Evaluation of Andrographis paniculata-supplemented Diet on the reproductive indices of Plasmodium berghei-infected mice.

ETHNOPHARMACOLOGICAL RELEVANCE: The King of Bitters (Andrographis paniculata) is a plant used to cure a wide range of infectious diseases which includes malaria, fever and others. However, there is a paucity of scientific evidence of its effect on male reproductive indices during malaria treatment. AIM OF THE STUDY: The aim of this study is to evaluate the effect of supplemented diet on antiplasmodial, hematological and male reproductive indices in mice infected with Plasmodium berghei. MATERIALS AND METHODS: Aqueous extract of A. paniculata (King of Bitters, KGB) was prepared and the total phenol and flavonoid contents were determined. Forty-two mice, weighing 20-25 g, were distributed into 7 groups consisting of 6 mice each. The mice were innoculated with strain NK65 Plasmodium berghei (Chloroquine, CQ sensitive) and the parasitemia suppression was assessed. The mice were fed with the dietary supplementation of KGB at varying inclusions (2.5%, 5%, 7.5%, and 10%) and administered 10 mg/kg CQ (which served as the positive control) for 5 consecutive days after infection was established. The reactive malondialdeahyde (MDA), antioxidant [superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH)] and the hematological (hemoglobin, packed cell volume and red blood cell) parameters in the infected mice were determined. The reproductive indices (serum testosterone, luteinizing hormone (LH), follicle-stimulating hormone (FSH), sperm count, sperm motility, and sperm viability) and testis histopathology were also assessed. RESULT: The result revealed that KGB had a total phenol content of 32.55 mgGAE/g and total flavonoid content of 19.71 mgQUE/g. The infected mice treated with the dietary supplementation of KGB showed significantly decreased (p < 0.05) parasitaemia and MDA levels. Furthermore, the 7.5% dietary inclusion showed significant improvement in the antioxidant, hematological and reproductive indices as well as the restoration of testis morphology as seen in the histopathology plate of the infected mice treated with KGB. Hence, this study suggests that the KGB- supplemented diet (7.5%) may be a potential alternative and complementary therapy in the treatment of malaria infection and reproductive disorders.

Effect of Medicago sativa seed powder (Plus vitamin E vs. vitamin E alone) on semen analysis in men with idiopathic infertility: A double blind randomized clinical trial.

ETHNOPHARMACOLOGICAL RELEVANCE: Different parts of the alfalfa plant (Medicago sativa L.), especially its seeds, have been introduced as a semen quality enhancer in the folk medicine of different regions of Iran as well as in the traditional Persian medicine (PM) books. The seeds of this plant are also used in many combined medicines to treat male infertility in PM. AIM OF THE STUDY: This study was designed to investigate the effect of M. sativa seed powder plus vitamin E vs. vitamin E alone on semen analysis of men with idiopathic infertility. MATERIALS AND METHODS: In this randomized clinical trial, 60 participants were randomly divided into two groups and then received M. sativa seed powder (6 g daily) or placebo for three months, as add-on to vitamin E (100 U). Results of semen analysis (sperm total count, motility, and morphology as well as semen volume) of these two groups and their changes were compared before and after treatment as the primary outcome. The number of pregnancies after one month and probable side effects were also assessed. RESULTS: After 12 weeks, sperm total count changed from 36.2 ± 21.8 × 106/ml to 48.5 ± 19.1 × 106/ml in M. sativa and from 39.5 ± 23.5 × 106/ml to 41.2 ± 20.9 × 106/ml in placebo, percentage of normal morphology changed from 1.8 ± 0.8% to 2.6 ± 1.2 % in M. sativa and from 2.0 ± 0.9% to 2.6 ± 1.2% in placebo and percentage of motile sperm changed from 36.5 ± 11.8 % to 39.7 ± 12.0% in M. sativa and from 39.3 ± 10.1 % to 38.1 ± 12.1% in placebo. The improvements in M. sativa group are significantly better than placebo group (P = 0.00, 0.01 and 0.04, respectively). However, semen volume decreased in both groups, but its changes were not significant. The number of pregnancies was four in the intervention and zero in placebo group. One case with abdominal bloating and one with respiratory allergy withdrew from the intervention group. No other adverse effect was reported. CONCLUSIONS: In this study, significant improvement was detected in sperm parameters (except semen volume) of men with idiopathic infertility in M. sativa plus vitamin E group in compare to vitamin E alone after 12 weeks. These findings suggest that the M. sativa, as a complementary therapy, may have a beneficial effect on semen quality. More clinical trials with larger sample size are needed.

