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An electron microscopic study of the changes induced by oestrogens on the lymph-node post-capillary venules.

The changes induced in the lymph-node post-capillary venules of male and female mice, 1, 2, 4, 7, 10 and 14 days after oestrogen administration were studied in 1 micron sections and with the electron microscope. The endothelial cells of the post-capillary venules are higher in female than in male control mice. On the first and second days after Hexoestrol administration there is an increase in the number of ribosomes and polyribosomes lying free in the cytoplasm of the endothelial cells. These cells were significantly increased in height between day 2 and day 7, in both male and female Hexoestrol-receiving mice, remaining at high levels thereafter. The increase in height was accompanied by an increase in the Golgi elements which occupied the major part of the cytoplasm of the endothelial cells. Lysosomes, multivesicular bodies and signs of phagocytosis were more often seen. In contrast, the numbers of lymphocytes passing through the post-capillary venule wall were significantly decreased, probably due to a direct effect of Hexoestrol on the thymus. The relation of an oestrogenic effect on the ultrastructure of the post-capillary venules to the sex differences seen in the immune response is discussed. The possibility that this may constitute a part of a general oestrogenic effect on the reticuloendothelial system is also considered.

Animals↗

Ultrastructural effects of sex hormones and infection on lymph node post-capillary venules.

The ultrastructure of the lymph node post-capillary venules (PCV) was studied in control, gonadectomised and gonadectomised-Hexoestrol treated male and female mice, infected or not with Toxoplasma gondii. The percentage of the cytoplasmic area of the endothelial cells occupied by Golgi apparatus was estimated by quantitative stereology. The endothelial cells of the PCV are higher and contain more Golgi elements in female than male controls. Oophorectomy decreases the percentage of Golgi apparatus and the height of the endothelial cells while orchidectomy does not affect the former but increases the latter. The endothelial cells react to Hexoestrol similarly in male and female mice, by increasing their height and becoming more heavily structured, with an effect particularly on the Golgi apparatus. All groups of mice infected with Toxoplasma gondii, show increased numbers of lymphocytes passing through the wall of the PCV. The endothelial cells of the latter show increased numbers of ribosomes both free and attached to rough endoplasmic reticulum. Golgi elements are decreased. In view of these findings the mechanisms for selective lymphocyte migration through the walls of the PCV are discussed.

Animals↗

Hepatotoxicity of the synthetic oestrogen hexoestrol in the female rat.

The synthetic oestrogen hexoestol is hepatotoxic to the female rat. Twenty-five animals were treated daily with hexoestrol at a dose of 60 mg/kg. Twenty-five more acted as controls. Five animals from each group were killed after 4, 12, 20, 30 and 40 days of treatment to permit serial evaluations of liver histopathology to complement serum biochemical investigations. There were no mortalities during the study and only modest external signs of toxicity. All the treated rats showed reductions in body weight and appetite and gains in liver weight. This latter effect was due to both cellular hypertrophy and hyperplasia. The principle finding of the liver histopathology was fatty change. This affected scattered individual cells, mainly in the midzonal region. All of the serum parameters of toxicity--which consisted of bilirubin, total protein, albumin, glycoprotein, alpha 2-macrofoetoprotein, and alkaline phosphatase--indicated liver impairment. Most of these also showed time-related changes consistent with an initial phase up to Day 20 of marked liver dysfunction superseded by a more adaptive phase thereafter. The hepatotoxicity described here seems sufficient to explain blood clotting defects observed elsewhere in oestrogen-treated rats.

Alkaline Phosphatase↗

Anti-proliferative effects of 1,2-diphenylethane oestrogens and anti-oestrogens on human breast cancer cells.

The anti-tumour activities of 1,2-diphenylethane oestrogens (hexoestrol and orthohexoestrol) and anti-oestrogens (metahexoestrol, tetramethylHES, and metatetramethylHES) were studied on the human MCF-7 and MDA-MB-231 breast cancer cell lines. On the E2R-positive MCF-7 cell line, all test compounds exhibited a dose-dependent inhibition of cell proliferation, but no correlation between anti-proliferative activity and binding affinity for the E2R was found. Tested on the E2R-negative MDA-MB-231 cell line, metahexoestrol also showed dose-dependent inhibitory effects, but higher concentrations were necessary than on the MCF-7 cell line. From this it is concluded that the anti-proliferative effect is specific and at least partially mediated via the E2R. Combination of metahexoestrol (10(-6)M) with E2 (10(-9) to 10(-7)M) gave no rescue effect. It is therefore suggested that this compound might be useful for therapy in the presence of high oestrogen levels, i.e. in pre-menopausal patients. The test compounds (10(-8) to 10(-6)M) could rescue the inhibitory effect of tamoxifen (10(-6)M) in a dose-dependent manner, except in the cases of metahexoestrol (10(-6)M) and tetramethylHES (10(-6)M). The latter compound exhibited a strongly additive effect at this concentration.

