Ascorbic Acid and Alpha-Tocopherol Contribute to the Therapy of Polycystic Ovarian Syndrome in Mouse Models.

Polycystic ovary syndrome (PCOS) affects up to 10% of women within reproductive ages and has been a cause of infertility and poor quality of life. Alteration in the oxidant-antioxidant profile occurs in PCOS. This study, therefore, investigates the contribution of ascorbic acid (AA) and alpha-tocopherol(ATE) on different PCOS parameters. The mifepristone and letrozole models were used, and young mature female mice were randomly assigned to groups of six per group. On PCOS induction with either mifepristone or letrozole, mice were administered AA and ATE at doses ranging from 10-1000mg/kg to 0.1-1000 mg/kg in the respective models. Vaginal cytology, body weights, and temperature, as well as blood glucose, testosterone, and insulin levels, were measured. Total antioxidant capacity and malondialdehyde levels were analyzed. Determination of gene expression of some reactive oxygen species and histomorphological analysis on the ovaries and uteri were performed. At the end of the experiments, AA and ATE restored reproductive cycling, with AA being more effective. AA and ATE increased fasting blood glucose but had no significant effect on serum insulin levels. AA decreased testosterone levels, but ATE caused slight increases. AA and ATE both increased total antioxidant capacity and decreased malondialdehyde levels. AA and ATE also slightly upregulated the mRNA expressions of catalase, superoxide dismutase, and heme oxygenase 1 mainly. AA and ATE also decreased ovarian weight and mostly resolved cysts in the ovaries and congestion in the uterus. This study has shown that AA and ATE are beneficial in the therapy of PCOS.

Thyme (Thymus vulgaris [Lamiaceae]) Leaves Inhibit Contraction of the Nonpregnant Mouse Uterus.

Dysmenorrhea is painful menstrual periods, which affects 25% of women within reproductive age and has a prevalence of 67.2-90.0%. Current treatment has several adverse effects and can be ineffective once the pain is initiated. Thymus vulgaris traditionally used for pain management was investigated in this study for its activity on uterine contraction in the nonpregnant uterus, as a parameter for dysmenorrhea. The dried leaves of T. vulgaris were macerated in water, and the resulting aqueous extract was investigated on the isolated mouse uterus. Parameters investigated included spontaneous contractions, oxytocin-induced contractions, and high potassium chloride (KCl; 80 mM)-induced tonic contractions. Mass spectrometric analysis of the thyme extract was also performed using liquid chromatography-high-resolution Fourier Transform mass spectrometry. Thyme extract inhibited the amplitude and frequency of spontaneous and oxytocin-induced uterine contractions. It also inhibited KCl-induced tonic contractions. The activities observed suggest that T. vulgaris inhibits uterine contractions through blockade of extracellular voltage-gated calcium channels. Secondary metabolites detected included compounds belonging to chlorogenic phytochemical class and flavonoids, which are known to have activities on extracellular calcium blockade. This study has shown that aqueous T. vulgaris extract, also known as thyme, inhibits contractions of the nonpregnant uterus and can be a lead plant in the drug discovery process for the management of dysmenorrhea.

Justicia flava Leaves Exert Mild Estrogenic Activity in Mouse Models of Uterotrophic and Reproductive Cycle Investigations.

This study aimed to assess and determine the estrogenic activity of the leaf extract of Justicia flava (JF) in mice, which may interfere with its therapeutic use in female reproduction. The uterotrophic assay (UTA) utilizing 20 days old female mice and the reproductive cycle assay (RCA) utilizing adult female mice were used in this study. All administrations were performed orally. Reproductive organ and blood samples were collected the day after last administration of JF for histology and hormone analysis. Other parameters such as organ weight, temperature, body weight, and reproductive cycles were analyzed. Our study showed that for UTA, JF increased uterine weights slightly, which were nonsignificant but more pronounced at the highest dose of 1000 mg/kg. JF did not induce vaginal opening, which is a sign of puberty onset. JF also had minimal effect on organ morphology and caused a slight increase in serum estrogen. For RCA, JF did not significantly alter body weight and temperature although an upward trend in temperature was observed. JF did not disrupt cycling significantly (P > .005) compared with estrogen (the positive control drug used). JF also did not significantly alter uterus morphology except at 1000 mg/kg where some increase in the number of glands and cell activity were observed. JF has mild estrogenic activity and will not interfere with reproductive functions at lower doses (10-100 mg/kg) during therapy, but high doses (up to 1000 mg/kg and above) may cause some alterations. Our data, therefore, suggest that JF is a useful candidate in the management of female reproductive health issues at lower doses.

Disruptions in the female reproductive system on consumption of calcium carbide ripened fruit in mouse models.

