Suramin: an inhibitor of the final steps of the mineralocorticoid pathway?

The authors used incubated adrenal mitochondria to study the in vitro effect of suramin, an antiparasitic drug, on the transformation of corticosterone and 18-hydroxycorticosterone into aldosterone. The results show that, under conditions preserving membrane integrity, the "impermeance" of suramin meant that concentrations similar to the plasma-levels reached in treated patients induced only slight inhibition of the final intramitochondrial steps in aldosterone synthesis. However, suramin strongly inhibited mitochondrial respiration. The inhibition of two intramitochondrial mechanisms (respiration and steroid synthesis) suggests that the effect of suramin involves partial inhibition of metabolic intermediate carriers. The inhibition of the activity of various extramitochondrial enzymes involved in intermediate metabolism, suggests that the inhibition of steroid biosynthesis can be explained only on the basis of an extramitochondrial action of suramin. The action of suramin must, therefore, primarily and directly affect extramitochondrial steroid synthesis and only indirectly affect intramitochondrial steroid synthesis as a result of an impact on the reducing equivalent supply. However, even if suramin does not bind to cytochrome P450 11 beta which catalyzes the final steps of aldosterone biosynthesis pathway, this does not imply that suramin has no direct effect on steroid synthesis within the mitochondria, in addition to its toxic effects, particularly if the cell structure is disrupted (as is often the case in tumor tissues).

Hypoaldosteronism accompanied by normal or elevated mineralocorticosteroid pathway steroid: a marker of adrenal carcinoma.

In order to find a biochemical marker to assist the physician in the difficult differential diagnosis between malignant and nonmalignant adrenal tumors, plasma levels of the mineralocorticosteroids (deoxycorticosterone, 18-hydroxydeoxycorticosterone, corticosterone, 18-hydroxycorticosterone, and aldosterone) were determined. The same method (RIA which is preceded by a crucial separation step) was used to measure all these steroids including aldosterone. The subjects included 15 adults presenting various clinical signs of adrenocortical tumors (histopathologically: 6 with adrenal carcinoma, 1 with a history of adrenal carcinoma, 1 with adrenal metastasis from other forms of cancer, 6 with adenoma, and 1 with hyperplasia). The results show that both presurgery and during a recurrence of adrenal carcinoma, hypoaldosteronism occurs which contrasts with the normal or even elevated levels of some aldosterone precursors. In the 7 cases of adrenal cortical carcinoma, this dysfunction of the aldosterone pathway was detected regardless of the impairment of the other steroidogenesis pathways, whereas it was never found with a nonmalignant tumor. Despite the limited number of cases so far available, these findings suggest that detection of abnormalities of the aldosterone pathway, and particularly the detection of hypoaldosteronism by an assay method involving a crucial steroid separating step, could contribute to a differential diagnosis between benign and malignant adrenocortical tumor and between adrenal metastasis and other forms of cancer.

Cortisol and testosterone: inhibitory agents of the mineralocorticoid pathway. Is there a physiopathological meaning in adrenal tumors?

The authors incubated adrenal mitochondria to study the in vitro action of cortisol and testosterone on the transformation of corticosterone and 18-hydroxycorticosterone into aldosterone. The results show that cortisol at concentrations of 5 x 10(-6) and 10(-4) M inhibit the conversion of corticosterone into aldosterone by 23.6 to 90%; testosterone 5 x 10(-5) and 10(-4) M inhibit the reaction by 78.4 and 87.2%, respectively. The inhibition of the conversion of 18-hydroxycorticosterone into aldosterone is 12.5 to 91% by cortisol with concentrations ranging from 5 x 10(-7) to 5 x 10(-5) M and testosterone 5 x 10(-5) and 10(-4) M inhibits the reaction by 87.3 and 91%, respectively. Aldosterone (10(-8) and 10(-6) M) does not inhibit aldosterone biosynthesis from corticosterone or 18-hydroxycorticosterone. It thus appears that cortisol and testosterone have an effect on the aldosterone biosynthesis pathways in mitochondria. This action may be located at the binding site of the cytochrome P450 11 beta, which catalyzes all hydroxylation steps in the mineralocorticoid biosynthesis pathway. Because cortisol and testosterone may interfere with aldosterone biosynthesis, and since functional zonation is expected in adrenal carcinomas, the presence of these steroids in substantial amounts could explain the very low plasma aldosterone level usually observed, in adrenal carcinomas studied in our laboratory.