Breast Neoplasms↗

3,4-bis(3'-hydroxyphenyl)hexane--a new mammary tumor-inhibiting compound. Studies on the mechanism of action on the DMBA-induced, hormone-dependent mammary tumor of the rat.

Biochemical properties, such as the activities of eight carbohydrate-metabolism-linked enzymes and four acid hydrolases, and histological characteristics of growing and regressing DMBA-induced mammary tumors of the SD-rat after ovariectomy or treatment of the host with hexesterol, tamoxifen, and 3,4-bis(3'-hydroxyphenyl)hexane were determined. Significant differences were found between growing and regressing tumors regardless of the treatment animals had been subjected to. Only few differences in biochemical parameters could be found within the group of tumors regressing due to the applied therapy. The histological signs of regressing tumors were very diverse, but no phenomenon typical of a specific treatment could be found. It cannot be decided whether the partial antiestrogen, 3,4-bis(3'-hydroxyphenyl)hexane unfolds its antimammary tumor activity by means of its estrogenic or its antiestrogenic potency. The hypothesis that estrogens inhibit mammary tumor growth by directing neoplastic cell metabolism toward secretion is not supported by these findings.

9,10-Dimethyl-1,2-benzanthracene↗

Effects of hexoestrol on the contractility of the isolated, blood-perfused canine papillary muscle and of the guinea-pig ileum.

The effects of hexoestrol were studied on the blood flow and the tension development of the isolated, blood-perfused papillary muscle of the dog, as well as on the contractions of the guinea-pig ileum induced by acetylcholine, histamine or BaCl2. Hexoestrol caused an increase in coronary blood flow and a negative inotropic effect in a dose-dependent manner. High doses of hexoestrol produced a loss of contractility of the papillary muscle without significant alterations of the amplitude and shape of the electrical activity recorded extracellularly. Hexoestrol had a strong non-selective relaxant effect on the guinea-pig ileum and was 10-20 times more potent than papaverine in antagonizing contractions induced by the 3 agonists. These results indicate that hexoestrol probably acts by blocking certain steps in the process by which extracellular and/or superficially bound Ca ions move to the contractile machinery in cardiac and in smooth muscle cells. Thus, it may bear some resemblance to the so-called calcium antagonists.

Acetylcholine↗

Differential effects of chloroquine and of several other amphiphilic cationic drugs upon rat choroid plexus.

Several cationic amphiphilic drugs, each of which is known to induce generalized lipidosis in rats, were compared with respect to their cytological effects on rat choroid plexus epithelium. Chloroquine induced large cytoplasmic vacuoles, whereas the other drugs (quinacrine, 4,4'-diethylaminoethoxyhexestrol, chlorphentermine, iprindole, 1-chloro-amitriptyline, clomipramine) caused formation of lamellated or crystalloid inclusions as usually seen in drug-induced lipidosis. The ultrastructure of the chloroquine-induced vacuoles suggested storage of water-soluble materials (polar lipids and/or non-lipid materials) in addition to non-water-soluble polar lipids.

Amitriptyline↗

Inhibition of acid esterase in rat liver by 4,4'-diethylamino-ethoxyhexestrol.

The effect of 4,4'-diethylamino-ethoxyhexestrol (DH) on acid esterase in rat liver was studied in vivo and in vitro. The acid esterase activity in the livers of rats treated with 0.125% DH for 1 week was found to decrease more than 60% as compared with that in untreated rats. The addition of DH to the incubation medium caused considerable inhibition of the acid esterase activity in lysosome from untreated rat liver, and the inhibition type appears to be noncompetitive. The acid lipase activity in rat liver lysosome was also inhibited by DH. Some antihistamic agents and chloroquine also inhibited the acid esterase activity in rat liver lysosome.

Acid Phosphatase↗

Peripheral nervous system affection in experimental lipidosis induced by 4,4'-diethylaminoethoxyhexesterol.

A picture of generalized phosphoglyceride and cholesterol storage was induced, in keeping with literary data, by the experimental administration of 4,4'-diethylaminoethoxyhexesterol to rats. An asset of this model lies in the discovery that considerable storage occurs in the peripheral nervous system in contrast to the CNS, whose resistance to hexesterol is generally known. The significance of this finding is briefly discussed.

Animals↗