Effect of catecholestrogen administration during adriamycin-induced cardiomyopathy in ovariectomized rat.

The therapeutical beneficial effect of estrogen-derived metabolites or catecholestrogens is controversial. These molecules are produced during estrogen therapy based on 17-beta-estradiol treatment. The metabolization of 17-beta-estradiol is carried out in brain, kidney or liver, and triggers different products such as 2- and 4- hydroxyestradiol (2OH and 4OH). These products have shown antioxidant properties against oxidative stress (OS) in several experimental models. Different noxious side effects related to those metabolites have also been observed upon estrogen therapy. In this sense, catecholestrogens seem to be implicated in tumoral and mutagenic process after long treatment with estrogens substitutive therapy. In our study, we have verified that 2OH and 4OH have antioxidant and cardioprotective effects against adriamycin (AD)-induced cardiomyopathy in ovariectomized (OVX) rats. Catecholestrogens diminished the lipid peroxides and carbonyl protein (CO) content, and different enzymes related to cell injury (creatinine kinase, lactate dehydrogenase, aspartate aminotransferase, alanine aminotransferase) in cardiac tissue from OVX-, AD-, and OVX+AD-treated rats. All these changes were correlated to a recovery on reduced glutathione (GSH), glutathione peroxidase (GPx), superoxide dismutase (SOD) and catalase (CAT) in heart tissue. The present study showed that 2OH and 4OH reduced all the parameters related to OS, antioxidant depletion and cardiac injury in OVX rats treated or not with AD.

Differential effects of estrogen metabolites on bone and reproductive tissues of ovariectomized rats.

The effects of 17 beta-estradiol and the important estrogen metabolites, 2-hydroxyestrone (2-OHE1) and 16 alpha-hydroxyestrone (16 alpha-OHE1) on bone, mammary gland, and uterine histology, and on blood cholesterol were investigated in ovariectomized growing rats. Rats were treated with 200 micrograms/kg of body weight/day of each of the test compounds for 3 weeks. Ovariectomy resulted in uterine and mammary gland atrophy, increased body weight, bone turnover and tibia growth, and hypercholesterolemia. 17 beta-estradiol treatment prevented these changes, with the exception that this high dose of estrogen did not prevent hypercholesterolemia. 2-OHE1 had no effect on any of the measurements. 16 alpha-OHE1 resulted in bone measurements that did not differ from the 17 beta-estradiol-treated rats and prevented the increase in serum cholesterol. In contrast, 16 alpha-OHE1 resulted in increases in uterine weight, uterine epithelial cell height, and mammary gland cell proliferation that were significantly less than the 17 beta-estradiol treatment. These findings demonstrate that 16 alpha-hydroxylation of estrone results in tissue-selective estrogen agonistic activity, whereas 2-hydroxylation resulted in no measured activity. Furthermore, they suggest that factors that modulate the synthesis of these metabolites could selectively influence estrogen target tissues.

Effect of intraventricular administration of catechol estrogens on catecholamine content in various brain regions.

Male rats treated for 7 days with either 2-hydroxyestradiol, 4-hydroxyestradiol or 2-hydroxyestrone had significantly lower striatal dihydroxy phenylacetic acid (DOPAC) levels when compared to the control group. After 14 days of treatment, groups which were treated with estradiol, 2-hydroxyestradiol or 4-hydroxyestradiol had significantly higher striatal dopamine levels and had gained significantly less weight when compared to the control group. These results indicate that estradiol and the catechol estrogens may act to reduce the activity of striatal dopaminergic neurons.

Influence of catecholestradiol on short-lived corpora lutea in beef cows.

The influence of intrauterine administration of catecholestradiol (4-hydroxylated estradiol) on lifespan of the initial postpartum corpus luteum was evaluated in suckled beef cows. In experiment 1, postpartum cows (n = 23) were untreated (CONTROL) or received intrauterine infusions (0700 and 1700 hr) of either vehicle (SAL) or catecholestradiol (CATE; 4 micrograms) from day 15 to 22 (day 0 = parturition). Blood samples were collected three times weekly (day 15 to 100) and analyzed for progesterone. In experiment 2, cows received twice daily intrauterine infusions of either vehicle (n = 18), or catecholestradiol (n = 19), from day 25 +/- .5 to day 30 +/- .5. Following the final infusion, calves were temporarily weaned from all cows for 48 hr. At the end of the 48 hr weaning period, cows in each infusion group received either an i.m. injection of 1,000 IU hCG (SAL+hCG, n = 9; CATE+hCG, n = 9) or no further treatment (SAL, n = 9; CATE, n = 10). Blood samples were collected daily for 21 d following calf removal and 3 times weekly through 100 d postpartum. In both experiments, the initial postpartum elevation in peripheral progesterone concentrations was characterized as either a short (< 5 d) or extended (> 8 d) luteal phase. In experiment 1, postpartum anestrous interval (60 +/- 3.4 d) and incidence of short luteal phases (77%) were similar among CONTROL, SAL and CATE treatments. In experiment 2, luteal phases were induced within 10 d of onset of weaning in 90, 100, 56 and 60% of cows in SAL+hCG, CATE+hCG, SAL and CATE treatments, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Influence of subchronic administration of catechol estrogens on the formation of reactive oxygen species in rat liver microsomes.