Beneficial Effect of Methanolic Extract of Frankincense (Boswellia Sacra) on Testis Mediated through Suppression of Oxidative Stress and Apoptosis.

Boswellia sacra oleo gum resin (Burseraceae) commonly known as frankincense is traditionally used in many countries for its beneficial effect on male fertility. This study explores its effect on the male reproductive system after a 60-day repeated administration at two different doses to rats (in vivo) and on human Leydig cells (in vitro). The methanolic extract of B. sacra was analyzed for the presence of various constituents by preliminary phytochemical analysis and gas chromatography-mass spectrometry (GC-MS) while quantitative analysis of boswellic acids was done by high-performance liquid chromatography (HPLC). Administration of B. sacra extract to rats elevated the serum testosterone levels with an associated reduction in serum levels of FSH and LH. An increase in the activity of antioxidant enzymes, superoxide dismutase and catalase, was seen. A dose-dependent increase in the sperm count and sperm motility was also observed. The in vivo results were supported by changes in the expression of the Bcl-2 gene and caspase-3 gene in human Leydig cells in vitro. The results of this study support the traditional use of B. sacra to increase male fertility.

Effects of antioxidant co-supplementation therapy on spermatogenesis dysfunction in relation to the basal oxidation-reduction potential levels in spermatozoa: A pilot study.

Purpose: In this pilot study, the authors compared the effects of antioxidant co-supplementation therapy and methylcobalamin therapy in patients with impaired semen quality. Methods: Eighty-four subjects who visited male infertility clinics and showed abnormal semen test results were randomly subjected to one of the two therapies: antioxidant co-supplementation therapy with vitamin C, vitamin E, coenzyme Q10, and flaxseed oil or methylcobalamin therapy. The oxidation-reduction potential (ORP) and 8-hydroxy-2'-deoxyguanosine levels were used as indicators of oxidative stress levels in semen. Semen analysis was also performed. Results: The authors obtained results from 67 patients who had completed 3 months of treatment. Neither antioxidant co-supplementation therapy nor methylcobalamin therapy changed the semen parameters significantly (except for the sperm concentration, which was increased by the latter therapy). When the pre-treatment ORP value in semen was higher than the cutoff value, both therapies significantly increased the sperm concentration. The 8-hydroxy-2'-deoxyguanosine level did not yield any meaningful predictive value with regard to increased sperm concentrations. Conclusions: Both antioxidant co-supplementation therapy and methylcobalamin therapy increased the sperm concentration in patients with impaired semen quality when the basal ORP levels in their semen were elevated.

Ficus natalensis extract alleviates Cadmium chloride-induced testicular disruptions in albino rats.

BACKGROUND: Cadmium is a potential environmental pollutant with worldwide health problems. Many Ficus species are reported to have an extensive diversity of traditional uses, among them the treatment of reproductive toxicity. OBJECTIVES: This study set out to evaluate the effect of Ficus natalensis extract on the testicular impairments induced by cadmium chloride (CdCl2) and investigated the potential mechanisms associated with its treatment. METHODS: Thus, 40 male albino rats were categorized into 4 groups (n = 10); group I (control), group II (cadmium-treated group) orally received 5 mg/kg/day CdCl2 for one month, group III (cadmium + Ficus natalensis extract) orally received 5 mg/kg/day CdCl2 for one month plus 200 mg/kg/day Ficus natalensis extract for another month, and group IV (cadmium + reference drug (mesterolone) orally received 5 mg/kg/day CdCl2 for one month plus 4.16 mg/kg/day mesterolone for another month. RESULTS: At the end of experiment, CdCl2 administration markedly induced histological and histo-morphometric changes with a substantial (p < 0.05) decrease in the sperm count, sperm motility, serum TAC, serum testosterone, downregulation in the mRNA expression levels of testicular 17ß-HSD and StAR, in addition to a significant increase in serum TNF-α and testicular MDA level compared to the control group. Conversely, the treatment with Ficus natalensis methanolic extract as well as the reference drug significantly ameliorated the above-mentioned adverse effects induced by CdCl2. CONCLUSIONS: Our results suggested that Ficus natalensis extract can attenuate the CdCl2-induced testicular impairments via inhibiting the oxidative cell damage and inflammation that contributed to CdCl2 toxicity.

