MICRONUTRIENTS HNSC 7211 Prof.

Kathleen Axen Spring 2014

The sunshine vitamin

Is it really a vitamin?
An organic compound Natural component of food Is essential in the diet
>60% of requirement is produced in skin, conditions permitting

For normal physiologic function

0.025 g (25 ng) of D2 or D3 = 1 IU DRI ~400IU or 10 g

Dietary Vitamin D
D2 Corn oil 9 IU/100g D3 Fish liver oils 10,000 IU/100g Salmon 220 IU/100g Butter 35 IU/100g Poultry 80 IU/ 100g Poultry skin 900 IU/ 100g
Fortified dairy products (D2 or D3 can be used)
Humans do not distinguish between forms (birds do)

400 IU/ quart

Absorption
Bile required, formation of micelles Passive diffusion ~ 50% bioavailability Most (>90%) in chylomicrons Non-esterified Transferred to Vitamin D Binding Protein in plasma Vit D remaining in chylomicron is taken up by liver but no storage in liver (~1/4 of dose in bone)

Synthesis in skin
UV B 285-315 nm (UVA > 315nm, does not cause sunburn, UVC < 285nm) Photoisomerization (no enzyme) 7-dehydrocholesterol secreted by sebaceous glands on surface absorbed through skin to Malpighian-deep-layer beneath stratum corneum D3 and metabolites synthesized Excess D3 converted to byproducts
All UV types can be mutagenic and cause collagen destruction in skin

How much sun exposure do you need for adequate vitamin D synthesis?
Holicks Rule: MED over of body yields 1,000 IU Or 6-10% of body surface exposed until mildly sunburned (MED) DRI=200 for most categories (1 IU/.025 g) How many g?

Amount synthesized depends on

Thickness of skin, greater in
Adults vs. children Men vs. women People of African vs. Asian or European ancestry

Amount of pigment (melanin) in skin Use of sunscreen, clothing Time of day (highest 10 am 2pm) Season, latitude (in winter no D synthesis in Denver, NYC, Toledo, Beijing)

Vitamin D Binding Protein

Synthesized in liver
DBP transports D2 or D3 and metabolites in plasma Highest affinity for 25 hydroxy vit D 50% of D bound to DBP Mobilizes vit D formed in skin (so that form is more efficiently used)

Other binding proteins

Cytoplasmic
Vitamin D receptor in nucleus (VDR)
Binds to 1, 25 dihydroxy D Necessary for the genomic effects of vit D

25 hydroxy cholecalciferol (25 OH D)is the major form in blood; reflects dietary intake

25 hydroxylase

Regulation of vit D Metabolism

25 hydroxylase (liver) driven by vit D conc 1 hydroxylase (kidney)
Zinc dependent by PTH (due to low Ca), prolactin, low phosphate by FGF 23 (fibroblast growth factor)from osteocytes
FGF23 is by calcitriol It phosphate reabsorption in kidney and absorption in SI

by inflammation Regulated differently in other tissues

Forms calcitriol =1,25 dihydroxy vit D

 Maintaining normal blood levels of Ca2+ is crucial for survival (1 M)

It is the job of PTH and vitamin D-related mechanisms Bone is the storage place for Calcium; plasma Ca2+ concentration is a far greater priority than bone mineral content or density Bone mineral is resorbed to provide Ca for the blood

Calcium
Ca 2+
Ion in solution Part of inorganic salts
Buffers Bone mineral Tooth mineral (enamel)

Bioavailability
Increased by Solubility
Acid form: e.g. calcium gluconate

Lactose Ascorbic acid Interaction with phosphorylated proteins

casein lactalbumin

Bioavailability
Decreased by Oxalate Dietary fiber Phytate

phytate

Ethanol Fat malabsorption (soap formation) Tannins, polyphenols (flavonoids)

Calcium absorption:
Paracellular: across tight junctions between intestinal epithelial cells
Big effect of concentration gradient, time in intestine (length, important in ileum, maybe colon)

transcellular: transport into, across and out of intestinal epithelial cells

Several models proposed, with evidence > 1 model may operate, or combinations

Paracellular Transport

Tight junctions in SI epithelia

Calcium Transcellular Transport Mechanisms

Ca enters via TRPV6

Crosses via
Cytosol? ER? Vesicles?

Exits via Ca ATPase (PMCA)

Entry: apical surface

Ca2+ channel protein TRPV6 (transient receptor potential of the vanilloid type) Expression is increased by calcitriol (VDR-mediated,slow) Rapid effect of calcitriol: viacAMP protein kinase A activity glucuronide secretionactivation of existing TRPV6
Not via VDR but Membrane Associated Rapid Response Steroidbinding (MARRS) protein May also be some effect of calcitriol on TRPV6 synthesis (slow)

Ca2+ dependent FBI may be regulated by calcitriol

Movement across cell--models

1. Calbindin (ferry) protein
1. Induced by calcitriol via VDR 2. probably minor/unlikely 3. See localized, not diffuse Ca after Ca dose

2. Vesicular/lysosomal
1. endocytosis 2. Much evidence

3. Diffusion through the endoplasmic reticulum

1. Vesicles may bud off

The first model

Lysosomal and ER Models

Extrusion at basolateral membrane

1. Not rate-limiting 2. Ca2+ ATPaserequires energy, against concen gradientregulated by calcitriol?? 3. Expression of PMCA is not regulated by calcitriol 4. Exocytosis of vesicles

Functions of Calcitriol related to bone and mineral metabolism

Increase Ca absorption, SI Increase phosphate absorption SI (since it FGF23) Increase renal resorption of Ca (and phosphate) less loss (since it FGF23)
Bone: roles in mineralization (also 24, 25 dihydroxy D) by providing Ca and PO4, differentiation of osteoblasts and remodeling (?)
Differentiation of macrophages to osteoclasts PTH promotes bone resorption

