Juraj Osterman

Gossypol inhibits testicular steroidogenesis

Gossypol, a compound isolated from cottonseed, has been used in the People’s Republic of China as an antifertility agent in men. It was reported that gossypol is highly effective in reducing sperm count. Our study suggests that gossypol inhibits luteinizing hormone, 8-bromo-adenosine 3′,5′-monophosphate-induced testosterone formation, and the conversion of pregnenolone to testosterone in isolated rat interstitial cells. Serum testosterone levels were also significantly reduced in rats after 1 week of treatment with gossypol. Oligospermia and azoospermia in men caused by gossypol may be secondary to decreased testosterone biosynthesis.

The aging Leydig cell. 1. Testosterone and adenosine 3',5'-monophosphate responses to gonadotropin stimulation in rats.

Plasma testosterone levels before and after a single injection of hCG were significantly lower in 24-month old rats than 60--90 day old animals (p less than 0.001). Even with repeated hCG administration for three weeks, plasma testosterone levels of old rats could not be restored to levels present in unstimulated young rats. In response to in vitro LH and 8-bromo-cyclic AMP stimulation, purified young Leydig cells produced significantly higher amounts of testosterone than Leydig cells from old rats. Maximal testosterone formation of the young Leydig cells in response to LH was 42.0 +/- 6.88 ng/10(6) cells, while cells from old rats produced only 16.8 +/- 3.69 ng/10(6) cells (p less than 0.01). However, the dose of LH at which one half maximal response (ED50) occurred was 0.1 mIU/ml for young Leydig cells and 0.05 mIU/ml for old Leydig cells. Basal and 1.0 mIU LH-stimulated cyclic AMP formation were comparable in both groups, but cyclic AMP formation in response to 10 mIU of LH was significantly less in the old rats (p less than 0.05). Present results demonstrate impaired steroidogenic capacity of old rats both in vivo and in vitro. Decreased testosterone response in old rats most likely is the consequence of understimulation of Leydig cells by gonadotropin; however, there appear to be additional intrinsic defects in old Leydig cells.

Direct inhibition of testosterone synthesis in rat testis interstitial cells by ethanol: Possible sites of action

The present in vitro studies using interstitial cells of adult rat testes demonstrated that ethanol inhibits LH- and 8-bromo-cyclic AMP-stimulated testosterone synthesis, pregnenolone- and progesterone-stimulated testosterone synthesis, and basal testosterone synthesis. However, the patterns of inhibition following exposure to 0.22 to 880 or 1100 mM ethanol were different. In general, the inhibition curves for LH-, 8-bromo-cyclic AMP-, pregnenolone- and progesterone-stimulated testosterone synthesis were biphasic, with a gradual slope from 0.22 to 220 mM ethanol, and a sharper slope with concentrations of ethanol greater than 220 mM. Basal testosterone synthesis was reduced only to 74% of control with ethanol concentrations up to 44 mM, and higher concentrations of ethanol reduced testosterone synthesis no further. The effect of ethanol on Lh-stimulated cyclic AMP accumulation showed an even different pattern: some of the lower concentrations of ethanol inhibited cyclic AMP accumulation, while higher levels of ethanol progressively increased cyclic AMP accumulation. These studies demonstrate that isolated interstitial cells are highly sensitive to the direct effects of ethanol; they also suggest that the principle site of ethanol inhibition may be at the level of the smooth endoplasmic reticulum where progesterone is converted to testosterone.

Chronic testosterone cypionate therapy in men with secondary impotence

There have not been studies assessing the effects of chronic testosterone cypionate (TC) therapy on circulating levels of testosterone (T), estradiol (E2), free T, bioavailable T (BAT), luteinizing hormone (LH), and sexual function in impotent men with low T levels. This study was a double-blind crossover using 200 mg of TC or placebo given intramuscularly every 14 days for six injections and the other medication given for six doses. Blood was drawn before each injection. Mean concentrations of T, E2, free T, and BAT were the same on TC or on placebo, but serum LH was significantly suppressed during intramuscular TC. With TC statistically significant improvements in libido and in potency were noted. Five of the men were able to have vaginal sex while taking TC. TC injections every 14 days do not appear to maintain increased T concentrations for 2 full weeks, and other dosage/injection schedules are being evaluated, but there were improvements in libido and potency.

