Vitamin D

Gandham. Rajeev

Department of Biochemistry, Akash

Institute of Medical Sciences &
Research Centre,
Devanahalli, Bangalore,
Kar nataka, India.

eMail: gandhamrajeev33@gmail.com
 Vitamin-D is a fat soluble vitamin

 Vitamin – D is a sterol, it contains steroid nucleus

(Cyclopentanoperhydrophenanthrene ring)
 Vitamin – D function like a hormone

 Forms of vitamin
D:
 Vitamin D in the diet occurs in two forms

 Vitamin D2 (Ergocalciferol)

 Vitamin D3 (Cholecalciferol)
 Ergocalciferol (vitamin D2) is formed from
ergosterol and is present in plants

 Cholecalciferol (vitamin D3) is found in animals

 Both the sterols are similar in structure except

that ergocalciferol has an additional methyl
group(C28) and a double bond

 Ergocalciferol and Cholecalciferol are sources

for vitamin D activity and are referred as
provitamins
CH3
 During the course of cholesterol biosynthesis
7- dehydrocholesterol is formed as an
intermediate
 On exposure to sunlight, 7-dehydrocholesterol
is converted to cholecalciferol in the skin (dermis
and epidermis)
 Dark skin pigment (melanin) adversely
influences the synthesis of cholecalciferol
 The production of vitamin D in the skin is
directly proportional to the exposure to sunlight
and inversely proportional to the pigmentation
of skin

• Excessive exposure to sunlight does not result

in vitamin D toxicity since excess provitamin D3
are destroyed by sunlight itself
 In skin-
sun shine vitamin
7-Dehydrocholestrol
• an intermediate in minor
UV-A
pathway of cholestrol synthesis.
• Found in malpigian layer of epidermis. UV-B
• UV –B light breaks the bond
bet 9—10 in steroid ring
Epidermis
• Form Secosterol
• Cis double bond(bw 5&6C)trans double bon
d dermis
• Form Cholecalciferol
Synthesis vit D

• On skin In plants
7-DehydroCholestrol Ergosterol
UV –B rays Irradiation
Secosterol

Cholicalciferol ErgoCalciferol
(Vitamin D3- provitamin) (vitamin D2-provitamin)
 MITOCHONDRIA, MICROSOMAL
MONOOXYGENASE
Bound to - Vit D binding Protein Need -CYP P-450
-(DBP) -α2- globulin -NADPH
-10 to 65 ng/ml
-biologically inactive
-½ life is 2 to 3 weeks
In mitochondria of PCT
Need - CYP P450
-NADPH
-Ferrodoxin
(Fe-s)
Bound to –
-vit D binding protein(DBP)
-High affinity –
group specific protein
-15 to 60 pg/ml
-biologically active
-½ life 4 to 6 hrs
-excreted thro Bile
 Calcitriol
1,25-diOH-D3
Is a Hormone
 Inactivation
vit D
• By 24-hydroxylase – a
n enzyme expressed i
n most tissues

• Induced by 1,25(OH)2
D – the hormone pro
motes its own inactiv
ation
 Diet from animal sources such as animal
liver contains vitamin D3
 Diet from plant sources contains vitamin
D2
 Absorption: vitamin D2 and D3 are absorbed
from upper small intestine and bile is essential
 Mechanism: vitamin D3 and D2 form mixed
micelles by combining with bile salts (micelles)
 Mixed micelles are presented to mucosal
cells  Absorption occurs by passive transport
 Vitamin D binding globulin: vitamin D is
transported from intestine to the liver by binding
to vitamin D binding globulin

 25 – Hydroxy D3 and 1,25 – dihydroxy D3

are also transported in the blood by binding to
vitamin D binding globulin

Storage:
 25 – hydroxycholecalciferol is the major
storage and circulatory form of vitamin D
 Formation of 1,25 – DHCC is regulated by
the regulation of renal 1 α – hydroxylase
 1 α – hydroxylase activity is increased
by hypocalcemia