Metabolic pathways of estrogens are the formation of catechol estrogens (CE; 2- and 4-hydroxy-estrogens), redox cycling of CE and free radical generation, mediated through cytochrome P450 (P450) oxidase/reductase activity. In previous investigations subchronic administration of estrogens showed prooxidative and antioxidative activities in rat liver microsomes (BARTH et al. 1999). To find out whether or not catechol metabolites are responsible for prooxidative activity, we checked 2- and 4-hydroxy-estradiol (2OH-E2 and 4OH-E2) and the non-catechol metabolite 6alpha-hydroxy-estradiol (6alpha-OH-E2) for formation of reactive oxygen species in liver microsomes of 30-day-old male Wistar rats after 5 days treatment (1, 10 mg/kg b. wt. orally, once a day). The results were compared with those after treatment of the rats with estradiol (E2), estradiol valerate (E2V) and ethinylestradiol (EE2). In liver homogenates glutathione and lipid peroxides were determined, in microsomes NADPH-Fe++-stimulated lipid peroxidation (LPO), H2O2 generation and lucigenin (LUC) and luminol (LUM) amplified chemiluminescence (CL) were investigated. In liver 9000 x g supernatants monooxygenase activities were measured. The two catechol estrogens did not show any antioxidative activity, whereas 6alpha-OH-E2 significantly diminished lipid peroxides in the liver as well as LPO and LUM-CL in liver microsomes. Among estrogens, only EE2 showed antioxidative activity. Both CE inhibited ethoxycoumarin O-deethylation. Peroxidative activity as enhanced LUC-CL was found after 2OH-E2 (1 mg/kg b.wt.) and E2, but 10 times higher doses of both CE did not change LUC-CL. Microsomal H2O2 generation was enhanced by E2, E2V and both CE, not by 6alpha-OH-E2. The lower level of H2O2 enhancement caused by CE in comparison to E2 and E2V together with unchanged LUC-CL after high CE doses did not unequivocally prove the CE to be mainly responsible for the prooxidative activities of E2 and E2V in liver microsomes, at least in 30-day-old male rats. Unchanged GSH in the liver after CE administration supports this hypothesis.

[Mechanism of the suppressive effect of catechol estrogen on the preovulatory LH surge].

The administration of 100 micrograms 2-hydroxyestrone (2-OHE1) or 2-hydroxyestradiol-17 beta (2-OHE2) at 0900h or 1000h in the morning of proestrus into normal 4-day cycling rats results in suppression of the preovulatory LH surge in the afternoon of that day. The LH-RH content of the median eminence in control rats decreases sharply in the afternoon from elevated noon levels. However, the catechol estrogen-treated rats do not show this decrease. These results indicate that the catecholestrogens block the preovulatory LH surge by preventing the release of LH-RH from the median eminence, but do not interfere with the synthesis and accumulation of LH-RH. Furthermore, injections of non-uterotropic 2-OHE1 twice at 30 minutes intervals into ovariectomized rats fail to affect the LH tonic secretion by 180 minutes after the initial injection. These data indicate that catechol estrogen interferes with the brief neuronal triggering phase necessary for LH-RH release, but does not affect the LH tonic secretion which is an estrogen-independent process.

Effect of continuous intraventricular estrogen or catechol estrogen treatment on catecholamine turnover in various brain regions.

The effect of 7-day i.v.t. administration of catechol estrogens (CE) or estrogens (5 micrograms/day) on the catecholamine turnover rate of various brain areas was examined in ovariectomized rats. Norepinephrine turnover was increased significantly in the hypothalamus and cerebral cortex by estradiol treatment but not by any CEs tested when compared to control values. However, the turnover rate of dopamine in the cerebral cortex was increased compared to control values only by the 2-hydroxyestrogens (2-hydroxyestradiol and 2-hydroxyestrone) and estradiol was without effect. Only estrogens and CEs with physiologically significant estrogen receptor binding affinities (17 beta-estradiol, moxestrol, 2-hydroxyestradiol and 4-hydroxyestradiol) decreased the turnover rate of dopamine in the corpus striatum compared to control values. Estrogens (17 alpha-estradiol and 2-hydroxyestrone) which are weak ligands for the estrogen receptor did not affect striatal dopamine turnover. In addition, body weight gain measured during estrogen treatment was reduced by CEs and estrogens which have significant estrogen receptor affinities. These results suggest that the CEs may play a role in central modulation of catecholaminergic function by estrogens either through direct actions of the catechol moiety or activation of estrogen receptors.