Duration-dependent effects of high dose of phthalate exposure on semen quality in adult male rats.

OBJECTIVE: To determine the length of exposure to high doses of phthalate that might affect sperm quality in adult male Wistar rats. METHODS: Forty-two (42) adult male Wistar rats (weighing 150-200 g) were randomly assigned into six groups (n=7): Group A received 0.5 mL of distilled water - placebo - and served as controls; groups B, C, D, E and F received Phthalate (750 mg/kgbw) for 1, 3, 5, 7 and 9 weeks, respectively. The data obtained from the study was expressed as Mean ± SEM with a p-value <0.05 considered significant. The data was analyzed with one-way analysis of variance (ANOVA) followed by Tukey's post-hoc test using GraphPad Prism, version 8. RESULTS: The results showed a statistically significant (p<0.05) decrease in testicular weight in the rats exposed to 750 mg/kg of phthalate for 3, 5, 7 and 9 weeks when compared with the controls. Sperm count, motility and viability were also significantly (p<0.05) reduced, while sperm cells with abnormal morphology had increased counts in the groups exposed for 3, 5, 7 and 9 weeks when compared with controls. Serum zinc and magnesium were also significantly reduced (p<0.05) in the subjects treated for 1, 3, 5, 7 and 9 weeks when compared with controls. CONCLUSIONS: The dosage of phthalate adopted in this study was deleterious to testicular function when rats were exposed to it for as short a period as three weeks.

Effect of Opuntia ficus indica extract on methotrexate-induced testicular injury: a biochemical, docking and histological study.

Methotrexate (MTX) is a chemotherapeutic medicine used in the treatment of several types of cancer and inflammatory diseases. It exhibits several drawbacks especially on highly dividing and developing cells. This study aimed to assess the role of Opuntia ficus indica ethanolic extract on testicular damage induced by MTX in rat. MTX was administrated for 10 days (20 mg/kg). Extract of cactus cladodes (Opuntia ficus indica) was given to MTX-treated rats (0.4 g/kg). Spermatozoa were collected from cauda epididymis and analyzed for sperm count and motility. Testis samples were used for histopathological and oxidative stress studies (assessment of malondialdehyde (MDA) levels, protein carbonyls (PCs), catalase (CAT) glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities). Moreover, levels of testosterone were measured in serum by radioimmunoassay. Our results showed that MTX had destructive effects on sperm count and motility associated with significant decrease in testosterone levels in MTX group. This effect was then confirmed by docking results. Testis of MTX group showed increased oxidative stress status. In fact, PCs and MDA were increased and CAT, GPx and SOD were decreased suggesting increased reactive oxygen species and deficiency in enzymatic antioxidant. These findings were associated with disrupted testicular morphology as assessed by histological study. Cladodes extract had protective effects on rat's gonad histology, oxidative stress and improve both sperm parameters (count and motility) and serum testosterone levels. In conclusion, our results suggested that Opuntia ficus indica cladodes extract improved MTX-induced testicular injury and possess potent fertility boosting effects in rats.Communicated by Ramaswamy H. Sarma.

Verbascoside attenuates experimental varicocele-induced damage to testes and sperm levels through up-regulation of the hypothalamus-pituitary-gonadal (HPG) axis.

CONTEXT: Verbascoside (VB), which is found in many medicinal plant families, exhibits biological activities in various diseases. However, its effects on varicocele (VCL)-induced damage remain unknown. OBJECTIVE: To investigate the effects and mechanism of VB on experimental rats with varicocele (VCL)-induced damage. MATERIALS AND METHODS: Sixty sexually mature male Sprague-Dawley (SD) rats were divided into six groups (n = 10): control, control-sham, VCL-vehicle (normal saline), and VCL + VB groups (50, 100, and 200 mg/kg/day, intraperitoneally). After 4 weeks of VB treatment, all animals were sacrificed, and the body and testicular weight, sperm quality parameters, histopathology, antioxidant status, and hormone levels were tested. The levels of gonadotropin-releasing hormone (GnRH) and gonadotropin-inhibitory hormone in the hypothalamus were detected by western blot. RESULTS: Compared with the VCL-vehicle group (41.14%), administration of VB significantly increased the sperm viability (59.29, 65.45, 84.93%). VB groups showed higher Johnson's score (3.57 ± 0.15, 4.71 ± 0.26, 7.93 ± 0.37) than VCL-vehicle group (2.72 ± 0.24). Antioxidant status and hormone levels alterations were also observed. Meanwhile, the mean number of apoptotic tubules (8.15 ± 0.96, 6.61 ± 1.05, 2.17 ± 0.08) and apoptotic index showed a marked decrease. Compared with the VCL-vehicle group (0.21 ± 0.09), the VB groups (0.36 ± 0.07, 0.42 ± 0.06, 0.88 ± 0.10) showed considerable increases in GnRH. DISCUSSION AND CONCLUSIONS: VB has protective effects on reproductive organs and VB may be therapeutically useful in the treatment of varicocele through up-regulation of the HPG axis.