The effects of cytochalasin B on the testosterone synthesis by interstitial cells of rat testis

The present study examined the effects of cytochalasin B on various steps in the luteinizing hormone (LH)-stimulated increase in testosterone synthesis by collagenase-dispersed interstitial cells of adult rat testis. Cytochalasin B at a concentration range of 0.1--50 microM inhibited the LH-stimulated increase in testosterone synthesis in a dose-dependent manner. Both intracellular and medium (released) testosterone levels were reduced, thus indicating that the decrease was not due to the accumulation of testosterone inside the cell as a result of cytochalasin B treatment. Cytochalasin B also inhibited the 8-bromocyclic AMP and pregnenolone-stimulated testosterone synthesis in a similar dose-dependent manner. Cytochalasin B at the two higher doses (10 and 50 microM) also inhibited the LH-stimulated generation of cyclic AMP by interstitial cells. However, this drug had no effect on basal testosterone synthesis except at the highest concentration added. Previous studies on adrenocorticotropic hormone (ACTH)- and LH-stimulated increase in glucocorticoid and testosterone synthesis in adrenal and Leydig cells, respectively, demonstrated that cytochalasin B or anti-actin inhibited the transport of cholesterol into mitochondria. The present studies suggest that cytochalasin B inhibits at least two additional steps in the LH-stimulated increase in testosterone synthesis: (1) the generation of cyclic AMP at the level of the plasma membrane, and (2) the conversion of pregnenolone to testosterone at the level of the smooth endoplasmic reticulum. It remains to be established whether these are direct effects of cytochalasin B, or whether they are mediated by disruption of microfilaments by cytochalasin B.

The aging leydig cell: 2. Two distinct populations of Leydig cells and the possible site of defective steroidogenesis

Using metrizamide gradient centrifugation two populations of Leydig cells were found in both 60-90 day-old and 24 month-old rats. Cells from both Band 2 (B2) and Band 3 (B3) responded to LH stimulation with increased cyclic AMP formation; however, only B3 cells produced significant amounts of testosterone. Cells from both B2 and B3 of the old rats synthesized less cyclic AMP and testosterone than cells from their younger counterparts. In response to LH stimulation, 0.01 - 1.0 mIU/ml, no appreciable difference of cyclic AMP formation could be detected between young and old Leydig cells. Maximal testosterone production occurred when 1 mIU/ml LH was used. Only when LH concentration was increased to 10 and 100 mIU/ml, did young Leydig cells produce significantly more cyclic AMP than old Leydig cells. After addition of 5X10(-7)M of pregnenolone or progesterone to the incubation medium, both young and old Leydig cells produced comparable amounts of testosterone. These results demonstrate no impairment of old rat Leydig cells to synthesize testosterone from pregnenolone and progesterone.

Relationship between inhibition of interstitial cell testosterone synthesis by cytochalasin B and glucose

Abstract We examined the possibility that cytochalasin B reduced LH-stimulated testosterone synthesis of Leydig cells in part by inhibiting glucose uptake. In the presence of 5.6mM glucose cytochalasin B (0.1–50μM) inhibited LH-stimulated testosterone synthesis in a dose-dependent manner, while effects with cytochalasin D or cytochalasin E were not evident until inhibitor concentrations reached 1 or 10μM. Increasing levels of glucose (0–5mM) progressively increased LH-stimulated testosterone synthesis by interstitial cells, while cytochalasin B at 1 or 10μM inhibited testosterone synthesis in a dose-dependent manner at each glucose concentration examined. Higher concentrations of glucose (10– 50mM) partially reversed this inhibition. In the absence of glucose LH-stimulated testosterone synthesis was inhibited only when the cytochalasin B concentration reached 10μM. Cytochalasin B (0.1–5μM) inhibited [ 14 C]-2-deoxyglucose uptake by purified Leydig cells in a dose-dependent manner, while 1 or 10μM cytochalasin D or cytochalasin E had only limited effects. These results suggest that cytochalasin B inhibits Leydig cell LH-stimulated testosterone synthesis by at least two mechanisms: by blocking glucose uptake at lower concentrations (0.1–10μM), while at higher concentrations (10–50μM) it probably blocks microfilament function.