[Organ transplantation. One of the biochemical reasons for the race against the clock]. / Transplantations d'organes. Une des raisons biochimiques de la course contre la montre.

The authors have studied the subcellular functioning of human adrenal glands removed from subjects in a stage IV coma. The present study has a two-fold interest: on the one hand, it offers biochemical information on a key element in the intermediate metabolism (namely, the mitochondrial energetic metabolism, occurring in a fragile tissue which, under a state of shock, is primarily affected; the results obtained on such type of tissue may therefore be inferred to other organs); on the other hand, it allows a wider approach of the adrenal biochemical mechanisms during a stage IV coma. The mitochondrial fraction was obtained by differential centrifugation carried out immediately after organ removal. The steroid synthesis, studied using radioactive precursors, turned out to be similar to that found in other mammals. Respiratory characteristics, determined by polarography with a Clark oxygen electrode, at 37 degrees C, were satisfactory: respiratory intensity was 77.25 +/- 12.16 nanomoles O2/min/mg mitochondrial protein in the presence of succinate 15 mM and respiratory control was 1.93 +/- 0.15 in the presence of ADP 37 microM. The respiratory chain functioned in a classical manner: rotenone 25 microM did not inhibit respiration in the presence of succinate 15 mM, while it did with L-malate 15 mM. In the presence of succinate 15 mM, the respiratory intensity was inhibited at 87.4 percent and 76.7 percent by KCN 0.01 microM and antimycin A 0.09 microM respectively; with DNP 85 microM, it was multiplied by 5. However, the value of the P/O ratio was low (0.24 +/- 0.04). Under the present conditions, this may highlight the difficulty to synthetize ATP whenever neither the functioning nor the regulation of the multi-enzymatic complex accounting for oxygen consumption are affected. This result clearly confirms that the shortest possible delay between organ removal and transplantation is crucial, as the renewal of cell structures requires energy. These fundamental research studies account for the major concerns of reanimation teams. This also raises the issue of the role of fundamental researchers within a transplant surgery team.

[18-Hydroxycorticosterone, 18-hydroxydesoxycorticosterone. Why, when, how to determine plasma levels in some adrenal pathologies]. / 18-Hydroxycorticostérone, 18-hydroxydésoxycorticostérone. Pourquoi, quand, comment en déterminer les taux plasmatiques dans certaines pathologies surrénaliennes.

The authors review some current ideas concerning the role of 18-hydroxylated corticosteroids as mineralocorticoids themselves and as possible precursors of the principal mineralocorticoid, aldosterone. In particular, the physiological and pharmacological agents affecting their secretion are discussed together with a description of the methods used for their analysis in plasma in the department of Clinical Biochemistry Pitié-Salpétrière. Finally, the value of these assays in the differential diagnosis of mineralocorticoid hypertension and inborn errors of corticosteroid biosynthesis is assessed and the constraints on sampling technique listed.

Verapamil directly inhibits aldosterone synthesis by adrenal mitochondria in vitro.

The action of verapamil, a calcium channel blocker, on the last step of aldosterone biosynthesis (transformation of 18-hydroxycorticosterone into aldosterone) was studied using duck adrenal mitochondria in the absence of regulatory factors. Results show that 10(-5) M verapamil inhibits the transformation of 18-hydroxycorticosterone into aldosterone by 52.8%. Moreover, our findings show that verapamil induces only a slight inhibition of respiratory capacity without action on respiratory control and does not displace 18-hydroxycorticosterone from cytochrome P450 11 beta which catalyses the reaction. Thus, this study does not explain the mechanism of inhibition induced by verapamil on the last step of aldosterone synthesis but it is of interest to note, for clinical use, that this inhibition is not linked to regulatory factors of aldosterone production. Since primary hyperaldosteronisms are characterized by their independence vis-á-vis regulatory factors, administration of verapamil may be particularly interesting for treatment of primary hyperaldosteronisms.