Leaf and Fruit Methanolic Extracts of Azadirachta indica Exhibit Antifertility Activity on Rats' Sperm Quality and Testicular Histology.

BACKGROUND: The world's population is still growing, having an impact on the environment and the economic growth of developing countries; so that, there is a particular interest in the development of new fertility control methods, focused on male contraception. OBJECTIVE: The objective of this study was to evaluate the effect of methanolic extracts of leaf and fruit of Azadirachta indica on sperm quality and testicular histology of Long Evans rats. METHODS: Antifertility effects of a methanolic leaf and fruit extracts of A. indica on 24 male rats were investigated. The animals were randomly divided into two control groups and four treatment groups (n=4). Doses of the leaf and fruit extract were given at concentrations of 100 and 200 µg mL-1. RESULTS: A significant decrease in the viability of sperm cells was observed. The leaf extract at a concentration of 200 µg mL-1 inhibited cell viability compared to the negative control (p< 0.001). The percentage of abnormal cells in leaf extract was shown in 100 and 200 µg mL-1, the conditions at which a higher percentage of morphological irregularities of observed (15% and 16% respectively). The results show that there was cellular detachment in the seminiferous epithelium in the experimental groups treated with methanolic extracts. Sperm death was observed without decreasing the number of sperm. CONCLUSION: The methanolic extracts of Azadirachta indica have a modulating effect on the spermatogenesis of experimental rats through sperm morphological alterations.

Semen quality among young healthy men taking protein supplements.

OBJECTIVE: To study the association between use of protein supplements (PS) and semen quality among young men. DESIGN: Cross-sectional study. SETTING: Not applicable PATIENT(S): We used data from the Fetal Programming of Semen Quality (FEPOS) cohort, which is a subsample of 778 men whose mothers enrolled in the Danish National Birth Cohort 1996-2002. INTERVENTION(S): Semen samples were collected from April 2017 to March 2019. Relative difference in semen characteristics according to self-reported PS use was estimated with negative binomial regression adjusting for lifestyle factors including exercise, body mass index, and use of anabolic steroids, and maternal and paternal factors potentially confounding the association between PS and semen quality. MAIN OUTCOME MEASURE(S): Negative binomial regression yielded the best fit and was used to estimate the percent difference with 95% confidence intervals in semen volume, sperm concentration, total sperm count, proportions of progressive, nonprogressive, and immotile sperm, and percentage of morphologically normal sperm in former and current users of PS relative to never users. RESULT(S): PS was used currently by 28% and formerly by 24% of participants. PS use was not associated with reduced semen quality in terms of semen volume, sperm concentration, total sperm count, morphology, or motility in either crude or adjusted analyses. CONCLUSION: This study showed no association between use of PS and semen quality characteristics. Still, we encourage others to repeat the study, as even a small harmful effect would have a large impact on the population level because of the widespread use of PS among young men.

Combination therapy with antioxidants improves total motile sperm counts: A Preliminary Study.

PURPOSE: Optimal strategies to treat idiopathic male infertility have remained unclear. The aim of this study was to evaluate the effectiveness of combination antioxidant therapy with several vitamins and supplements on semen parameters. METHODS: Thirty-one men with oligozoospermia and/or asthenozoospermia evaluated by a Makler counting chamber were randomly assigned to two treatment groups: a combination of antioxidant supplements (L-carnitine, zinc, astaxanthin, coenzyme Q10, vitamin C, vitamin B12, and vitamin E) and a Chinese herbal medicine, hochu-ekki-to (HE). Serum endocrinological profiles and semen parameters, especially total motile sperm count, were compared between before and after 12 weeks of treatment in both groups. RESULTS: In the supplement group, endocrinological findings were not significantly improved. The semen parameters of semen volume, sperm concentration, and sperm motility were not statistically significantly improved, whereas total motile sperm count was significantly improved. In contrast, none of the endocrinological factors or semen findings were significantly improved by the Chinese herbal medicine although semen concentration, semen motility, and total motile sperm count showed a tendency to increase. CONCLUSION: Because combination antioxidant therapy could improve sperm motility significantly for patients with idiopathic oligoasthenozoospermia, our supplement could be one treatment option for idiopathic male infertility.