BONE
Cortical
tubular, dense (compact) long bone shaft

Trabecular/cancellous
spiky,spongey Hip, vertebrae, ends of long bones

BONE CELLS
Osteoblasts
embedded in collagen matrix move mineral from ecf to bone surface Become encased in bone--osteocytes

Macrophages differentiate into osteoclasts

Differentiation involves calcitriol Release enzyme to solubilize mineral Increases plasma Ca and PO4 Remodelling of bone, excavation

Calcitriol is needed for osteoclast development

Receptor Activator of NF-KB Ligand); on osteoblast surface; it binds to a receptor on the osteoclast and promotes its differentiation; RANKL by PTH and calcitriol

osteocalcin Expr reg by calcitriol monocyte Osteoblast surface

OPG: osteoprotegerin

Collagenase, HCl

Mineralization
Requires adequate ecf concentrations of Ca and PO4
Ion product (mg/dL) > 40: 10 (Ca) x 4(PO4) = 40 If conc of either ion is too high: calcification of soft tissue (vit D toxicity) If ion product too low: defective mineralization

Phosphorus
As phosphate PO4 3- in diet, in body Organic forms
Phospholipids Nucleic acids Phosphorylated proteins coenzymes: FAD, NAD, CoA, TPP, PLP Inorganic forms Ca3 (PO4)2 hydroxyapatite

Osteoporosis
Decreased bone mass, normal histology Bone pain, easy fracture Diagnosis by bone densitometry
Bone Mineral Density by DXA (dual energy Xray absorptiometry); ultrasound, CT scan Unlike osteomalacia it is not curable by nutritional therapy alone Increased risk with age, menopause, low peak bone mass

Causes of Osteoporosis
Normal decline in bone mass with age
TRPV post-menopause so Ca absorption loss of Ca in urine conversion of vit D to calcitriol vit D synthesis in skin?

Low peak bone mass

Poor Ca and vit D intakes during bone development Low physical activity Genetic predisposition

Bone formation and mineralization

Depends on the balance between osteoblast and osteoclast activity

Therapy for Osteoporosis

Ca supplements have little effect, only cortical bone Hormone therapy risky (CVD, reproductive cancer) Use of drugs that affect estrogen receptors Calcitriol supplements work in elderly women but concern about negative effects

Summary

Genomic Functions of Calcitriol

Nuclear VDR: skin, mammary, gonads Induced by estrogen? Cortisol?(since post weaning)
VDR-RXR dimer and other forms
Each charged with ligand (calcitriol, 9cisRA )

Vitamin D response elements (VDRE) on DNA Regulation of transcription of > 50 genes

Calbindins, osteocalcin, vit D receptors, collagen PTH, FABP, transferrin receptor

osteocalcin
Most abundant bone proetin after collagen Found in bone and in blood Carboxylated form binds to bone mineral (hydroxyapatitea0 Un-carboxylated osteocalcin increases plasma insulin level, proleration of pancreatic cells, and stricter blood glucose control ?role of calcitriol in diabetes?

Vitamin D and Breast Cancer

Many studies show inverse rel between 25 OH D levels in blood and colon or breast cancer activated VDR promotes apoptosis
Mammary cancer cells have 1 hydroxylase and VDR In breast tumors vitD promotes cell differentiation, apoptosis and to reduce the effects of estrogen on proliferation

Type 1 Diabetes mellitus

Incidence of T1DM related to latitude and inversely to hours of sunlight Lack of osteocalcin? Autoimmune disease
Vit D may inhibit production of IL-12 and therefore activation of cytotoxic macrophages and lymphocytes

Suggestive studies that vit D supplements may lower risk

Type 2 Diabetes (circumstantial evidence)

Incidence of T2DM highest in winter (Sweden)

VDR ko mice are more insulin sensitive and resistant to diet-induced obesity
25 hydroxy D levels inversely related to BMI, but calcitriol levels normal in obesity Chronic inflammation inversely related to 25 hydroxy D levels in plasma

Vitamin D Deficiency--Causes
Lack of sun exposure Low dietary intake catabolism: anticonvulsant use (phenobarbital, diphenylhydantoin) Hypoparathyroidism Renal or hepatic disease Mutations in VDR (autosomal recessive)

Vitamin Requirement
Based on vitamins effect on bone, not onother functions
Rickets osteomalacia

Plasma <10 ng/mL but some scientists claim 30 ng/ml should be the normal level
Would need intakes of 3000 IU to achieve that

Rickets
Can be due to defic of Vit D and/or Ca Onset 6-24 mos of ageimpaired mineralization Bone pain Delayed tooth eruption Delayed closure of fontanelles Enlarged epiphyseal plates Bowed legs, knock knee, sabre tibia Rib deformities (respiratory problems) Spinal, Pelvic deformities (childbirth)

Osteomalacia
Onset in older children and adults (once epiphyses have closed Decreased bone mineral Easy fracture Bone pain, muscular weakness Osteopenia (low bone density): increased ratio of non-mineralized to mineralized bone

Vitamin D Toxicity
Dietary supplements NOT sun exposure DRI 5-10 g/d vs UL 50g/d
Some say UL of 4000IU (100g) is safecontroversial since studies done with old assay systems

toxicity defined by hypercalcemia, but other toxic effects may occur at lower levels of D intake than those needed for hypcercalcemia

Vitamin D Toxicity
25 OH D 25 hydroxy vit D competes with calcitriol for VDR, less potent than calcitriol but much higher conc
Ca absorption from SI and bone
Hypercalcemia, calcinosis (CaPO4 deposits in soft tissue), calcinuria. Renal calculi (esp in aged)

PTH in response so less Calcitriol formed Anorexia, vomiting