Ethanol Metabolism (6 orang)

Ethanol is a small two carbon alcohol that, due to its small size and alcoholic hydroxyl group
is soluble in both aqueous and lipid environments. This allows ethanol to freely pass from
bodily fluids into cells. Since the portal circulation from the small intestines passes first
through the liver, the bulk of ingested alcohol is metabolized in the liver. The process of
ethanol oxidation involves at least three distinct enzymatic pathways.

The most significant pathway, responsible for the bulk of ethanol metabolism, is that
initiated by alcohol dehydrogenase, ADH. As outlined below, humans express several ADH
genes with the class I members being responsible for hepatic ethanol metabolism. The ADH
enzymes are NAD+-requiring and they are expressed at high concentrations in hepatocytes.
Animal cells (primarily hepatocytes) contain cytosolic ADH which oxidizes ethanol to
acetaldehyde. Acetaldehyde then enters the mitochondria where it is oxidized to acetate by
mitochondrial aldehyde dehydrogenase (ALDH). A cytosolic ALDH exists but is responsible
for only a minor amount of acetaldehyde oxidation.

The second major pathway for ethanol metabolism is the microsomal ethanol oxidizing
system (MEOS) which involves the cytochrome P450 enzyme CYP2E1 and requires NADPH
instead of NAD+ as for ADH. The MEOS pathway is induced in individuals who chronically
consume alcohol.

The third pathway involves a non-oxidative pathway catalyzed by fatty acid ethyl ester
(FAEE) synthase. This latter pathway results in the formation of fatty acid ethyl esters and
takes place primarily in the liver and pancreas, both of which are highly susceptible to the
toxic effects of alcohol.

Oxidation of ethanol can also occur in peroxisomes via the activity of catalase. However, this
oxidation pathway requires the presence of a hydrogen peroxide (H2O2) generating system
and as such plays no major role in alcohol metabolism under normal physiological
conditions.
Acetate from EtOH Metabolism and Fatty Liver
Under normal conditions the levels of acetate in human serum is < 0.2mM. Therefore, the
roles of acetate metabolism in mammals under normal physiological conditions remain to
be established. Normal physiological sources of acetate include bacterial fermentation in
the colon which increases significantly when consuming a high fiber diet. This intestinal
acetate enters the portal circulation and is taken up by the liver where it is converted to
acetyl-CoA. Intracellular generation of acetate is the consequence of the ubiquitously
expressed cytosolic enzyme acetyl-CoA hydrolase. The acetate can then be salvaged by
reactivation to acetyl-CoA. In the nervous system, the neurotransmitter acetylcholine (ACh)
is degraded to acetate by acetylcholinesterase. In order to replenish the pool of ACh the
acetate must be reactivated to acetyl-CoA so that it can participate in the choline
acetyltransferase catalyzed reaction.
Acetate is also generated within the nucleus of all cells via the action of histone deacetylases
(HDAC). As with the other sources of acetate, this nuclear acetate must be reactivated
before it can be oxidized or reused.

Under conditions of prolonged starvation and in type 1 diabetes, the endogenous pathways
of acetate production are the main sources for serum acetate. Following the consumption of
ethanol, acetate levels can be elevated by as much as 20–fold.
Acetate, from whatever source, is converted to acetyl-CoA by ATP-dependent acetyl-CoA
synthetases (AceCS: encoded by acyl-CoA synthetase short-chain family member genes:
ACSS) by the following reaction:
ATP + acetate + CoA ←→ AMP + PPi + acetyl-CoA
Humans express cytosolic and mitochondrial acetyl-CoA synthetases, encoded by the ACSS2
and ACSS1 genes, respectively. Expression of ACSS2 is under the control of SREBP-1c which
is itself transcriptionally regulated by PGC-1α. Interestingly, mammalian AceCS are regulated
by reversible acetylation catalyzed by the sirtuins, SIRT1 and SIRT3, and the activity of PGC-
1α is also regulated by SIRT1 activity. Cytosolic ACSS2 is a target of cytoplasmic SIRT1
whereas, mitochondrial ACSS1 is a target of mitochondrial SIRT3.
The primary causes of fatty liver syndrome (hepatic steatosis), induced by excess alcohol
consumption are the altered NADH/NAD+ levels that in turn inhibits gluconeogenesis,
inhibits fatty acid oxidation and inhibits the activity of the TCA cycle (see below for details).
Each of these inhibited pathways results in the diversion of acetyl-CoA into de novo fatty
acid synthesis. Ethanol has also been shown to activate SREBP-1c which results in the
transcriptional activation of numerous genes involved in lipogenesis. However, given that
large amounts of acetate are generated via ethanol metabolism in the liver, this can be a
significant contributor to the overall pool of acetyl-CoA utilized as the precursor for fatty
acid and cholesterol biosynthesis.