Abutilon pannosum stem bark enhances the aphrodisiac activities and spermatogenesis in rat.

Abutilon pannosum (Forst.f.) Schlecht. is used for male sexual performance. In this study, we have investigated aphrodisiac potential of A. pannosum stem bark methanol extract (APM) in rat. Male rats were administered with APM (400 mg/kg) on daily basis for 5, 10 and 15 days. Time interval for mount latency, intromission latency and post-ejaculatory interval was decreased (p < .05) while time of ejaculatory latency, mount frequency, intromission frequency and ejaculatory frequency after 15 days were (p < .05) enhanced as compared to control rats. APM also increased (p < .05) penile erection index, copulatory rate and mount bout against control rats. Total count of spermatozoa was nonsignificantly increased whereas per cent of live spermatozoa and motile spermatozoa were increased (p < .05) in APM treated group after 10 and 15 days. Weight of testes, seminal vesicle, prostate and epididymis, and level of testosterone in serum increased (p < .05) after 10 and 15 days of APM administration to rat. Qualitative characterisation of APM indicated existence of alkaloids, terpenoids, coumarins, cardiac glycosides, phenols, flavonoids, saponins, tannins and sterols. Results of this study indicated aphrodisiac potential of A. pannosum in rat and may be used to enhance sexual performance in human.

[Effect of saw palmetto extract on the reproductive function of male rats with chronic prostatitis].

OBJECTIVE: To investigate the effect of saw palmetto extract (SPE) on the reproductive function of rats with chronic prostatitis (CP). METHODS: Forty male SD rats were equally randomized into groups A (blank control), B (blank control + SPE, C (CP model control) and D (CP model + SPE), and the CP model was made by injection of 1% λ-carrageenan solution into the prostate. The animals in groups A and C were gavaged with normal saline while those in groups B and D with SPE at 0.10 g/kg/d, all for 30 successive days. After drug withdrawal, the rats were mated with female ones in the ratio of 1∶1) and sacrificed 7 days later, their bilateral epididymides collected for detection of sperm count and motility. The numbers of pregnancies and fetuses were recorded and compared among different groups. RESULTS: Compared with the rats in group A, those in group C showed a marked decrease in epididymal sperm motility (ï¼»68.01 ± 1.80ï¼½% vs ï¼»62.59 ± 4.82ï¼½%, P < 0.05), but those in groups B and D exhibited no statistically significant difference (ï¼»67.69 ± 4.06ï¼½% and ï¼»67.93 ± 3.39ï¼½%, P > 0.05). There were no statistically significant differences in the count of epididymal sperm, rate of pregnancy and number of fetuses between group A and the other groups (P > 0.05). CONCLUSIONS: SPE can improve the semen parameters of CP rats, and has no adverse effect on the rate of pregnancy and number of fetuses.

Histomorphology, Sperm Quality and Hormonal Profile in Adult Male Sprague-Dawley Rats following administration of aqueous crude Extract of Solanum nigrum by gastric gavage.

OBJECTIVE: The present study focused on histomorphology, sperm quality, hormonal profile and hematological parameters in adult male Sprague-Dawley rats following the administration of aqueous crude extract of Solanum nigrum by gastric gavage. METHODS: Fourty healthy male adult (12-14 weeks old) Sprague-Dawley rats weighing 200-220g were randomly divided into four groups (A,B,C and D) of ten (n=10) rats each. Group A which served as control were given distilled water 2ml/kg b.wt each, daily for 28 days. Group B, C and D rats were administered 100, 300 and 500mg/kg b.wt each daily respectively for 28 days. The extract was saved with LD50 >5000mg/Kg. Sperm counts, percentage motility, morphology and percentage live sperm, hormonal profile and hematological parameter were quantified; testis, epididymal and general body weights were measured using a weighing scale. The extract was administered once daily for six days within a week via oral gavage. After the last administration, all rats were sacrificed by cervical dislocation, the testis were harvested and fixed in Bouin's fluid for histology processing. RESULTS: Our results revealed an increase in sperm counts, percentage of motility, morphology and percentage of live sperm, blood level of follicle stimulating hormone, Luteinizing hormone and testosterone, hematological parameters, testis, epididymal and general body weights across the groups in a dose-depentant manner. The testis histoarchtecture showed normal cellular composition in their germinal epithelium, with sperm cells in the lumen and a normal interstitium. CONCLUSION: This experiment revealed that aqueous extract of Solanum nigrum bears profertility properties which may be beneficial to those who consume it.

