LESSON 6: LIPIDS AND LIPOPROTEINS

QUESTIONS FOR RESEARCH:

1. Define the following terms: lipid, lipase, atherosclerosis, cholesterol, fatty acid,
hyperlipoproteinemia, triglyceride, glycerol, lipoprotein, lipase, LDL, HDL, lipoprotein, VLDL, and
apolipoprotein.

A lipid is any organic molecule that is insoluble in water. They include fats, waxes, oils,
hormones, and some membrane components that serve as energy storage molecules and
chemical messengers. Lipids, along with proteins and carbohydrates, are important structural
components of living cells.

Lipases are enzymes that degrade triglycerides into free fatty acids and glycerol. Hepatic
lipases, for example, are found in the liver, while hormone-sensitive lipases are found in
adipocytes, lipoprotein lipase is found on the arterial endothelium surface, and pancreatic lipase
is found in the small intestine.

Atherosclerosis is the gradual hardening and constriction of your arteries caused by

cholesterol plaques that form on the arterial lining. It can impair blood flow by clogging your
arteries.

Cholesterol is a waxy, fat-like substance that is found in all cells. The body requires cholesterol
to generate hormones, vitamin D, and digestive chemicals. The body produces all of the
cholesterol it needs. Cholesterol can also be found in animal-derived foods including meat,
cheese, and egg yolks.

Fatty acids are important components of the lipids found in plants, animals, and microbes. A
fatty acid typically consists of a straight chain that has an even number of carbon atoms,
hydrogen atoms running the length of the chain, a carboxyl group (COOH) at one end, and
hydrogen atoms running the length of the chain. It is an acid because of the carboxyl group
(carboxylic acid).

The presence of significant amounts of one or more types of lipoproteins is known as

hyperlipoproteinemia, often referred to as hyperlipidemia or high lipoproteins. A secondary
consequence of underlying medical illnesses or primary genetic diseases may be the cause of
hyperlipoproteinemia. The clinician needs to start looking for main causes after eliminating out
frequent secondary causes of hyperlipoproteinemia.

The most prevalent type of circulating fat is triglycerides. They are made up of three fatty acid
chains connected by the glycerol molecule.

Glycerol is a triol with hydroxy groups substituting for propane at positions 1, 2, and 3. It
functions as a human metabolite, an algal metabolite, a Saccharomyces cerevisiae metabolite,
an Escherichia coli metabolite, a mouse metabolite, a solvent, a detergent, a geroprotector, and
other things.

Any member of the class of molecules known as lipoproteins has both lipid (fat) and protein.
Both soluble and insoluble complexes, such as those found in cell membranes and mammalian
blood plasma, contain them. Because they transport cholesterol through the bloodstream and
lymphatic fluid, lipoproteins in blood plasma have undergone extensive research.

The healthy cholesterol that is helpful for the heart is called high-density lipoprotein. The way

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 that this cholesterol functions is by carrying harmful low-density lipoprotein and very-low-density
lipoprotein to the liver, where they are broken down and eliminated by the body.

Low-density lipoprotein causes serious cardiovascular system damage. It accumulates on

artery walls, stiffening them and bringing on blood clots. The blood pressure rises as the heart
works harder to force blood through the blocked blood arteries.

When levels of very-low-density lipoprotein are too high, your risk of heart disease increases
because it contains even more triglycerides, a kind of fat, than low-density lipoprotein does.
VLDL levels should be between 2 and 30 milligrams per deciliter to be considered healthy.

Apolipoproteins are amphipathic molecules that can interact with both the plasma's aqueous
environment and the lipids in the lipoprotein core. They serve as biochemical keys that provide
lipoprotein particles access to particular places where lipids can be delivered, accepted, or
modified.

2. How are fatty acids, cholesterol, phospholipids and triglyceride metabolized? What are their
functions?

Despite having a varied R group that is a lengthy hydrocarbon chain, all fatty acids share the
same fundamental structure. The chain of carbon molecules is said to be saturated if there are
no double covalent bonds between them. Unsaturated molecules are those that have one or
more double bonds. The chain kinks as a result of a double bond.

Three fatty acids are joined to the glycerol molecule that makes up a triglyceride. In a
condensation reaction, each fatty acid forms an ester link with the glycerol molecule
(esterification). One water molecule is produced by each condensation reaction. In a hydrolysis
procedure, water is injected to break each ester link and release the fatty acids. Water and a
triglyceride are created by joining three fatty acids to glycerol.

A triglyceride's capacity as an energy substrate depends on the following characteristics:

 The hydrophobic tails of the fatty acids (repel water). The glycerol end faces outward,
and the tails of the fatty acids point inward, away from any water, to form droplets in
cells.
 When the hydrocarbon tails are broken down, a lot of energy is released.
 Because they are insoluble in water and do not osmotically draw water into cells, they
have no effect on the water potential of living things. This is advantageous since
triglyceride-rich cells would otherwise quickly enlarge.