 Hypocalcemia stimulates PTH secretion

which, in turn, increases 1 α – hydroxylase
 1 α – hydroxylase activity may be
feedback inhibited by 1,25 – DHCC

 In chronic renal failure, 1 α – hydroxylase activity
is decreased leading to decreased synthesis of
1,25 – DHCC
 The condition leads to renal osteodystrophy
(renal rickets)
 Condition is treated by giving 1,25 – DHCC
preparations
 1 α – hydroxylase deficiency can also occurs as
inherited disorder or due to hypoparathyroidism
 Vitamin D regulates the plasma levels of
calcium and phosphorous
 Plasma calcium levels are regulated by effects
of 1,25 – DHCC on small intestine, kidney and
bone
 It maintains the plasma calcium levels by
increasing absorption of calcium from small
intestine, increasing reabsorption of calcium by
renal distal tubules and increasing mobilization of
calcium from bone
 Calcitriol (1,25 – DHCC) acts at three
different levels to maintain plasma calcium
 Action on
intestine:
 Calcitriol increases the intestinal absorption
of calcium and phosphorus

 In the intestinal cells, calcitriol binds with a

receptor to form a calcitriol-receptor complex
 This complex interacts with a specific DNA
leading to the synthesis of a specific calcium
binding protein

• This protein increases calcium uptake

by intestine
• The mechanism of action of calcitriol is similar
 to that of steroid hormone
 [VDR binds 1,25(OH)2 D
vit D- with 3 times more affinity
FUNCTIONS than other Vit.D metabolites]

Mediates its biologic effects

- Vit D---- bind to----Nuclear Receptor VDR.

- Vit D-VDR complex to Retinoid X Receptor

RXR as a heterodimer

-RXR -Vit D-VDR binds to target DNA sequence

- selectively Stimulates Gene Expression /

Represses Gene Transcription
Hormone –Receptor
complex Vit D- Intestine
Nucleus
Cellular DNA

Increased synthesis
of
specific
calcium-binding
protein

Calbindin
 Action on bone:
• In osteoblasts of bone, calcitriol stimulates calcium
uptake for deposition as calcium phosphate

• Calcitriol is essential for bone formation

• Calcitriol along with parathyroid hormone increase

s the mobilization of calcium and phosphate from t
he bone

• Causes elevation in the plasma calcium and phosp

hate
 Vit D-bone

Vitamin D
Osteoclast Osteoblast
Bone resorption Bone mineralisation

Vitamin D – Co-ordinates the remodelling action

of osteoclasts and osteoblasts.
-10-30% of skeleton remodelled each year
 Action on kidney:

 Calcitriol is also involved in minimizing the

excretion of calcium and phosphate through the
kidney by decreasing their excretion and
enhancing reabsorption
 Effect of vitamin D on kidney

• Stimulates reabsorption of Ca and PO4 in the

distal renal tubules

• PTH conserves only Ca ,selectively excretes PO

4
 Vit D- kidney
Vitamin D
Distal renal tubules
Reabsorption of Ca & PO4

PTH
Distal renal tubules
Secrete Ca ,
Excretes PO4
 kidney
Minimize loss of
calcium
 Regulation of 1,25(OH)2 D
• Induced by:
– Hypocalcemia through PTH
– Hypophosphatemia

• Repressed by :
– Ca2+
– FGF-23 from osteocytes which upregulates 24-hyd
roxylase
 Vit D-
Parathyroid
 Calcitriol is considered as an important
calciotropic hormone, while cholecalciferol is the
prohormone

1. Vitamin D3 (cholecalciferol) is synthesized in the

skin by the UV – rays of sunlight

2. The biologically active form of vitamin D, calcitriol

is produced in the kidney
3. Calcitriol has target organs-intestine, bone and
kidney
4. Calcitriol action is similar to that of steroid hormones
It binds to a receptor in the cytosol and the complex
acts on DNA to stimulate the synthesis of calcium
binding protein