Alteration in apoptotic rate of testicular cells and sperms following administration of Bisphenol A (BPA) in Wistar albino rats.

The aim of the study was to evaluate the effect of Bisphenol A [BPA] widely used as a plasticizer in the formation of polycarbonate plastics and epoxy resins, exposure causing alteration in apoptosis rate, and protective effect of Vitamin E when supplemented with BPA orally. Adult male Wistar albino rats aged 3 months were randomly divided into seven groups: control (olive oil treated) BPA-treated (dose 5, 50,100 µg/100gmBW) and Vitamin E intervention group (dose 5, 50, 100 µg/100gmBW BPA+ Vitamin E dose 4 mg/100gmBW). Animals were sacrificed 3 months later, and blood and tissue samples were collected. Apoptotic changes were analyzed in epididymal spermatozoa and testis tissue by binding of annexin V apoptotic biomarker. A significant decline in the weight of testis, testosterone level, and sperm count was observed. Histopathological and apoptotic changes were observed in testis tissue. In epididymal sperms, the early apoptotic cells were observed by staining of annexin V-conjugated FITC and PI green fluorescence in spermatozoa head which indicated the damage of membrane and late apoptotic cells. These changes reduced significantly in Vitamin E-treated groups though were not found to be comparable to control animals. All these changes were attributed to disrupted spermatogenesis that would interfere with sperm formation. Thus, the study infers that BPA affects the apoptosis process in the testis and epididymal sperm that would interfere with its function and contribute to infertility, whereas Vitamin E-supplemented dose has a protective effect towards these changes, indicating its role in improving male fertility.

Sertoli cell specific knockdown of RAR-related orphan receptor (ROR) alpha at puberty reduces sperm count in rats.

Globally, there is an alarming decline in sperm count. Very often hormonal supplementation fails to restore normal sperm count. Sertoli cells (Sc) present within seminiferous tubules provide appropriate niche and factors required for the differentiation of germ cells (Gc) into mature sperm (spermatogenesis). Functionally compromised Sc may be one of the reasons for failure of hormones to facilitate normal spermatogenesis. Although role of secretory proteins and signaling molecules of Sc has been studied well, role of transcription factors regulating sperm count has not been addressed appropriately. Retinoic acid receptor-related orphan receptor (ROR)-alpha is one of such transcription factors reported in testis but its role in testicular function is not yet known. In a separate study, we found abundant ROR-alpha binding sites on promoter regions of several genes upregulated in pubertal rat Sc as compared to infant Sc. Immunostaining studies also revealed presence of ROR alpha in nucleus of pubertal Sc. We generated a transgenic knockdown rat model expressing shRNA targeted to ROR-alpha under Sc specific promoter, which is transcriptionally active only at and after puberty. ROR-alpha knockdown animals were found to have abnormal association of Sc and Gc, including Gc sloughing and restricted release of sperm. The knockdown animals displayed compromised spermatogenesis leading to significant reduction in sperm count. This is the first report describing the Sc specific role of ROR-alpha in maintaining quantitatively normal sperm output. Identification of various such molecules can generate avenues to limit or reverse an alarmingly declining sperm count witnessed globally in men.

Evaluation of the aphrodisiac potential of a chemically characterized aqueous extract of Tamarindus indica pulp.