A phospholipid is another significant chemical that resembles a triglyceride. This time, a glycerol
molecule is connected to only two fatty acids, and the glycerol molecule also has a phosphate
group. Cell membranes are constructed from phospholipids by forming a bilayer. As is common
knowledge, phosphate groups face outward while fatty acids, which are hydrophobic and repel
water, face inward. To compounds that are dissolved in water, this creates a barrier.

3. What are the lipoproteins and how are they metabolized? What are apolipoproteins?

 Starting in the colon, food lipids are incorporated into chylomicrons to form the exogenous
lipoprotein route. Lipoprotein lipase breaks down the triglycerides carried by chylomicrons in the
circulation, releasing free fatty acids that are then broken down by muscle and adipose tissue
and forming chylomicron remains. Remaining chylomicrons are then absorbed by the liver. The

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 production of VLDL in the liver is the first step in the endogenous lipoprotein pathway.
Lipoprotein lipase breaks down the triglycerides transported by VLDL in muscle and adipose
tissue, releasing free fatty acids and resulting in the formation of IDL. The liver is the main site of
absorption for the LDL receptor, which is where the IDL are primarily absorbed after being
further converted to LDL. The liver and intestines produce nascent HDL, which is the first step in
the reverse cholesterol transport process. The production of mature HDL is facilitated by the
ability of these small HDL particles to take up cholesterol and phospholipids that are effluxed
from cells. ABCA1 mediates this process. ABCG1, SR-B1, or passive diffusion are three ways
that mature HDL might pick up more cholesterol from cells. By interacting with hepatic SR-B1 or
indirectly by transferring the cholesterol to VLDL or LDL, a process made possible by CETP, the
HDL subsequently delivers the cholesterol to the liver. HDL cholesterol efflux from macrophages
is crucial for preventing atherosclerosis from developing.

Apolipoproteins are proteins that bind lipids, or molecules that are soluble in oil, like cholesterol
and fat, to create lipoproteins. Lipids are transported in the blood, cerebrospinal fluid, and lymph
(together with fat-soluble vitamins). Lipoproteins include lipids that are insoluble in water.

4. What other disease state cause increase or decrease in specific lipids and lipoproteins? What
disorders affect apolipoprotiens?

 The treatment of different hormones and endocrine disorders can change plasma lipid levels,
which may raise or lower the risk of atherosclerotic cardiovascular disease. The literature
frequently contains research that report conflicting results, making it difficult to compare them.
These variations could be brought on by variations in the degree of the disease state, variations
in the length of the disease, underlying genetic variations, variations in environmental variables
like diet, the presence of other abnormalities that can alter lipid metabolism such as obesity or
diabetes, and other unrecognized variables that could influence the expression and
manifestation of various endocrine disorders on lipid parameters. LDL and total cholesterol
levels are known to rise in prolactinomas. In contrast to growth hormone therapy, which lowers
total cholesterol and LDL cholesterol while raising Lp(a) levels, growth hormone deficient
patients frequently have higher levels of total cholesterol, LDL cholesterol, and triglycerides and
lower levels of HDL cholesterol. Although ironically comparable to growth hormone insufficiency,
acromegaly is associated with an increase in Lp(a) levels as well as an increase in plasma
triglycerides and a decrease in HDL cholesterol. Total cholesterol, LDL cholesterol, and Lp(a)
levels rise as a result of hypothyroidism, while triglyceride and HDL cholesterol levels remain
stable or rise. The levels of total cholesterol, LDL cholesterol, and Lp(a), as well as HDL
cholesterol, fall with hyperthyroidism, in contrast. The use of glucocorticoids frequently also
raises HDL cholesterol levels, whereas patients with endogenous Cushing's syndrome typically
show an increase in total, LDL, and triglycerides. Although this association is complicated by
obesity and the metabolic syndrome, a major cause of hypogonadism, low testosterone levels in
men may be associated with increased LDL cholesterol and triglyceride levels as well as
decreased HDL cholesterol levels. LDL cholesterol, triglycerides, and Lp(a) rise while HDL
cholesterol falls as a result of androgen deprivation therapy. While high dose androgen therapy
used by sportsmen can significantly lower HDL cholesterol and Lp(a) levels, the effect of
testosterone replacement therapy on plasma lipids and lipoproteins is moderate and
unpredictable. With either no change or a slight decline in HDL cholesterol, the loss of estrogens

(postmenopausal females) is linked to a slight rise in LDL cholesterol. When estrogen is

administered, LDL cholesterol and Lp(a) levels fall while triglycerides and HDL cholesterol levels
rise; however, the dose and method of delivery have an impact on these outcomes (transdermal
has smaller effects). Depending on the androgenicity of the progesterone, concurrent
progesterone therapy may reduce the effect of estrogen on HDL cholesterol and triglyceride
levels while having little to no influence on the decrease in LDL cholesterol brought on by

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 estrogen delivery. Increases in LDL cholesterol, triglycerides, and Lp(a) and decreases in HDL
cholesterol are linked to polycystic ovarian syndrome. An increased risk of atherosclerotic
cardiovascular disease may be brought on by the dyslipidemia that results from prolactinomas,
growth hormone deficit, Cushing's syndrome, male hypogonadism, androgen deprivation
therapy, polycystic ovarian syndrome, and the loss of estrogens.

