of spermatogenesis at the stage of round spermatid. Serum testosterone levels were decreased significantly in all treatment groups. It is concluded that D. falcata methanol stem extract showed a significant effect on fertility in male rats as reported in folk remedies.