Effect of Polyphenols on Production of Steroid Hormones from Human Adrenocortical NCI-H295R Cells

Abstract

Modulating steroid hormone levels is a curative and preventive measure for Cushing's syndrome, aldosteronism, and various stress-triggered symptoms. Polyphenols have been reported to inhibit steroidogenic enzymes such as 3β-hydroxysteroid dehydrogenase (3β-HSD) and aromatase. However, evidence for their inhibitory effects is fragmentary because it has been determined in studies with small groups of steroid hormones. To investigate the effects of steroids on complete steroidogenic pathways, comprehensive analysis of steroid hormones is necessary. Here we cultured forskolin-stimulated NCI-H295R, a human adrenocortical carcinoma cell line, in the presence of a polyphenol and employed GC-MS to simultaneously determine the levels of nine steroid hormones (pregnenolone, progesterone, deoxycorticosterone, aldosterone, 17α-hydroxyprogesterone, dehydroepiandrosterone, androstenedione, testosterone, and estradiol) in cell culture supernatant. We found that daidzein, genistein, apigenin, hesperetin, naringenin, and eriodictyol significantly reduced deoxycorticosterone and androstenedione levels (p<0.05), suggesting inhibition of 3β-HSD by these polyphenols. Apigenin was more potent than other polyphenols in increasing the levels of pregnenolone and 17α-hydroxyprogesterone, suggesting that it inhibits cytochrome P450 (CYP) 17 and CYP21, as well as 3β-HSD. Real-time reverse transcription polymerase chain reaction showed that apigenin significantly downregulated the expression levels of 3β-HSD, CYP17, and CYP21 mRNA ( p<0.05). This is the first study to demonstrate the inhibitory effects of apigenin on CYP17 and CYP21.Key words apigenin; cytochrome P450 (CYP)17; CYP21; pregnenolone; 17α-hydroxyprogesterone; polyphenol Steroid hormones are secreted by the adrenal cortex and play pivotal roles in stress response, immune response, antiinflammatory actions, carbohydrate metabolism, protein catabolism, and electrolyte homeostasis.1) Disrupting the secretion of these hormones has a serious impact on human health. For example, in Cushing's syndrome, hyperproduction of cortisol results in redistribution of fat to the face (moon face), hypertension, central obesity, and striae distensae. In cases of aldosteronism, hyperproduction of aldosterone induces renal reabsorption of sodium and water, which leads to a rise in circulating blood volume and hypertension. In addition, under stressful conditions, such as those related to deteriorating socioeconomic situations, chronic work-related stress, anxiety, and depression, the neuroendocrine response is upregulated, and the resulting cortisol hyperproduction triggers insulin resistance. Furthermore, the hypothalamic-pituitary-adrenal axis is hyperactivated under chronic stress, which in turn induces abdominal obesity, thereby leading to metabolic syndrome. 2)Agents and supplements that modulate steroid hormone levels play pivotal roles in treating the above diseases and conditions. Several food constituents have already been reported as steroid hormone inhibitors. For example, water-soluble l...

Apigenin inhibits growth of the Plasmodium berghei and disrupts some metabolic pathways in mice

“…In our study, intraperitoneal administration of apigenin in mice affected the classical and alternative pathways of bile acid metabolism by increasing chenodeoxycholic acid, lithocholic acid, taurohyodeoxycholicacid, hyodeoxycholic acid, and cholesterol derivatives. Apigenin was shown to significantly down‐regulate the expression of 3β‐HSD (3β‐hydroxysteroid dehydrogenase), CYP17A1 (17α‐hydroxylase), and CYP21A2 (21‐hydroxylase) at mRNA levels (Hasegawa et al, ). 3β‐HSD is an enzyme that catalyzes the biosynthesis of progesterone from pregnenolone, 17α‐hydroxyprogesterone from 17α‐hydroxypregnenolone, and androstenedione from dehydroepiandrosterone (DHEA) in the adrenal gland (Lachance et al, ).…”

“…Flavonoids have also been shown to modulate key enzymes in adrenal steroidogenesis, affecting steroid hormone biosynthesis in the mineralocorticoid, glucocorticoid, and adrenal androgen pathways (Schloms & Swart, 2014). The estrogenic properties of apigenin have already been described (Hasegawa, Nakagawa, Sato, Tachikawa, & Yamato, 2013). Apigenin is able to trigger steroidogenic acute regulatory (StAR) gene expression at the transcriptional level and steroid hormone production in mouse Leydig cells by blocking the thromboxane A2 receptor and reducing the DAX-1 (dosage sensitive sex reversal-adrenal hypoplasia congenita critical region on the X chromosome, gene-1) protein, a transcriptional repressor of StAR gene expression (Li et al, 2011).…”

Antisecretive and Antitumor Activity of Abiraterone Acetate in Human Adrenocortical Cancer: A Preclinical Study

“…Liquid Cromatography-Tandem Mass Spectrometry. Steroid extraction was performed as described in [19] and is reported in the Supplementary Materials and Methods. Analysis was carried out using a highperformance liquid chromatography (HPLC) system (1260 Series Agilent Technologies; Santa Clara, CA, USA) equipped with a reversed-phase Hypersil Gold C8 (100 x 3mm 3um) analytical column (Thermo AA was administered i.p (196 mg/Kg) daily for 16 days (qdx16); control mice received the vehicle (2.5%benzyl alcohl, 97.5% safflower oil).…”

Effect of Polyphenols on 3-Hydroxy-3-methylglutaryl-Coenzyme A Lyase Activity in Human Hepatoma HepG2 Cell Extracts

“…It has been reported that certain polyphenols inhibit digestive enzymes such as lipase, α-amylase, α-glucosidase, 6) and steroidogenic enzymes, 7) as well as liver microsomal glucuronidation of estrone and estradiol.…”

“…5,6) We recently reported that apigenin, a flavonoid found in several herbs including parsley, thyme, and peppermint, inhibits steroidogenic enzymes of cytochrome P450 (CYP)17 and CYP21, as well as 3β-hydroxysteroid dehydrogenase. 7) HMG-CoA lyase in hepatic cells is a rate-limiting enzyme catalyzing the cleavage of HMG-CoA to acetoacetate, and thus inhibition of this enzyme results in reduced acetoacetate production, in other words, impaired ketogenesis. Based on the notion that inhibition of HMG-CoA lyase possibly prevents ketoacidosis and the identification of therapeutic targets, we examined the inhibitory effects of polyphenols on HMG-CoA lyase activity in cellular extracts of human hepatoma HepG2 cells.…”