Biochemistry of

Steroid Hormones
Dr. Dilip Kuamr Yadav
Junior resident
Department of Biochemistry
Second year
Content

 Adrenal gland
 Introduction of steroids
 Steroidogenesis –mineralocorticoids,glucocorticoids and
androgens
 Functions and regulation
 Transport and metabolism
 Addison’s disease
 Cohn disease
 Cushing syndrome
 Congenital adrenal hyperplasia
Adrenal gland: suprarenal gland

• There are 2 adrenal glands,pyramidal in shape

• 4 grams each,2-3cm wide and 4-6cm long and 1cm thick
• At the superior pole of both kidneys.
• Each gland is composed of 2 distinct parts:
1. Adrenal Cortex
2. Adrenal Medulla
Adrenal gland
 Introduction
• Steroid hormones are produced by the gonads (ovaries-estrogen and placenta-
progestins) and adrenal cortex(cortisol,androsterone and androgens)

•contains: 17-C-cyclopentanoperhydrophenanthrene

• Made from cholesterol in the smooth endoplasmic reticulum and mitochondria

of endocrine cells.

•Steroid are lipid soluble; thus freely permeable to membranes so not stored in
vesicles.

• Their receptors are located in the cytoplasm target cell.

Steroidogenesis
Adrenal Steroidogenesis
 All steroid hormones fromed from form cholesterol via
pregnenolone through series of reaction

 Enzymes: hydroxylase-requires mol O2(monooxygenase),

NADPH; dehydrogenase ;isomerase;lyase

 Cellular specificity:
• 18 hydroxylase and 18-hydroxysteroid dehydrogenases for MC
synthesis-only in zona glomerulosa
• 17 hydroxylase for hydroxylase for GC synthesis-only in
fasciculata
Adrenal Steroidogenesis
 Cholesterol,mostly from plasma; small portion synthesized in situ
form acetyl-CoA

 Much cholesterol in adrenal esterified and stored in cytoplasmic

lipid droplets

 Upon stimulation of adrenal by ACTH, esterase is activated---

Free cholesterol formed and transported to mitochondria

 ACTH dependent steroidogenic acute regulatory (StAR) protein

essential for transport of cholesterol to inner mitochondrial
membrane--- rate limiting step
Synthesis of steroid
 Cholesterol 1st acted upon by 20,22-desmolase(cytP450 side
chain cleavege enzyme-P450scc)---- forming 21C
pregnenolone: common precursor for all steroid
hormones,ACTH stimulates this step: rate limiting step
Mineralocorticoid Synthesis
 Pregnenolone--- progesterone by two SER enzymes- 3β
hydroxysteroid dehydrogenase(3β-OHSD) and ∆5,4 isomerase.

 21-alpha hydroxylase: Progesterone hydroxylated at C21 to form

11-deoxycortisterone(DOC)-active (Na+ -retaining) MC.

 11- β-hydroxylase: next hydroxylation at C11, produce

corticosterone-has GC activity and weak MC (<5% potency of
aldosterone)

 18-hydroxylase and 18-hydroxydehydrogenase: act on corticosterone

to form 18- hydroxycorticosterone, which is arranged to aldosterone
-CHO group at C18 is
necessary for MC
Mineralocorticoids
 Aldosterone
 Binds to cytoplasmic mineralocorticoids receptor(MR) – DCT,collecting duct,
colon, salivary glands- promote absortion of Na and excretion of K and
Hydrogen ion.

Mineralocorticoid –MR complex relocates to the nucleus

Influences cellular DNA regulating gene transcription

 MR gene encodes 107kDa protein
 Aldosterone stimulates epithelial Na –channel ---via serum and
glucocorticoid-induced kinase and K-ras
 increases expression of mitochondrial ATP-producing genes
stimulate basolateral Na/K pump
Mineralocorticosteroids
Physiology and regulation of
mineralocorticoids
 Average secretory rate is approximately 100-150 μg/day (0.15mg/day)

 Regulation of aldosterone secretion - independent of the regulation of

cortisol and androgens

 Factors that increase aldosterone secretion :

1. Increased Potassium ion concentration (hyperkalemia)
2. Increased activity of the renin-angiotensin system (increased
angiotensin II)
3. ACTH from the anterior pituitary gland (little effect on the rate of
secretion)
• Increased sodium ion concentration very slightly decreases
aldosterone secretion
Renin-Angiotensin Aldosterone
system(RAAS)
Functions of the Mineralocorticoids-
Aldosterone

1. Renal Na+ reabsorption ( action on the principal cells

of the late distal tubule and collecting duct).