Enzymes of Ethanol Metabolism Alcohol Dehydrogenases

In humans there are multiple isoforms of ADH encoded for by seven different ADH genes. All
seven ADH genes reside within a 365kbp region on chromosome 4q23. All human ADH
genes are members of a large family of enzymes known as the medium-chain
dehydrogenase/reductase (MDR) superfamily. Within the human genome there are at least
25 members of the MDR family of genes. Functional ADH exists as either a homo- or a
heterodimer and the active enzymes are divided into five distinct classes denoted I–V. It
should be pointed out that humans evolved to express multiple ADH genes and isoforms,
not for metabolism of ethanol, but to metabolize naturally occurring alcohols found in foods
as well as those produced by intestinal bacteria. As an example, one form of ADH (encoded
by the ADH7 gene) is responsible for the metabolism of not only ethanol but also retinol to
retinaldehyde which is the form of vitamin A necessary for vision.

The hepatic forms of ADH are derived from the protein subunits encoded by the class I
genes:
ADH1A (also known as ADH1), ADH1B (also known as ADH2), and ADH1C (also known as
ADH3). The α, β, and γ subunits encoded by these three genes, respectively, can form homo-
and heterodimers as indicated above. These ADH isoforms account for the vast majority of
ethanol oxidation in the liver. As the concentration of ethanol increases in the liver the π-
ADH isoform, encoded by the ADH4 gene, contributes significantly to overall hepatic ethanol
oxidation. The ADH5 gene is ubiquitously expressed and the encoded protein functions in
the oxidation of long-chain primary alcohols and as a formaldehyde dehydrogenase. The
ADH5 encoded protein exhibits little, if any, ethanol oxidizing activity as can be seen by the
extremely high Km indicated in the Table. The ADH6 encoded enzyme has not been
characterized so little is known regarding its substrate(s) and activity. As indicated above the
ADH7 encoded enzyme oxidizes both ethanol and retinol.

Aldehida dehidrogenase

Ada dua gen aldehida dehidrogenase (ALDH) utama pada manusia yang menyandikan enzim
yang bertanggung jawab atas oksidasi asetaldehida yang dihasilkan selama oksidasi etanol.
Gen-gen ini diidentifikasi sebagai ALDH1A1 dan ALDH2, mereka masing-masing
menyandikan enzim ALDH1 dan ALDH2. Gen ALDH1A1 terletak pada kromosom 9q21.13
dan terdiri dari 13 ekson yang mengkode protein asam amino 501. Gen ALDH2 ditemukan
pada kromosom 12q24.12 dan terdiri dari 13 ekson yang menghasilkan dua mRNA yang
disambung secara alternatif yang mengkode protein dari asam amino 517 (isoform 1) dan
asam amino 470 (isoform 2). Protein ALDH1 adalah enzim sitosol sedangkan protein
ALDH2 berada di mitokondria.

Sebagian besar oksidasi asetaldehid terjadi di mitokondria melalui ALDH2. Namun, beberapa
oksidasi akan terjadi dalam sitosol melalui ALDH1 sebagai alat untuk membantu mengontrol
kadar asetaldehida secara keseluruhan. Fakta terakhir ini paling jelas pada individu dengan
alel ALDH2 yang menunjukkan kapasitas pengoksidasi asetaldehida yang rendah hingga
tidak ada. Beberapa polimorfisme ALDH2 diketahui ada di berbagai populasi. Memang,
variasi gen yang paling banyak dipelajari dalam enzim metabolisme alkohol adalah yang ada
pada gen ALDH2. Alel ALDH2 * 2 menyimpan residu Lys (K) pada posisi 487 daripada
residu Glu (D) yang normal. Alel ini mengkode enzim ALDH2 yang hampir tidak aktif. Dari
signifikansi biokimia dan fisiologis tertentu adalah fakta bahwa alel ALDH2 * 2 bertindak
dalam cara yang hampir dominan sehingga bahkan heterozigot hampir tidak memiliki
aktivitas ALDH2 yang terdeteksi. Alel ALDH2 * 2 lazim pada orang-orang keturunan Cina,
Jepang, dan Korea tetapi pada dasarnya tidak ada pada orang keturunan Afrika atau Eropa.
Alel ALDH2 khusus ini bertanggung jawab untuk kemudahan yang menyebabkan banyak
orang Timur menjadi mabuk oleh konsumsi alkohol dan fakta ini disebabkan oleh
berkurangnya tingkat metabolisme etanol. Selain itu, karena kadar asetaldehida dalam darah
orang-orang ini meningkat dengan cepat setelah konsumsi alkohol, hal itu mengarah pada
reaksi yang sangat merugikan pada senyawa ini yang meliputi pembilasan parah, mual, dan
takikardia

Acute Effects of Ethanol Metabolism

The primary acute effects of ethanol consumption are the result of the altered NADH/NAD+
ratio that is the consequence of both the ADH and ALDH catalyzed reaction. Acute effects
resulting from ethanol metabolism are also due to the fact that acetaldehyde forms adducts
with proteins, nucleic acids and other compounds resulting in impaired activity of the
affected compounds. Additional acute consequences of ethanol metabolism include oxygen
deficits (i.e., hypoxia) in the liver and the formation of highly reactive oxygen-containing
molecules (i.e., reactive oxygen species, ROS) that can damage other cell components.