Serum Lipid Physiology and the Influence of Glaucoma Medications

Understanding the effect of serum lipid levels on risk factors for coronary heart disease and how they are influenced by medical therapy may lead to overall better care of the glaucoma patient. Elevated low-density lipoprotein and total cholesterol levels are major risk factors for heart disease. In contrast, high-density lipoprotein (HDL) is protective for heart disease. beta-adrenergic blockers, a class of medicines used to treat glaucoma, may influence serum lipid levels. Oral nonselective beta-adrenergic blockers reduce HDL cholesterol by 19% and increase triglycerides by 20-40%. Furthermore, topical nonselective beta-adrenergic blockers also decrease serum HDL and worsen the total cholesterol/HDL ratio. However, beta-blockers with intrinsic sympathomimetic activity appear to be lipid neutral. At present, there is no clear clinical evidence to indicate that changes in serum lipids with use of topical beta-adrenergic blockers significantly affect the clinical course of the patient. Little information is available for other classes of medicines used topically to treat glaucoma. However, oral preparations of prostaglandins, alpha-adrenergic agonists, angiotensin-converting enzyme inhibitors, and calcium channel blockers do not adversely affect serum lipid levels. Further study is required on newer glaucoma preparations to determine their specific actions on lipid levels. Additionally, further work is required to understand the significance of not only the adverse effect of beta-adrenergic blockers on lipid levels, but their overall effect on long-term cardiac morbidity and mortality.

The aging leydig cell: VII. Cytoplasmic estradiol receptors

Plasma estradiol and cytosolic estradiol receptor levels of testes were determined in a group of young (2-3 months) and old (24 months) Sprague-Dawley rats. Estradiol binding sites for the young rats averaged 5.6 +/- 0.3 fmol/mg protein (x +/- SE, n=12), which was comparable to that of the old rats, 5.7 +/- 0.3 fmol/mg protein (n=12). Using Scatchard analyses, the association constants at equilibrium of estradiol receptor binding of the old and young rats were the same, 6.1 x 10 10 M-1. Plasma estradiol levels were also similar in both groups-19.6 +/- 2.8 pg/ml (n=14) for the young and 19.2 +/- 2.6 pg/ml (n=10) for the old rats. Our results suggest that impaired testosterone biosynthesis in old rats was not due to elevated plasma estradiol levels or to differences in testicular estradiol receptor content.

Case Report: Abnormal Thyroid Function Tests in a Patient and Two Normal Volunteers Treated with Salsalate

There are only three prior reports of abnormal thyroid function tests in patients who have received salsalate, the salicylate ester of salicylic acid. The authors report an elderly clinically euthyroid man who had thyroid function tests suggestive of central hypothyroidism while taking salsalate but whose thyroid tests returned to normal after the drug was discontinued. They also studied thyroid function tests, including free thyroxine (FT4) and reverse (T3), in two normal volunteers who took salsalate 750 mg twice daily for 1 week. In the normal subjects, total T4 and FT4 began to fall within 24 hours after the first dose of salsalate, and remained suppressed for at least 24 hours after the drug was discontinued. This rapidity of effect by salsalate is previously undescribed. There was also a fall in FT4, probably due to the use of diluted serum in the equilibrium dialysis FT4 assay. Because FT4 measurement using diluted serum or equilibrium dialysis may cause falsely low FT4 measurements, the authors believe ultrafiltration may be the only reliable method of measuring FT4 in these patients.