Lipids & Lipoprotein

Part 2
 Lipoprotein Pathways
• Lipid absorption, exogenous, and
endogenous pathways all used apo B-
containing lipoproteins
– Transport dietary and hepatic derived lipid to
peripheral cells
– Transport energy to cells but also cholesterol
– If too much – cholesterol accumulates
• Peripheral cells cannot catabolize
• Hydrophobic cannot diffuse away

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Lipid Pathways

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 Lipid Absorption
• Average person takes in 60-130 grams of
fat per day
– Mostly triglycerides
– Pancreatic lipase cleaves FA
– Triglycerides → mono or diglycerides
– Cholesteryl esters → free cholesterol
– These amphipathic molecules aggregate in
intestine with bile acids into micelles

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 Lipid Absorption
• Micelles contact intestinal villi and are
absorbed – passive and active transport
– Smaller FA (<10 carbons) absorbed directly
into portal circulation
– Larger fragments converted back into
triglycerides & cholesteryl ester
– Packaged chylomicrons with apo B-48
• > 90% absorption effectiveness for
triglycerides but only 50% for cholesterol

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 Exogenous Pathway
• Newly synthesized chylomicrons secreted
into lymphatic ducts → thoracic duct →
blood
• Interact with lipoprotein lipase
– Hydrolyzes triglycerides
– Free FA and triglycerides = energy for cells
– Excess – reesterified for storage
– Mobilized with hormone sensitive lipase

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 Exogenous Pathway
• Lipid and apolipoproteins transferred onto
HDL leaving chylomicron remnants
• Chylo-remnants taken up by liver via apo
E and liver receptor
• Liver breaks down further to FA,
triglycerides, cholesterol, and amino acids

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 Endogenous Pathway
• Triglyerides in liver packaged into VLDL
(from dietary recirculation from adipose
tissue)
• Only small fraction synthesized de novo
• VLDL lipolysis similar to chylomicrons
– Loss of core lipids - Dissociation of particle
– Transfer to other particles
– VLDL remnants - half are converted to LDL,
half taken up by liver

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 Endogenous Pathway
• LDL primary transport of exogenous
cholesterol to cells
– LDL receptors very efficient
– Endocytosed & degraded
– Triglycerides for energy or stored
– Free cholesterol membrane biosynthesis
• Excess cholesterol → cholesteryl ester & stored

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 Endogenous Pathway
• Biosynthesis enzymes for cholesterol
downregulated when excess cholesterol in
cells
– Target for cholesterol-statin drugs
• Abnormal LDL receptor function →
hypercholesterolemia → premature
atherosclerosis

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 Reverse Pathway
• HDL is thought to remove excess cellular
cholesterol by 2 mechanisms
1. Aqueous diffusion pathway
• Small amount of cholesterol diffuses away from
cells
• Converted to cholesteryl ester by LCAT (lecithin
cholestrol acyltransferase)
• About ½ transferred to LDL and taken back to
liver
– By Cholesterol ester transfer protein (CETP)
– Excreted into bile or converted to bile acids

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 Reverse Pathway
• HDL is thought to remove excess cellular
cholesterol by 2 mechanisms
2. ABCA1 transporter pathway
• ATP binding transporter group
• Pumps ligands across cell membrane

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• Diseases associated with abnormal lipid
concentrations are referred to as
dyslipidemias
– Genetic abnormality
– Environmental /lifestyle imbalances
– Secondary

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 Arteriosclerosis
• Deposition of esterified lipids in
artery walls = fatty streaks
– Small damage to walls - macrophages
& platelets move in to repair
– LDL brings cholesterol for new cell
membranes
– LDL can also be taken up by
macrophages = foam cells
– Accumulation of foam cells deposit in
artery walls = fatty streaks

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 Arteriosclerosis
• Additional injury
(inflammation) leads to
more deposition = plaque
formation
– Plaques narrow the artery
opening which assists
additional deposits and
narrowing

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 Arteriosclerosis
• Increased LDL and decreased HDL
associated with increased plaque
formation
– 1% decrease in LDL drops CHD risk 2%
– Below 100 mg/dl for established patients for
stability
• Some are due to genetic factor
– Too much synthesized or too little removed

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 Dyslipidemias
• Disease states are generally caused by
malfunction in synthesis, transport, or
catabolism of lipoproteins
• Dyslipidemia
– hyolipoproteinemia
– Hyperlipoproteinemia
• Hypercholesterolemia
• Hypertriglyceridemia
• Combined hyperlipoproteinemia

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 Hypercholesterolemia

• Familial hypercholesterolemia (FH)

– Homozygotes rare 1/million
• Total cholesterol 800-1000 mg/dl
• Heart attack as early as teenage years
– Heterozygotes cholesterols 300-600
mg/dl
• Heart attacks 20-50 years

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 Hypercholesterolemia
• Familial hypercholesterolemia (FH)
– Primarily LDL elevations
– Synthesis is normal but decrease or lack LDL
receptors
– Therefore LDL builds-up in serum
– Since cells cannot acquire from LDL increase
internal synthesis

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 Hypertriglyceridemia
• Triglycerides
– Borderline = 150-200 mg/ dl
– High 200-500 mg/dl
– Very High > 500 mg/dl
• Familial hypertriglyceridemia
– genetic
• Secondary hypertriglyceridemia
– hormonal imbalances
– Imbalance between synthesis and clearance of VLDL

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 Hypertriglyceridemia
• Generally caused by deficiency of LPL or
LPL cofactor.
• LPL hydrolyzes triglycerides in
chylomicrons and VLDL
• Deficiency prevents processing and
clearing
• Elevated even with fasting

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 Hypolipoproteinemia
• Deficiency of lipoproteins
• Hypo alpha or hypobetalipoproteinemias
• Hypobetalipoproteinemia
– Associated with low LDL
– Not associated with CHD

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 Hypolipoproteinemia
• Hypoalphalipoproteinemia
– Several genetic defects
– Isolated decreased in circulating HDL
(<40mg/dl) without hypertriglyceridemia
– Premature CHD
– Tangier Disease may have HDL 1-2 mg/dl

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 Hypolipoproteinemia
• Hypoalphalipoproteinemia
• Acute transient hypoalphalipoproteinemia
– Severe physiologic stress
– Infection, surgery, generalized illness
– Therefore samples taken during
hospitalization interpret with caution
– Return to normal after recovery

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Cholesterol Enzymatic Method

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Triglyceride measurement

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 Lipoprotein Methods
• Must separate fractions
– Electrophoresis – agarose or polyacrylamide
– Chromatographic
– Precipitation
– Ultracentrifugation
– Immunochemical

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 HDL Methods
• CDC reference method
• Three step process
– Ultracentrifugation to remove VLDL
– Heparin manganese precipitation to remove LDL
activity
– Then analysis

• Two-step method with precipitation of non-HDL

fractions (apo-B) then analysis on supernatant
– Enzymatic cholesterol method on remaining

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 LDL Methods
• Friedewald calculation
– Determine total cholesterol, total triglycerides
and HDL as before
– VLDL is estimated as triglycerides/5
• 400 mg/dl is the upper limit
• LDL = Total Cholesterol – HDL – Trig/5

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 Apolipoprotein Methods
• Primarily a research method or specialty
cardiac service center
– Usually done by Immunoassay –
• turbidimetric
• ELISA, RIA

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