5. What are the principal laboratory methods of analysis for cholesterol, triglyceride, and the
lipoproteins and apolipoproteins? What interferences might lead to misinterpretation of the
results of these assays? Discuss the principle and procedure of each assay.

Blood samples acquired by venipuncture are typically analyzed for lipoproteins, triglycerides,
and plasma or serum total cholesterol in tubes containing EDTA. Plasma levels have been
found to be about 3% lower than comparable serum levels because of a higher water content.
While triglyceride levels vary greatly from day to day and are highest 1 to 4 hours after meals,
cholesterol levels are generally stable.

Blood should be drawn for triglyceride testing following a 12-hour fast, when chylomicrons have
typically been eliminated from circulation. The best time to take measurements is when the
patients are eating normally and without taking any drugs that can affect their blood lipid levels.
Sampling should not be done during stressful times or within six weeks after a serious illness,
such as an acute myocardial infarction, as these conditions may cause plasma cholesterol
levels to drop and triglyceride levels to rise.

The Abell-Kendall (A-K) method, which includes the Liebermann-Burchardt reaction after
hydrolyzing and extracting cholesterol, is the accepted chemical method for determining plasma
cholesterol levels. In clinical laboratories, entirely automated procedures are frequently used to
analyze samples for plasma cholesterol and triglyceride measurements. It is recommended to
ask how the method employed by the specific laboratory compares with reference values and to
use the same laboratory facility for serial measurements because the results obtained at
commercial laboratories consistently differ from those acquired by research methods. The Lipid
Research Clinics Program, which polled 12,595 Americans aged 20 to 79 in the United States
and Canada, produced the best reference values currently available for North American
populations. Using autoanalyzer systems to which either the ferric chloride-sulfuric acid method
or the Liebermann-Burchardt test was adapted, reference values for plasma total cholesterol
were determined. Triglyceride reference values were determined using a fluorometric technique.
Following the precipitation of apoprotein B-containing lipoproteins in whole plasma (LDL and
VLDL) by heparin-manganese chloride, plasma high-density lipoprotein cholesterol (HDL-C)
measurement in this program was assessed using the same techniques as for plasma total
cholesterol.

Examining a sample of fasting plasma that has been chilled to 4°C is a helpful additional test;
chylomicrons, if present, form a creamy layer at the test tube's top. With a clear infranatant on
top, type 1 hyperlipoproteinemia can be distinguished from type 5 hyperlipoproteinemia with this
test (creamy layer on top with turbid infranatant).

Precipitation, electrophoresis, and ultracentrifugation are methods for separating lipoproteins.

Preparative or density-adjusted ultracentrifugation is methods that can be used to quantify
lipoprotein fractions. This approach is typically used in complex research rather than as a
standard clinical tool. Precipitation and ultracentrifugation are used consecutively in the
quantitative reference method to identify each lipoprotein component.

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 Advanced laboratories use radioimmunoassays (for apoprotein AI and apoprotein B) and
isoelectric focussing to quantitatively determine apoproteins (for apoproteins C and E).

Other specialized tests include the intravenous heparin test for assessment of postheparin
lipolytic activity (this test measures the preheparin and postheparin liberation of lipoprotein
lipase and is primarily used to confirm congenital lipoprotein lipase deficiency), and the
assessment of LDL receptor status on cultured skin fibroblasts or isolated lymphocytes.

REFERENCES/SOURCES:

Indicate the learning materials you used in accomplishing this self-directed activity.

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Facts. Encyclopedia Britannica. https://www.britannica.com/science/lipid
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https://www.ncbi.nlm.nih.gov/books/NBK537346/
 Atherosclerosis. (2007, October 26). WebMD. https://www.webmd.com/heart-disease/what-is-
atherosclerosis
 Cholesterol. (n.d.). MedlinePlus. Retrieved October 29, 2022, from
https://medlineplus.gov/cholesterol.html
 The Editors of Encyclopaedia Britannica. (2022, September 14). fatty acid | Definition, Structure,
Functions, Properties, & Examples. Encyclopedia Britannica.
https://www.britannica.com/science/fatty-acid
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 Harvard Health. (2020, March 1). Understanding triglycerides.
https://www.health.harvard.edu/heart-health/understanding-triglycerides
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 Anne, M. (2018, December 14). Definition of HDL, LDL & VLDL. Healthy Eating | SF Gate.
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 Fatty Acids, Triglycerides and Phospholipids – Biological Molecules Ep 4. (2021, June 26). Zoë
Huggett Tutorials. https://zhtutorials.com/2020/10/17/fatty-acids-triglycerides-phospholipids/
 Feingold, K., Anawalt, B., & Boyce, A. (2021, January 19). Introduction to Lipids and Lipoproteins.
National Library of Medicine. https://pubmed.ncbi.nlm.nih.gov/26247089/
 Feingold, K. R., Brenton, E. A., & Grunfeld, C. (2020, March 9). The Effect of Endocrine Disorders
on Lipids and Lipoproteins. National Library of Medicine.
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