ETHNOPHARMACOLOGICAL RELEVANCE: Tamarindus indica is an ingredient in the traditional aphrodisiac formulations in Africa and India. It is also a widely used food ingredient in other tropical countries. AIM OF THE STUDY: The present study was aimed to evaluate the aphrodisiac potential and reproductive safety profile of aqueous extract of Tamarindus indica in male Wistar rats. MATERIALS AND METHODS: The aqueous extract was prepared by maceration of pulp followed by reduction of volume in rotavapor under heat followed by freeze drying. The prepared extract was characterized for contents of total phenol, flavonoid, and saponin. It was also subjected to phytoconstituent analysis using GCMS. Further, the extract was evaluated for acute toxicity study. The aphrodisiac and reproductive toxicity potential were evaluated in animals after grouping them in four with six animals each namely, normal control, standard (Sildenafil citrate, 4mg/kg p.o.) and extract of Tamarindus indica treated groups at two dose levels, 125 and 250mg/kg p.o. The study was conducted for 54 days with daily once dosing of extract and standard. Equal number of females was grouped without treatment for evaluation of parameters of sexual desire (mount frequency and intromission frequency) and parameters of sexual arousal (mount latency and intromission latency). These parameters were evaluated on day 14, 28, 42 and 54. Animals were sacrificed on day 54, testes were removed and studied for histopathological changes. RESULTS: The extract showed 6.6mg gallic acid equivalent/g of total phenol, 2.3mg catechin equivalent/g of flavonoid and 11.6% saponin. Forty chemical constituents were identified by GCMS analysis. In acute toxicity study, the extract was found to be safe till 2000mg/kg p.o. Efficacy study showed significant (p<0.05) improvement in parameters of sexual desire (mount frequency and intromission frequency) and parameters of sexual arousal on all observed days except mount frequency for 125mg/kg on 42nd day and intromission frequency for both doses of tamarind compared to normal control. Improvements in these parameters were comparable to the standard drug. Histopathology study and sperm count suggested an increase in sperm production without any sign of toxicity in testis. Sperm motility significantly (p<0.05) increased in the treatment groups that received extract at 250mg/kg compared to normal control. CONCLUSION: Aqueous extract of Tamarindus indica possessed aphrodisiac activity together with spermatogenic potential.

Walnut leaf extract acts as a fertility agent in male Wistar albino rats - A search for herbal male fertility enhancer.

Background Walnut leaf is one of the many medicinal plants used in folklore as male fertility enhancers. The present work was therefore undertaken with an aim to scientifically validate this claim. As such, we evaluated the effect of the aqueous extract from walnut leaves on biomolecules related to fertility in adult male rats and its mode of action as fertility-enhancing agent. Methods Twenty-five rats were randomly divided into five groups of five animals each; Group 1 served as control and received normal (0.9%) saline only; Groups II, III, IV received 50, 500, 1,000 mg/kg body weight (BW) of T. conophorum leaf extract orally, while Group V served as standard and was given suspension of clomiphene citrate orally at the dose of 1.04 mg/kg/ml BW. The extract and drug were given daily and the experiment lasted for 21 consecutive days. Results The testicular biochemical parameters in treated groups showed significant (p<0.05) increase in lactate dehydrogenase activity activity, Glucose-6-phosphate dehydrogenase (G-6PDH) activity, glycogen content, 3ß and 17ß hydroxysteroid dehydrogenase activities and testicular and epididymal Zn and Se contents with a significant decrease in cholesterol content. A significant increase in testis weight and epididymis weight were also observed. Also, a significant (p<0.05) increase in the level of serum testosterone, luteinizing hormone (LH), follicle-stimulating hormone, sperm count, motility, viability and a decrease in sperm abnormality were observed in the various treated groups when compared with the control group. This increment was concentration dependent, while the extract at the highest concentration showed a more pronounced effect than the standard drug. Also, no sperm DNA fragmentation index was found in all the treatment groups. Photomicrographs from light and scanning electron microscopy showed large fenestrae of interstitial tissue, large fluid space and intact seminiferous epithelium layers fully packed with spermatogenic cells in treated groups than the control group. Conclusions The present study has demonstrated that Tetracarpidium conophorum leaf possesses fertility-enhancing property and have useful effects on spermatogenesis and sperm parameters in rats.

Polar Quassinoids in Standardized Eurycoma longifolia Extract Formulated into a Lipid-Based Solid Dispersion to Improve Rat Sperm Count.

