Evaluation of the antiosteoporotic potential of Tinospora cordifolia in female rats.

UNLABELLED: The available courses of therapy to osteoporosis in menopausal women are limited by several side effects generated. A need therefore arises to explore herbal alternatives that are effective and safe. OBJECTIVE: Present animal studies were conducted to investigate the potential of Tinospora cordifolia (TC) ethanolic stem extract as an antiosteoporotic agent. METHODS: Three-month-old female Sprague-Dawley rats were either ovariectomized (ovx) or sham operated and treated with vehicle (benzyl benzoate:castor oil; 1:4), E(2) (1 microg/day) or TC (10, 50, 100 mg/kg b.wt) subcutaneously for 4 weeks. At the end of experiment bone mineral density of tibiae was measured by quantitative computer tomography. Serum was analyzed for the activity of alkaline phosphatase and levels of osteocalcin, cross-laps and lipids. Uterus and mammary gland were processed for histological studies. RESULTS: Ovx rats treated with TC (10 mg/kg b.wt) showed an osteoprotective effect as the bone loss in tibiae was slower than ovx controls. Serum osteocalcin and cross-laps levels were significantly reduced. All the above effects of TC were much milder than those produced by E(2). Alkaline phosphatase activity was higher in TC treatment groups. Total cholesterol and LDL levels remained unaltered but HDL levels were significantly lowered with TC (50 mg/kg b.wt) treatment. Uterus and mammary gland showed no signs of proliferation after treatment with TC extract. CONCLUSION: TC extract showed estrogen like effects in bone but not in reproductive organs like uterus and mammary gland. Thus, this study demonstrates that extract of T. cordifolia has the potential for being used as antiosteoporotic agent.

Spearmint induced hypothalamic oxidative stress and testicular anti-androgenicity in male rats - altered levels of gene expression, enzymes and hormones.

Mentha spicata Labiatae, commonly known as spearmint, can be used for various kinds of illnesses in herbal medicines and food industries. One of the prominent functions of this plant extract is its anti-androgenic activity. The present study investigated the probable correlation between oxidative stress in hypothalamic region and anti-androgenic action of this plant's aqueous extract on rats. Decreased activities of enzymes like superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase in hypothalamus of treated rats indicated spearmint induced oxidative stress. Further RT-PCR and immunoblot analysis demonstrated the decreased expression of some of the steroidogenic enzymes, cytochrome P450scc, cytochrome P450C17, 3beta-Hydroxysteroid dehydrogenase (3beta-HSD), 17beta-Hydroxysteroid dehydrogenase (17beta-HSD) and other related proteins like, steroidogenic acute regulatory protein, androgen receptor and scavenger receptor class B-1. Further, in vitro enzyme assays demonstrated depressed activities of testicular 3beta-HSD and 17beta-HSD enzymes. Histopathology indicated a decreased sperm density in cauda epididymis and degeneration of ductus deference. Our study suggested that spearmint probably induced oxidative stress in hypothalamus resulting in decreased synthesis of LH and FSH which in turn down-regulated the production of testicular testosterone through the disruption of a number of intermediate cascades.

Phospholipase-C sensitive GPI-anchored proteins of goat sperm: possible role in sperm protection.

The role of glycosylphosphatidylinositol (GPI)-anchored sperm proteins in reproduction has been investigated. SDS-polyacrylamide gels (PAGE) analysis of goat sperm (Capra indica) indicated that several GPI-anchored proteins were released by phosphatidylinositol-specific phospholipase-C (PI-PLC) treatment. The distribution of this category of PI-PLC-sensitive GPI-anchored proteins on the surface of sperm was examined by indirect immunofluorescence. The fluorescence microscopic study clearly demonstrated that the PI-PLC-sensitive GPI-anchored proteins are confined predominantly to the head region of goat sperm. Further experiments were conducted on intact and PI-PLC treated sperm in order to decipher the function of GPI proteins. Co-incubation of sperm with peritoneal macrophages led to the enhanced phagocytosis of PI-PLC treated sperm by macrophages compared with the untreated intact sperm. Transmission electron micrographs of the macrophages acquired from the phagocytosis assay are provided to corroborate the same. From the results obtained it is inferred that one or more of the PI-PLC-sensitive GPI-anchored proteins on the sperm surface could act as protection factor(s) that shield the sperm from macrophages.

A comprehensive evaluation of the reproductive toxicity of Quassia amara in male rats.