5. Calcitriol synthesis is self-regulated by a feedback

mechanism i.e., calcitriol decreases its own
synthesis

6. Actinomycin D inhibits the action of calcitriol,

calcitriol exerts its effect on DNA leading to the
synthesis of RNA (transcription)
 Vit D-
Diet sources
Egg yolk, Fish oils , Fatty fish , Fortified dairy products

Daily requirment – children – 400 IU / day

< 50 yrs – 200 IU /day
> 50 yrs – 400 to 600 IU /day
 Children - 10 gm/day or 400 IU/day
 Adults - 5 gm/day or 200
IU/day  Pregnency,lactation -10 gm/day or 400
IU/day  Above the age of 60 yrs - 600 IU /day
 Sources of vitamin
D:
 Exposure to sunlight produces
cholecalciferol
 Good sources includes – fatty fish, fish liver
oils, egg yolk etc
 Milk is not a good
source
 Deficiency of vitamin D causes rickets in
children and osteomalacia in adults

Rickets:
 It is a vitamin D deficiency state in
children
 Causes: Dietary deficiency and non-exposure
to sunlight

 Vit D- Deficiency
1) Nutritional deficiency ,
2) Lack of exposure to sunlight,
3) Liver disorder,
4) Renal disorder,
5) Parathyroid deficiency.
 Vit D- Deficiency
Rickets Osteamalacia
Formation of collagen matrix Increased
But incomplete mineralisation demineralization
of pre-existing bone
Soft ,pliable bone
Fractures
Expansion of growth plate osteopenia
 Rickets in children is characterized by
bonedeformities

due to incomplete mineralization
Delay in teeth formation
Causing enlargement and softening of bones
The weight bearing bones are bent to form bow-legs
 Decreased serum calcium
Deformation of muscles: potbelly due to weakness of
abdominal muscles

Biochemical findings:
Decreased serum calcium (9-11mg/dl)
Decreased plasma phosphorous (3-4.5 mg/dl)
Increased plasma alkaline phosphatase (30-130 IU)
 Rickets
Clinical features
• Widened expanded growth plate seen
in long bones and
• Costochondral junctions- leads to swe
lling known as rachitic rosary- X-ray
• Bone deformities- bent bones,
-knock –knee,
-bossing of frontal bones

-pigeon chest, - Harrison’s sulcus

• Proximal myopathy.
Vit D- Def Prior to epiphyseal fusion ,vit D def

Define results in growth retardation asso

Rickets with expansion of growth plate

Blood report
Pl.calcium – low normal
Pl.phosphorus – low normal
Sr.ALP – markedly elevated
• Harrison's sulcus - a  • Pigeon chest:
groove at the lower end • Sternum is raised.
of the rib cage seen in y
oung children / infants
RACHITIC ROSARY
 Vitamin D deficiency in
adults
 Causes: Inadequate exposure to sunlight or
low dietary intake
 Features: Demineralization occurs mainly
in spine, pelvis and lower extremities
 Bowing of the long bones may occur due
to weight of the body

 Flattening of pelvis bones may cause

difficulty during labour
 Vit D- Def Hypocalcemia &
Hypophosphatemia with Vit D def
Osteomalacia -
result in impaired mineralisation of
define bone matrix protein
• In adults Hypocalcemia
Hypophosphatemia
• Impaired mineralization of bone
matrix
INCREASED OSTEOPOROSIS

• Features :
-Bowing of weight bearing
extremities
– Skeletal fractures
 Blood report
Pl.calcium – slightly low
Pl.phosphorus – low
Sr.ALP – markedly elevated

Vit D def ---- never

----severe
Hypocalcemia
No Tetany
 Vit D- Def-Types of Rickets

• Nutritional rickets-
- supplement with Vit.D
• Hypophosphatemic rickets –
-defective renal reab
- Treatment --PO4 + Vit.D
• Vitamin D resistant rickets- fanconi’s syndrome
- defective Reabsorption-
HCo3, Po4, Glu, AA
 Vit D- Def-Types of Rickets

