Urinary 17-Ketosteroid Conjugates in Hirsutism

Abstract

The principal urinary 17-ketosteroids (17KS) were analyzed in 16 cases of hirsutism; their conjugated forms were separately cleaved, and the liberated 17KS were chromatographed and quantitated. Six were clinically classified as cases of Stein-Leventhal syndrome and the remaining 10 as "idiopathic hirsutism." 1 The following conditions were excluded a priori: adrenal and ovarian tumors, Cushing's disease, and congenital adrenal hyperplasia.Determination of total urinary 17KS in idiopathic hirsutism or in the Stein-Leventhal syndrome has not disclosed conclusive information regarding either the etiology or the pathologic physiology responsible for the appearance of hirsutism in these patients. On the other hand, fractionation of the urinary 17KS has revealed two types of urinary steroid abnormalities in hirsute patients. One abnormality is manifest primarily as an increase in urinary dehydroisoandrosterone 2 both before and after administration of ACTH (2). The second abnormality is an increased urinary level of androsterone and etiocholanolone. In the latter instance some authors have suggested that the adrenal is responsible for the precursors of the increased androsterone and etiocholanolone (3-7), although others have suggested an ovarian origin and still others could not discern the glandular origin of these steroids (8-10). A significant residual level of urinary 17KS after adrenal suppression following administration of 1 A preliminary report of these studies has been presented (1).2 Dehydroisoandrosterone (D): 3,8-hydroxy-5-androsten-17-one. Androsterone (A): 3a-hydroxy-5a-androstane-17-one. Etiocholanolone (E): 3a-hydroxy-5fi-androstan-17-Qne. Isoandrosterone: 3j8-hydroxy-5a-androstan-17-one. 3,8-Etiocholanolone: 3#-hydroxy-5f8-androstan-17-one. 11-Oxy-17-KS (11-0) : sum of 11,6-hydroxyandrosterone, 11 8-hydroxyetiocholanolone, and 11-ketoetiocholanolone. Chorionic gonadotrophin (HCG). Sulfate ester (S). Glucuronoside (G)analogs of cortisol (11-15) favors an ovarian origin of the precursors responsible for the elevation of the androsterone and etiocholanolone values reported.Since the highest blood concentrations of 17KS are represented by those steroids conjugated with sulfuric acid (16), we have restudied the urinary steroids in hirsutism, particularly in order to quantitate the sulfates of 17KS and to discern whether these values more truly reflect the metabolic alteration involved. METHODSTechniques of determination. In order to avoid spontaneous hydrolysis of the conjugated steroids, the 24-hour urine collections were treated with 2 ml of 1 per cent mercurithiosalicylic acid and 5 ml of butanol. The urines were processed immediately or stored at -10°C in the freezer until they could be analyzed.Total 17KS was measured by the method of Jayle, Crepy and Scholler (17). Cleavage of the 17KS conjugates was carried out according to a previously published method (18) which completely separates the glucuronoside from the sulfate fractions by first utilizing the solvolysis technique of Burstein and Lieberman ...

Human Cytosolic 3α-Hydroxysteroid Dehydrogenases of the Aldo-keto Reductase Superfamily Display Significant 3β-Hydroxysteroid Dehydrogenase Activity

“…The function of AKR1C4 is most likely the catabolic inactivation of circulating ⌬ 4 -3-ketosteroids in the liver. There, it works in concert with 5␣/5␤-reductases leading to the formation of 3␣-and 3␤-tetrahydrosteroids that are known to be glucuronidated or sulfated and finally eliminated (21,22). This hypothesis is corroborated by the present study, which demonstrates a lack of stereoselectivity of AKR1C4 in the reductive direction and irreversible epimerization of 3␣-to 3␤-Diol because of negligible oxidation of 3␤-Diol by AKR1C4.…”

“…Reformation of DHT from 3␤-Diol is prevented, because 3␤-Diol is either irreversibly hydroxylated at the C-6 and/or C-7 position or is oxidized to (epi)androsterone (13-20). 3␣-Diol and 3␤-Diol, once formed, are also glucuronidated and sulfated, leading to elimination of the androstanes into the circulation and their final excretion (21,22).In humans, the irreversible NAD ϩ -dependent conversion of ⌬ 5 -3␤-hydroxysteroid into ⌬ 4 -3-ketosteroids is catalyzed by two members of the short chain dehydrogenase/reductase (SDR) family: the bifunctional 3␤-HSD/⌬ 5-4 ketosteroid isomerase (3␤-HSD/KSI) isoforms type 1 and type 2, which express 3␤-HSD and isomerase activity in a single protein (1,2,23). Besides their essential role in the formation of active steroid hormones, the 3␤-HSD/KSI isoforms also catalyze the NAD(H)-dependent in vitro interconversion of 3-keto-and 3␤-hydroxy-5␣-androstanes (24, 25).…”

In VivoStudies on Testosterone Metabolism by Skin of Normal Males and Patients with the Syndrome of Testicular Feminization

“…In all cases, a fraction of urine collected over three days was analyzed according to the previously described procedure (12), including solvolysis, /3-glucuronidase hydrolysis, gradient elution, paper and thinlayer chromatography. In one case (normal male AH), the urine was collected 3, 6, 12, 24, 36, 48 and 60 hours after the administration of labeled precursors and the radioactivity of each urine sample measured after hydrolysis (Fig.…”

The Binding of Androsterone Sulfate, Etiocholanolone Sulfate, and Dehydroisoandrosterone Sulfate by Human Plasma Protein*

“…The 17-ketosteroids appearing in largest quantity in the urine of human subjects are dehydroisoandrosterone, androsterone, and etiocholanolone.1 These steroids are not excreted to any appreciable extent in unconjugated form, but are conjugated as sulfates and glucuronides (1,2). Dehydroisoandrosterone is excreted mainly as its sulfate congener, whereas androsterone and etiocholanolone appear in the urine mainly as glucuronides.…”

“…Dehydroisoandrosterone is excreted mainly as its sulfate congener, whereas androsterone and etiocholanolone appear in the urine mainly as glucuronides. Nevertheless, the latter two compounds are also excreted as sulfates, and the concentration of these sulfates in urine may equal or exceed that of dehydroisoandrosterone sulfate (1,2). It has been established that dehydroisoandrosterone sulfate (D-S) is the main 17-ketosteroid in human peripheral plasma (3)(4)(5).…”