Slide 1

2001 By Default!

Alcoholic Cardiomyopathy

By, Atman Shah shahatma@msu.edu

A Free sample background from www.pptbackgrounds.fsnet.co.uk

Slide 2

2001 By Default!

Introduction
1. Understanding alcohol metabolism in the human body 2. Research paper studying the effects of excess metabolic byproducts on the heart

A Free sample background from www.pptbackgrounds.fsnet.co.uk

Slide 3

2001 By Default!

Alcohol Metabolism-1

Ethanol is absorbed in the stomach and the intestine and enters the blood.

In the liver and heart, an enzyme called alcohol dehydrogenase (ADH) converts alcohol to acetaldehyde. Acetaldehyde is approximately 30 times more toxic than alcohol, acetaldehyde is a major cause of alcohol-associated side effects. (Drunkenness, hangovers)

A Free sample background from www.pptbackgrounds.fsnet.co.uk

Slide 4

2001 By Default!

Alcohol Metabolism-2
Acetaldehyde is rapidly converted to acetate by other enzymes. Acetate (acetic acid) is is eventually metabolized to carbon dioxide and water.

A Free sample background from www.pptbackgrounds.fsnet.co.uk

Slide 5

2001 By Default!

Research Article

Published in: Journal of Pharmacology and Experimental Therapeutics Date: August 3, 1999
A Free sample background from www.pptbackgrounds.fsnet.co.uk

Slide 6

2001 By Default!

Purpose of This Study:

To show that acetaldehyde overproduction causes Alcoholic Cardiomyopathy (ACM)
The acetaldehyde overproduction is artificially produced by altering genes (transgenic) in mice.
A Free sample background from www.pptbackgrounds.fsnet.co.uk

Slide 7

2001 By Default!

Methods and Results:

1.
2. 3.

Develop transgenic mice

Analysis of transgene expression Chronic treatment of mice with ethanol Study the effects on the heart

4.

A Free sample background from www.pptbackgrounds.fsnet.co.uk

Slide 8

2001 By Default!

Develop Transgenic Mice

The genes responsible for transcribing Alcohol Dehydrogenase (ADH) are altered to produce more ADH The MyADH transgene was produced by replacement of the catalase coding sequence in the transgene MyCAT with the coding sequence for rat ADH

(Alcohol is converted into acetaldehyde by ADH)

A Free sample background from www.pptbackgrounds.fsnet.co.uk

Slide 9

2001 By Default!

Analysis of Transgene Expression

FVB are normal (non-transgenic) mice Line 239 and 258 both show high enzyme activity Line 239 is selected for the study since it also shows a better color coat, useful for identification.

Figure 1: Shows expression of the transgene compared to normal mice. Level of expression is measured by enzyme activity.
A Free sample background from www.pptbackgrounds.fsnet.co.uk

Slide 10

2001 By Default!

Analysis (Cont)

Figure 2: Shows rRNA isolated from various tissues probed with a fragment specific to the transgene product. Multiple Northern blots of different transgenic tissue identified the expression of the MyADH transgene ONLY in the heart.
A Free sample background from www.pptbackgrounds.fsnet.co.uk

Slide 11

2001 By Default!

Will These Mice Really Work in the Study?

I.P. injection of 3g/kg alcohol Amount of acetaldehyde in the mice was detected using gas chromatography.

Figure 3: Shows that transgenic mice have 4 times more acetaldehyde than control mice.

A Free sample background from www.pptbackgrounds.fsnet.co.uk

Slide 12

2001 By Default!

Chronic Treatment of Mice With Ethanol

The animals were initiated on a 1% alcohol (by volume) liquid diet for 10 weeks.

The quantity of alcohol was gradually increased to 4%.

After 10 weeks the mice were sacrificed and their hearts were studied.

A Free sample background from www.pptbackgrounds.fsnet.co.uk

Slide 13

2001 By Default!

Effects on the Heart

Increased expression of:
Alpha-skeletal actin (SkActin) and Atrial Natriuretic factor (ANF)

(Provide a sensitive indicator of cardiac damage in cardiomyopathies)

A Free sample background from www.pptbackgrounds.fsnet.co.uk

Slide 14

2001 By Default!

Indicators of Cardiac Damage (1)

Figure 4: Shows that levels of mRNA of both SkActin and ANF were significantly increased in alcohol treated mice.

A Free sample background from www.pptbackgrounds.fsnet.co.uk

Slide 15

2001 By Default!

Indicators of Cardiac Damage (2)

After 5-months on an alcohol diet mouse hearts had enlarged. There was a more dramatic cardiac enlargement in transgenic mice. Heart-to-Body ratio increased to 18% in control mice and 80% in transgenic mice.

A Free sample background from www.pptbackgrounds.fsnet.co.uk

Slide 16

2001 By Default!

Indicators of Cardiac Damage (3)

Electron microscopy revealed morphological changes in the myocardium of alcohol-treated mice.

Normal mice that were treated showed mild effects such as sarcoplasmic edema. Transgenic mice showed more global effects such as:

Loss of sarcoplasmic reticulum 2. Degenerated and/or smaller mitochondria 3. Disorganized cristae in mitochondria
1.
A Free sample background from www.pptbackgrounds.fsnet.co.uk

Slide 17

2001 By Default!

Electron Microscopy

A and C are not treated with alcohol (control) B and D are treated with alcohol C and D are taken from transgenic mice

A Free sample background from www.pptbackgrounds.fsnet.co.uk

Slide 18

2001 By Default!

Indicators of Cardiac Damage (4)

A pressure transducer was used to measure contractility.

Figure 7: 18-weeks of alcohol diet reduced the contractility in both control and transgenic mice.
A Free sample background from www.pptbackgrounds.fsnet.co.uk

Slide 19

2001 By Default!

Conclusion
Increased acetaldehyde exposure to the myocardium produced alcoholic cardiomyopathy in mice.
Future research:
Trying to produce transgenic mice that have excess acetaldehye dehydrogenase, to protect the heart from the damaging effects of acetaldehyde.

A Free sample background from www.pptbackgrounds.fsnet.co.uk