2. Renal K+ secretion ( action on the principal cells of

the late distal tubule and collecting duct).

3. Renal H+ secretion ( action on the alpha-intercalated

cells of the late distal tubule and collecting duct).
Glucocorticoids Biosynthesis
 Requires three hydroxylases located in fasciculata and
reticularis: act sequentially on C17,C21 and C11 positions

 17 α hydroxylase (SER): acts upon either progesterone,more

commonly pregnenolone--- 17-hydroxyprogesterone ,17-
hydroxypregnenolone

 21 hydroxylase(SER): 17 hydroxyprogesterone is hydroxylated at C21

to form 11-deoxycortisol

 11β hydroxylase (Mt): hydroxylase 110deoxycortisol at C11 to form

cortisol-most potent natiral GC in humans
 t1/2 of cortisol is 1.5 to 2 hrs
Androgen Synthesis

 Major androgen precursor produced by adrenal cortex:

dehydroepiandrosterone(DHEA)-weak androgen
 Most 17-OH pregnenolone follows GC pathway; small
fraction subjected to oxidative fission and removal of 2-
Cside chain by action of 17,20-lyase(part of same enzyme-
17 α-hydroxylase: dual function protein
 Adrenal androgen production increases markedly if GC
biosynthesis is impeded by lack of any on e of hydroxylases-
adrenogenital syndrome
 DHEA: prohormone ,action of 3β-OHSD and ∆5,4 isomerase it
into more potent androstenedione
 Small amount androstenedione—testosterone(potent adrenal
androgen-mostly in testes)
 Steroid hormone transport
• Are transported by blood from their site of synthesis to their target organs

• Hydrophobic hormone require transport proteins(plasma protein)

• Albumin –nonspecific carrier,carries aldosterone and transthyretin

(prealbumin)

• specific transport molecules (steroid binding globulin-transcortin)-transport

cortisol

• only unbound form can enter the cell

•Steroid and thyroid hormones are 99% attached to special transport proteins
ACTH activity
The biochemical actions of ACTH on the adrenal cortex is summarized as
follows:
(1) Immediate ACTH effects via steroidogenesis activator protein and StAR
(2) Increased cholesterol esterase activity
(3) Decreased cholesterol ester synthesis
(4) Increased cholesterol transport into the mitochondrion
(5) Increased CYP11A binding of cholesterol and
(6) Increased synthesis of pregnenolone.
 Through the action of transcription factors (SF-1), expression of CYP11A,
CYP17, CYP11B1 (but not CYP11B2), and the LDL receptor is increased.
 CYP11B2 is controlled by angiotensin II, separating control of the zona
glomerulosa from the zona fasciculata (CYP11B2 is controlled by ACTH).
 In addition, ACTH and other factors yet not discovered control adrenal
androgen synthesis.
BIOCHEMISTRY OF
ALDOSTREONE,CORTISOL
AND ANDROGENS
Glucocorticoids (Cortisol)
 Glucocorticoids binds to glucocorticoids receptor(GR)- located
at lymphocytes ,hepatocytes and bone
Physiology and regulation of
Cortisol
Continue..
 CRH is released by stress,exercise and hypoglycemia
 It is produced by paraventricular nucleus of the hypothalamus
 Prohormone convertase-1 and PC-2 liberate a C-terminal, 43
aminoacid CRH precursor from 196 amino acid preprohormone
 Peptidylglycine alpha-amidating mono-oxygenase(PAM) removes two
C-terminal residues and adds and amine group ----producing 41 AA
polypeptide CRH
 CRH reaches the anterior pituitary gland through hypothalamic
pituitary portal system
 Corticotrophs express receptor (CRH-R1 and CRH-R2) for CRH----
promotes synthesis, storage and release of corticotropin (ACTH)
 ACTH is also released by ADH,cytokines(IL-1,IL-6 and tumor necrosis
factor)
 Normal cortisol secretion: 6-14mg/m^2 per 24hr
CORTISOL continue…
ANDROGENS
 Dehydroepiandrosterone (DHEA), Dehydroepiandrosterone
sulphate(DHEA-S) and androstenedione -----androgenic effects
through their peripheral conversion to testosterone—which in turns
bind to androgenic receptor(AR).
 At age 7-8years,urinary excretion of 17-ketosteroids increases as an
early sign tha puberty will begin in the coming 3-5 years