Chloroform extracts of the bark of Quassia amara in different dilutions was used to assess its impact on the male reproductive system of albino rats. Single daily intramuscular injections of the extract for 15 days resulted in a significant reduction in the weight of testis and epididymis but not that of the seminal vesicles and prostate (all lobes). A marked decrease in the sperm count, motility, viability was also observed in sperm collected from the cauda epididymis of treated animals. A number of abnormalities like double heads, double tails, detached heads and fragile tails were frequently seen. Epididymal alpha-glucosidase activity was drastically reduced. However, prostatic acid phosphatase activity and citric acid levels and seminal vesicle fructose concentrations remained unchanged following treatment. Thus, it appears that the prime site of action is at the level of both the testis and the epididymis. Examination of the blood showed that cell counts and hemoglobin levels were in the normal range. Bilirubin, SGPT, SGOT, protein and urea were also not altered by the herbal extract. From the selective action on the male reproductive tract we suggest that the chloroform extract of the bark of Quassia amara has potential for use as an antifertility agent.

N-acetyl beta-D-glucosaminidase is not attached to human sperm membranes through the glycosylphosphatidyl inositol (GPI)-anchor.

AIM: The mode of anchorage of N-acetyl beta-D-glucosaminidase (NAGA) on human ejaculated sperm was investigated. METHODS: Sperm plasma membrane was prepared by discontinuous sucrose gradient centrifugation from human sperm. NAGA was solubilized from these membranes by two detergents: octyl-glycoside and triton X-100. In separate studies, the release of the enzyme from the sperm membrane preparation by phosphatidylinositol specific phospholipase C (PI-PLC) was also examined. RESULTS: NAGA activity was detected on sperm membranes isolated from human ejaculates. The pattern of the enzyme solubilization by detergents indicated that the enzyme was an integral protein of sperm membrane. NAGA was not released from the sperm membranes by PI-PLC treatment. CONCLUSION: The evidence presented strongly suggests that human sperm membrane bound NAGA is not attached via the GPI anchor.

Androgenic action of Tinospora cordifolia ethanolic extract in prostate cancer cell line LNCaP.

AIM OF THE STUDY: Recently, Tinospora cordifolia (TC) was shown to affect prostate growth in rats. It is not known whether this is a direct effect of TC or whether it is induced by altered hormone release. To investigate the actions of TC on the prostate, human LNCaP cells were exposed to an ethanolic extract of TC. MATERIALS AND METHODS: LNCaP cells were incubated with the test substances for 48 h. Proliferation was measured by MTT test and prostate-specific antigen (PSA) secretion was determined with ELISA. RESULTS: TC showed a dose-dependent stimulation of proliferation of LNCaP cells. Co-incubation with the anti-androgen flutamide (FLU) reversed the TC-induced stimulation of PSA secretion. CONCLUSIONS: The reference compound dihydrotestosterone (DHT) caused a significant increase of growth of LNCaP cells. Similarly, TC stimulated proliferation of these prostate cells. The anti-androgen FLU reversed the increase of PSA release caused by either DHT or TC. Thus, we suggest that TC may contain androgenic compounds, which appear to act via androgen receptor (AR).

Characterization of biofilm formed on intrauterine devices.

PURPOSE: Intrauterine device (IUD) is one of the most convenient contraceptive procedures used by women of Asian and African countries. Previous surveys have revealed that 75% of the IUDs recovered from patients suffering from reproductive tract infections (RTIs) were covered with a consortium of microbes. This study was designed to characterize these microbes and recommend remedial measures. METHODS: Quantitative measurement of biofilm formation was assessed by a microtitre plate assay on 86 samples of microorganisms dislodged from IUDs of patients with RTIs. Susceptibility of biofilm to various antimicrobial agents was also quantified. Scanning electron microscopy (SEM) was used to scrutinize the microorganisms adherent to IUDs. RESULTS: The organisms associated with IUDs were predominantly composed of Staphylococcus aureus (16%), Staphylococcus epidermidis (18%), Pseudomonas aeruginosa (5%), Escherichia coli (27%), Neisseria gonorrhoeae (2%), Candida albicans (20%) and Candida dubliniesis (12%). SEM studies indicated that these organisms were organized into biofilms. Studies on the in vitro adherence pattern by crystal violet staining on 96 well microtitre plates revealed that the biofilms were stably established after 60 hours. These biofilms are resistant to an array of antibiotics tested. CONCLUSION: Biofilm formation may be one of the major causes for persistent infection and antibiotic resistance in IUD users.