This study investigated for the first time the outcome of ingestion of calcium carbide-ripened fruit on some female reproductive parameters. A set of unripe mature bananas ripened with calcium carbide (CCRB) and another set ripened via non-artificial means (NARB) were fed orally to prepubertal female mice for three days using the uterotrophic assay procedure. A distilled water group and oestradiol group (10 mg/kg) were also assigned. Food intake, body weights, vaginal openings and cytology were analysed. Samples of blood, uteri, ovaries and cervices were additionally collected and analysed. Increased serum oestrogen level and uterus weight were detected in the CCRB and oestradiol treated groups. Histopathology showed increased numbers of myometrial cells, presence of secondary follicles and regressing corpus lutea as well as thickened cervix epithelia which were evidence of oestrogenic disruptions. This study has shown that consumption of fruits ripened with calcium carbide negatively alters the female reproductive physiology, accelerates puberty onset and increases serum oestrogen levels. Caution must therefore be exercised by fruit sellers in the use of calcium carbide and policies set in place for strict regulation of its use worldwide.

Tocolytic activity assessment of the methanol leaf extract of Justicia flava Vahl (Acanthaceae) on mouse myometrial contractility and preliminary mass spectrometric determination of secondary metabolites.

ETHNOPHARMACOLOGICAL RELEVANCE: The leaves of Justicia flava are traditionally used in the South of Nigeria to prevent preterm births. AIM OF THE STUDY: In this study, the activity of the methanol leaf extract of J. flava (JF) was investigated on uterine contractility in non-pregnant and pregnant isolated mouse tissues. MATERIAL AND METHODS: The effects on spontaneous, oxytocin, and KCl-induced contractions were determined. The effects in calcium-free media were also determined. Possible mechanisms of activity were investigated using receptor and channel modulators. Mass spectrometric analysis was additionally performed on the leaf extract to identify secondary metabolites. RESULTS: JF was observed to inhibit spontaneous, oxytocin and high KCl-induced uterine contractility. JF also inhibited contractions in Ca2+-free media. JF was found to exert its inhibitory effect via interaction with inositol triphosphate and ryanodine receptors and also through modulation of K+- channels. Lignans and alkaloids were identified with the lignans being the most abundant in JF. CONCLUSION: JF has been shown to potently inhibit uterine contractions in non-pregnant and pregnant isolated mouse uterus. The inhibitory activity of JF has been shown to occur via blockade of extracellular and intracellular calcium entry and these effects may be due to the lignans identified in - JF. JF has therefore been shown in this study to be a lead plant in the discovery of new drugs with uterine inhibitory activity.

Amelioration of Escherichia coli-induced endometritis with ascorbic acid in non-pregnant mouse models.

PROBLEM: Infection-induced endometritis is associated with infertility. The outcome with oral antibiotics remains poor. This study therefore investigates the role of ascorbic acid in resolving endometritis. METHOD OF STUDY: Animals with established oestrus cycles were inoculated with Escherichia coli. Two days post-inoculation, the animals were administered ascorbic acid (10, 100 and 1000 mg/kg) and amoxicillin (500 mg/kg) for 5 days. Other groups included water only and E. coli inoculated with no treatment. Body temperatures, weights and vaginal cytology were examined. On the sixth day, after anaesthesia, blood samples were obtained for haematological analysis. Uterine organs were weighed, ex-vivo functionality analysed and histopathological analysis performed. RESULTS: Ascorbic acid (AA) (100 and 1000 mg/kg) regularized the cycle of the endometritic animals comparable to amoxicillin. AA (1000 mg/kg) and amoxicillin, significantly decreased (P < .05) the endometritis-induced increase in uterine weights, restored the endometrial architecture and significantly (P < .05) normalized uterine contractions to control values. Improved haematological profiles were additionally observed on treatment with ascorbic acid (100 and 1000 mg/kg). CONCLUSION: AA compared favourably with amoxicillin in endometritis management, suggesting that AA can be used in the management of infection-induced endometritis, normal cycling and normal uterine function.

A role of alpha-tocopherol and phylloquinone in the modulation of uterine contractility and reproductive function in mouse models.

BACKGROUND AND AIM: Alpha-tocopherol has been implicated in reproduction processes, and deficiency of phylloquinone has been associated with serious complications in pregnancy. This study was therefore aimed at investigating the effects of phylloquinone and alpha-tocopherol on uterine contractility and female reproductive function using mouse models. MATERIALS AND METHODS: Both in vivo and ex vivo animal models were employed and designed to assess changes on uterine contractility and reproductive functions in the non-pregnant uterus. The effect of alpha-tocopherol and phylloquinone on spontaneous uterine contractions, oxytocin-induced uterine contractions (11.82nM) and high KCl-induced tonic uterine contractions (80mM) were assessed. The effect of subcutaneous administration of alpha-tocopherol (10mg/kg) on reproductive hormone levels and reproductive tissues were also determined. RESULTS: Alpha-tocopherol increased the force of contractions while phylloquinone decreased the force of uterine contractions. Plasma levels of luteinizing hormone (P<0.01), estrogen (P<0.01) and progesterone (P<0.001) were elevated in the presence of alpha-tocopherol after 6 days subcutaneous administration. CONCLUSIONS: Alpha-tocopherol and phylloquinone have been shown to directly modulate uterine contractility and reproductive function and may contribute to the management and treatment of reproductive disorders.