 Male – testosterone is more potent androgen

 Female –DHEA and androstenedion more potent
 DHEA-180-1250ng/dL,
DHEA-S—125-169μg/dL
Regulation of Adrenal
Androgens
 Charecterized regulator of androstenedione and DHEA
secretion is ACTH
 Diurnal rhythm in adrenal andrgen concentration
increases in early morning (2 hr before awakening) and
lowest in midnight
 Sympathetic innervtion of zona reticularis may exist and
may regulate adrenal androgen secretion
 Immune regulation has also been proposed
Disorders of Adrenal Cortex

 Addison’s disease
 Hypoaldosteronism
 Cushing syndrome
 Congenital adrenal Hyperplasia
 Functioning Adrenocortical tumors
Addison’s disease
Addison Disease
 Combined mineralocorticoid and glucocorticoid deficiency
 Rare disorder with a prevalence of only 4 to 11 cases per 100,000.8
 Cortisol deficiency is classified as
(1) primary, (2) secondary, or (3) tertiary

 Primary adrenal insufficiency, also known as Addison disease, results

from progressive destruction or dysfunction of the adrenal glands by
(1) an autoimmune process
(2) Some systemic disorder
(3) an inborn error of metabolism (endogenous causes)
(4) by an exogenous cause such as
infection(TB,histoplasmosis,cryptococossis,blastomycosis and
cytomegalo virus,syphillis)
Addison disease
 Secondary adrenal insufficiency, causes:
• tumors • medications • genetics • traumatic brain injury
 SYMPTOMS
• Extreme fatigue
• Weight loss and decreased appetite
• Darkening of your skin (hyperpigmentation)
• Low blood pressure, even fainting
• Salt craving
• Low blood sugar (hypoglycemia)
• Nausea, diarrhea or vomiting (gastrointestinal symptoms)
• Abdominal pain • Muscle or joint pains
• Irritability • Depression or other behavioral symptoms
• Body hair loss or sexual dysfunction in women
DIAGNOSIS
• Complete history collection & Physical examination: perform a
physical exam. Dark patches on your skin might be a clue for your
doctor to consider testing for Addison’s disease. • Assess serum
electrolyte levels : potassium and sodium levels.
• Blood tests: These will be done to measure the levels of sodium,
potassium, cortisol and ACTH in your blood.
• ACTH stimulation test: This tests the adrenal glands’ response after
you are given a shot of artificial ACTH. If the adrenal glands
produce low levels of cortisol after the shot, they may not be
functioning properly.
• X-rays: These may be done to look for calcium deposits on the
adrenal glands.
• Computed tomography (CT scan): Computed tomography uses
computers to combine many X-ray images into crosssectional
views. A CT scan might be done to evaluate the adrenals and/or
pituitary gland. For example, it can show if the immune system has
damaged the adrenal glands or if the glands are infected.
Continue…