• Renal rickets
– Vit D is present but calcitriol is not
synthesised.
- Trt- calcitriol
• Pseudo Vit D deficiency Rickets –
- mutation in renal 1α hydroxylase.
• Hereditary Vit D resistant Rickets –
- mutation of VDR
 In chronic renal failure, 1 α – hydroxylase
activity is decreased leading to decreased
synthesis of 1,25 – DHCC
 The condition leads to renal
osteodystrophy (renal rickets)
 Condition is treated by giving 1,25 –
DHCC preparations
 1 α – hydroxylase deficiency can also occurs
as inherited disorder or due to
hypoparathyroidism
Biochemical finding
vit D def
1. Sr . 25 (OH) D level
- optimal level > 65 ng/ml
< 15 ng/ml – PTH , Bone density

2. Sr . Calcium - low
3. Sr . Phosphate - low
Therapeutic applications
of Vitamin D
In renal failure
 Used to treat and prevent 20 hyperparathyroidism
- Anti-proliferative effects on parathyroid cells .
- Suppresses transcription of PTH gene .

Research – vit D in cancer treatment.

 Anti-proliferative effects on Keratinocytes,
Breast cancer ,
Prostate cancer cells.
 Vitamin D is stored mainly in liver

 Vitamin D is most toxic in overdoses

 Toxic effects include increased calcium absorption

from intestine, leading increased plasma calcium
(hypercalcemia)

 Hypercalcemia is associated with deposition of calcium in

many soft tissues such as kidney and arteries
 It leads to formation of stones (renal calculi)

 High consumption is associated with loss of appetite,

nausea, increased thirst, loss of weight etc
Hypervitaminosis D
For long periods - Doses >1500IU/day - toxicity
• Vit.D is toxic in high doses
but
• Excessive exposure to sunlight does not result
in Vit.D toxicity
because
Excess D3 is destroyed by sunlight itself
Hypervitaminosis D
For long periods - Doses >1500IU/day - toxicity
Signs & symptoms
-Weakness, Polyuria, Thirst, HT, Weight loss
- Hypercalcemia
-calcification of soft tissues

-metastatic calcification
(calcinosis) in
vascular and renal tissues
THANKYOU
 Harper ’s Biochemistry 25th
Edition.
 Fundamentals of Clinical Chemistry by Tietz.

 Text Book of Medical Biochemistry-A R Aroor.

 Text Book of Biochemistry-DM Vasudevan 

Text Book of Biochemistry-MN Chatterjea 

Text Book of Biochemistry-Dr.U.Satyanarana
 24,25 – DHCC is another metabolite of vitamin
D  It is synthesized in kidney by 24 - hydroxylase
 Calcitriol concentration is adequate, 24 –

hydroxylase acts leading to the synthesis of a

less important compound 24,25 – DHCC

 To maintain calcium homeostasis, synthesis of

24,25 – DHCC is important
 25 – hydroxycholecalciferol 1 –
hydroxylase hydroxylates 25 –
hydroxycholecalciferol at position 1 to
produce 1,25 – Dihydroxycholecalciferol
(1,25-DHCC)
 1,25 – DHCC contains 3 hydroxyl groups (1,
3, 25) and called as calcitriol

 Both hydroxylase enzymes (of liver and kidney)

require cytochrome P450, NADPH and molecular
oxygen for hydroxylation process
 Synthesis of 1,25 – Dihydroxycholecalciferol:
 Active form: the active form of vitamin D is 1,25
– Dihydroxycholecalciferol and is also called as
calcitriol
 Cholecalciferol is first hydroxylated at th

25
position to 25 – hydroxycholecalciferol by a
specific hydroxylase present in liver
 Kidney possesses a specific enzyme, 25
– hydroxycholecalciferol 1 – hydroxylase