As indicated above, both the ADH and ALDH catalyzed oxidation reactions lead to the
concomitant reduction of NAD+ to NADH. The majority of the aberrant metabolic effects of
ethanol intoxication stem from the actions of ADH and ALDH and the resultant cellular
imbalance in the NADH/NAD+ ratio. The NADH produced in the cytosol by ADH must be
reduced back to NAD+ via either the malate-aspartate shuttle or the glycerol-phosphate
shuttle. Thus, the ability of an individual to metabolize ethanol is dependent upon the
capacity of hepatocytes to carry out either of these two shuttles, which in turn is affected by
the rate of the TCA cycle in the mitochondria. The rate of flux through the TCA cycle is itself
being negatively affected by the NADH produced by the ADH and ALDH reactions. The
reduction in NAD+ impairs the flux of glucose through glycolysis at the glyceraldehyde-
3phosphate dehydrogenase reaction, thereby limiting energy production. Additionally, there
is an increased rate of hepatic lactate production due to the effect of increased NADH on
direction of the hepatic lactate dehydrogenase (LDH) reaction. This reversal of the LDH
reaction in hepatocytes diverts pyruvate from gluconeogenesis leading to a reduction in the
capacity of the liver to deliver glucose to the blood.

In addition to the effects on biochemical reactions just discussed, the NADH/NAD+ ratio, and
as a consequence the redox state of the cell, is also dramatically altered as a result of
ethanol metabolism. Alterations in cellular redox state are known to affect the level of
expression of certain genes. To appreciate this fact one can look to research done on
calorie-restriction diets. Research on reduced food intake (caloric restriction) has shown
that NAD+ levels may act as a sensor that regulates the activity of certain genes. Activation
of those genes, in turn, has been shown to be related to extended lifespans in a wide variety
of organisms. In addition, the NAD+-regulated gene networks have been shown to reduce
the incidence of age-related diseases, such as diabetes, cancer, immune deficiencies, and
cardiovascular disorders. Therefore, alterations in the NADH/NAD+ ratio (in particular
reductions in NAD+) resulting from ethanol metabolism may result is negatively altered
expression of gene networks that promote healthy cells.

Chronic Effects of Ethanol Metabolism

In addition to the negative effects of the altered NADH/NAD+ ratio on hepatic
gluconeogenesis, fatty acid oxidation is also reduced as this process requires NAD+ as a
cofactor. Concomitant with reduced fatty acid oxidation is enhanced fatty acid synthesis and
increased triglyceride production by the liver. In the mitochondria, the production of acetate
from acetaldehyde leads to increased levels of acetyl-CoA. Since the increased generation of
NADH also reduces the activity of the TCA cycle, the acetyl-CoA is diverted to fatty acid
synthesis. The reduction in cytosolic NAD+ leads to reduced activity of glycerol-3-phosphate
dehydrogenase (in the glycerol 3-phosphate to DHAP direction) resulting in increased levels
of glycerol 3-phosphate which is the backbone for the synthesis of the triglycerides. Both of
these two events lead to fatty acid deposition in the liver leading to fatty liver syndrome and
excessive levels of lipids in the blood, referred to as hyperlipidemia.
Because ethanol metabolism by ADH and ALDH occurs essentially only in the liver, any of the
adverse effects described above that are associated with ethanol metabolism by these
enzymes, and the associated ROS production, primarily affect that organ. In contrast,
CYP2E1 is found in many tissues in addition to the liver, including the brain, heart, lungs,
neutrophils, and macrophages. Accordingly, metabolic consequences of CYP2E1-mediated
ethanol oxidation will affect numerous tissues. The harmful effects associated with CYP2E1-
mediated ethanol metabolism are primarily related to the production of ROS, mainly
superoxide and hydroxyl radicals. In the liver, the oxidative stress resulting from CYP2E1-
mediated ethanol metabolism plays an important role in alcohol-related development of
liver cancer.
Chronic ethanol consumption and alcohol metabolism also negatively affects several other
metabolic pathways, thereby contributing to the spectrum of metabolic disorders frequently
found in alcoholics. These disorders include fatty liver syndromes such as NAFLD and NASH,
hyperlipidemia, lactic acidosis, ketoacidosis, and hyperuricemia. The first stage of liver
damage following chronic alcohol consumption is the appearance of fatty liver, which is
followed by inflammation, apoptosis, fibrosis, and finally cirrhosis.