Addisonian crisis
• low blood pressure,
• high potassium in the blood, and
• low blood sugar levels.
TREATMENT
• Hydrocortisone pills to replace cortisol.
• If patient also lacking aldosterone, fludrocortisones acetate pills
will be provided.
• If patient are taking fludrocortisones, need to increase salt
intake, especially in hot and humid weather and after exercise.
• In emergencies and during surgery, the medicine is given
intravenously (directly into a vein).
Hyperaldosteronism
Primary Aldosteronism (Conn’s Syndrome)
Causes:
1. Small tumor of the zona glomerulosa cells occurs and secretes large amounts of
aldosterone.
2. In few instances hyperplastic adrenal cortices secrete aldosterone rather than
cortisol.
The effects as mentioned before:
1. Hypokalemia ( occasional muscle paralysis)
2. Slight increase in extracellular fluid volume
3. Slight increase in blood volume
4. Very slight increase in plasma Na+ concentration
5. Almost always hypertention
Diagnostic criteria: Decreased plasma renin concentration (feedback suppression)
Treatment: Surgical removal of the tumor or of most of the adrenal tissue when
hyperplasia is the cause.
Cushing Syndrome
 Endogenous cushing syndrome Condition result of autonomous excessive
production of cortisol
 Exogenous Cushing Syndrome-caused by excessive oral or parenteral
glucocorticoid therapy
Continue…
 Cushing disease: hypersecretion of ACTH by a pituitary microadenoma
is the primary defect that leads to bilateral adrenal hyperplasia and
cortisol overproduction
Pathophysiology
Pathophysiology continue..
Treatment
• Removing an adrenal tumor if this is the cause or decreasing the
secretion of ACTH, if this is possible.

• Hypertrophied pituitary glands or even small tumors, can sometimes be

surgically removed or destroyed by radiation.

• Drugs that block steroidogenesis such as Metyrapone, Ketoconazole and

Aminoglutethimide or that inhibit ACTH secretion ; such as serotonin
antagonists and GABA-transaminase inhibitors, can also be used when
surgery is not feasible.

• Sometimes the only satisfactory treatment is usually bilateral partial (or

even total) adrenalectomy, followed by administration of adrenal
steroids.
Congenital adrenal
hyperplasia
 Most common cause of adreno cortical insufficiency in
newborns
 Types of CAH
1. 21-hydroxylase deficiency (>90%)
 Classical - salt wasting(75%; 1 in 15,000)
- simple virilizing (25%; 1 in 60,000)
 Nonclassic (1 in 1,000)
2. Others
 11Ᏸ-hydroxylase deficiency(3-5 %, 1 in 100,000)
 17α-hydroxylase deficiency / C 17 lyase deficiency (1%)
 3 Ᏸ-hydroxysteroid dehydrogenase deficiency(1%)
CAH
21-alpha hydroxylase
deficiency
Characteristics
Important Lab Investigations

 Glucose/dextrose stick at bedside

 Sr. Electrolytes
 LFT
 Arterial blood gas/serum pH
 Cortisol
 ACTH
 Serum 17-hydroxyprogesterone
 Abdominal Ultrasound – to detect presence of uterus,
cervix and vagina.
 Karyotyping (determine sex chromosome)
DIAGNOSIS
 Diagnosis is based on Elevated 17-OHP levels in
presence of clinical picture.
 Should be performed only in the morning
 In Neonatal period, the levels must be measured after
48 hrs of life due to effect of neonatal surge
 ACTH Stimulation Test : ◦ Measurement of 17 OHP
levels, 1 hr after administration of 250 mcg of IV
Cosyntropin a synthetic ACTH
 Done in Individuals with borderline 17 OHP levels.
Diagnosis

 Elevated Plasma renin activity (PRA) values,

particularly the ratio of PRA to aldosterone, are markers
of impaired mineralocorticoid synthesis.
 Diagnostic Issues
◦ 17 OHP levels are Falsely elevated in stressed and
premature neonates.
◦ Lower levels are observed in neonates exposed to
antenatal steroids
Management
• Glucocorticoids (oral hydrocortisone etc.) 13-18 mg/m²/24hr in 3
divided doses.
Monitoring: serum concentration of adrenal precursors (17-OHP) &
linear growth and skeletal age assessment.
*During stressful state ie febrile ilnesses or surgery, 3x higher dose.
*In severe emergency: intramuscular SC glucocorticoid (Solu-
Cortef)

• Mineralocorticoid therapy (fludrocortisone) at a dose of 0.1-0.2

mg/24hr + sodium chloride supplement 1-2g daily. Monitoring:
serum sodium & potassium, plasma renin activity levels.

• Surgical correction of ambiguous genitalia by 1-2 y/o  normal

development of gender identity.