Eurycoma longifolia Jack is popularly sought in Southeast Asian countries for traditional remedies to improve sexual performance and fertility. 13α(21)-Epoxyeurycomanone and eurycomanone, two major quassinoids in a root extract (TAF2) were reported to improve rat spermatogenesis and fertility. Unfortunately, these quassinoids possess low bioavailability because of high aqueous solubility and low lipid membrane permeability. Often, other possible barriers may be P-glycoprotein (P-gp) efflux in the gut and presystemic hepatic metabolism. The present study attempted to solve these problems by formulating a lipid-based solid dispersion (TAF2-SD) of optimized mixture of TAF2 and emulsifiers, which was then orally administered to rats prior to sperm count analysis. The TAF2-SD-treated rats showed significantly twofold (p < 0.001) and fourfold (p < 0.001) higher sperm count than did TAF2-treated and vehicle-treated (control) rats, respectively. The study also demonstrated no significant in vitro ileal absorption changes of the quassinoids by P-gp efflux inhibitors and concentration change or secondary metabolite formation upon in vitro incubation with rat liver homogenates, suggesting that P-gp-mediated efflux and presystemic metabolism were not limiting their bioavailability. Further study on orally TAF2-treated rats confirmed that the area under the curve and bioavailability curve of each quassinoid in the absence and presence of ketoconazole were unchanged. Copyright © 2017 John Wiley & Sons, Ltd.

NMR-based plasma metabolomic discrimination for male fertility assessment of rats treated with Eurycoma longifolia extracts.

Male infertility is one of the leading causes of infertility which affects many couples worldwide. Semen analysis is a routine examination of male fertility status which is usually performed on semen samples obtained through masturbation that may be inconvenient to patients. Eurycoma longifolia (Tongkat Ali, TA), native to Malaysia, has been traditionally used as a remedy to boost male fertility. In our recent studies in rats, upon the administration of high-quassinoid content extracts of TA including TA water (TAW), quassinoid-rich TA (TAQR) extracts, and a low-quassinoid content extract including quassinoid-poor TA (TAQP) extract, sperm count (SC) increased in TAW- and TAQR-treated rats when compared to the TAQP-treated and control groups. Consequently, the rats were divided into normal- (control and TAQP-treated) and high- (TAW- and TAQR-treated) SC groups [Ebrahimi et al. 2016]. Post-treatment rat plasma was collected. An optimized plasma sample preparation method was developed with respect to the internal standards sodium 3- (trimethylsilyl) propionate- 2,2,3,3- d4 (TSP) and deuterated 4-dimethyl-4-silapentane-1-ammonium trifluoroacetate (DSA). Carr-Purcell-Meibum-Gill (CPMG) experiments combined with orthogonal partial least squares discriminant analysis (OPLS-DA) was employed to evaluate plasma metabolomic changes in normal- and high-SC rats. The potential biomarkers associated with SC increase were investigated to assess fertility by capturing the metabolomic profile of plasma. DSA was selected as the optimized internal standard for plasma analysis due to its significantly smaller half-height line width (W h/2) compared to that of TSP. The validated OPLS-DA model clearly discriminated the CPMG profiles in regard to the SC level. Plasma profiles of the high-SC group contained higher levels of alanine, lactate, and histidine, while ethanol concentration was significantly higher in the normal-SC group. This approach might be a new alternative applicable to the fertility assessment in humans through the quantitative metabolomic analysis of plasma without requiring semen. ABBREVIATIONS: TA: Tongkat Ali; LOD: limit of detection; LOQ: limit of quantification; HPLC-UV: high performance liquid chromatography-ultrviolet; PDA: photodiode array; NMR: nuclear magnetic resonance; FID: free induction decay; LC-MS: liquid chromatography-mass spectrometry; GC-MS: gas chromatography-mass spectrometry; HSQC: heteronuclear single quantum coherence; CPMG: Carr-Purcell-Meibum-Gill; VLDL: very low density lipoprotein; HDL: high density lipoprotein; EDTA: ethylenediaminetetraacetic acid; ANOVA: analysis of variance; AMIX: analysis of mixtures; SIMCA: soft independent modeling of class analogy; PCA: principal components analysis; OPLS-DA: orthogonal partial least-squares discriminant analysis; VIP: variable importance plot; AUROC: area under the receiver operating characteristic; TSP: sodium 3-(trimethylsilyl) propionate- 2,2,3,3- d4; DSA: deuterated 4-dimethyl-4-silapentane-1-ammonium trifluoroacetate; ESI: electrospray ionization; TCA: trichloroacetic acid; ACN: acetonitrile; dd H2O: distilled deionized water; FSH: follicle-stimulating hormone; LH: luteinizing hormone; OECD: Organisation for Economic Co-operation and Development.