Ageing

SBM 10/8/6

by Dr. P. H.P. Fernando

Cellular Contribution to Ageing

Relatively long lifespan of humans low level of free radical production  low level of fatty acid unsaturation  high level of DNA repair enzymes


Free radical theory of ageing

Cause damage to

DNA Polyunsaturated fatty acids Proteins and amino acids Membrane structures

Reactive Oxygen Species (ROS) ROS) hydroxyl radical is the strongest attacker bases in nucleic acids amino acid side chains in proteins double-bonds in unsaturated fatty acids doubleoxidative stress. stress. Reactive Nitrogen Species (RNS) RNS) Peroxynitrite mostly damage endothelial cells

Protection from Oxidative Stress

Antioxidants Glutathione Vitamin C, Vitamin E, Vitamin A Carotenoids, Selenium Uric Acid Enzymes Superoxide dismutase (Cu 2+ / Zn 2+ ) Glutathione peroxidase Peroxidase Catalase

Mitochondria and Aging

Increasing insulin levels associated with aging stimulates nitric oxide synthetase resulting in peroxinitrile Lipid peroxidation of the inner mitochondrial membrane Increase proton leak independent of uncoupling protein Degrade function of respiratory enzymes Inactivate mitochondrial superoxide dismutase

Aging
decreased oxidative phosphorylation coupling efficiency increased superoxide production

"Viscious cycle"
free radical damage to mitochondrial DNA mitochondria that produce more superoxide

Lysosomes & mitochondria

Most damaged mitochondria are consumed by lysosomes more defective mitochondria (which produce less ATP as well as more superoxide) remain to reproduce themselves Efficiency of lysosomes to consume malfunctioning mitochondria declines with age, resulting in more mitochondria producing higher levels of superoxide

Mitochondria of older organisms

fewer in number larger in size less efficient produce less energy and more superoxide Less CoQ protective against oxidative stress

Glycation in ageing
nonnon-enzymatic glycosylation - Maillard reaction reducing sugars become attached to proteins without the assistance of an enzyme.
-

Amadori product (a ketoamine) Both glycation and Amadori product formation are completely reversible reactions. Advanced Glycation End-products (AGEs) formed by EndAGEs) oxidation of Amadori products is irreversible.

Glycation in ageing

Effects of AGEs
AGEs in tissues increase the rate of free radical production to 50-times the rate of free-radical 50freeproduction by unglycated proteins AGEs In LDL - accelerates oxidation and atherosclerosis. In vessel collagen also atherosclerosis kidney failure Cataracts

Glycation potential
Glucose x 1 Galactose x 5 Fructose x 8 Ribose x 100 Deoxyribose x 200 (Glucose assumes cyclic conformation readily) Some aldehyde products of lipid peroxidation

Protein damage and maintenance in ageing

Glycation , Free Radical Damage, Racemation Oxidation Cysteine & methionine most vulnerable 30 50% of proteins oxidized in old animals Protein cross linking by the formation of dityrosine bridges Deamination (spontaneous) of asparagine (400 times greater than for gln and glutamine on proteins, especially when a glycine is in the adjacent carboxyl position.

Proteolytic enzymes
Cellular proteins are continually being degraded (hydrolyzed) within cells by proteolytic enzymes, for regulation of cellular processes, and for "quality control" of proteins

Proteolytic enzymes
The four major classes of cellular proteolytic enzymes are caspases calpains cathepsins proteasomes Proteasome activity declines with age.

HeatHeat-shock proteins HSPs

Cellular protection from thermal and other stresses Increased by a transient elevation of temperatures that could ordinarily kill a cell. The magnitude of induction of heat-shock proteins heatis significantly reduced with aging Incidence of heat stroke among those 65 years of age or older is ten times that of younger persons.

DNA damage and DNA repair

"Wear & tare" of DNA can take two forms: mutation DNA damage DNA damage tends to interfere with gene expression by preventing transcription of RNA from DNA DNA damage rather than DNA mutation is posited as a cause of aging.

Types of DNA damage

SingleSingle-strand break N7 MethylGuanine Depurination O6 MethylGuanine Oxidized DNA Depyrimidation Cytosine deamination DoubleDouble-strand breaks Interstrand cross-links cross-

rrr correlated with Maximum Life Span (MLS)

Mitochondrial DNA
Healthy adults 6580 years of age have about 25% higher skeletal muscle mtDNA 8oxodG than adults 2035 Even without oxidative damage mtDNA mutations and deletions promote apoptosis, leading to tissue degeneration and aging

Telomeres and ageing

At the ends of each chromosome is a long non-functional strand of DNA called a nontelomere. telomere. Telomeres consist of the six-base repeating sixsequence TTAGGG With each cell division, some of the telomere is lost. Therefore, the number of times that most dividing cells can divide is limited.

Telomerase
Germ cells, stem cells and "immortalized" cancer cells contain telomerase that replaces lost telomeres, Telomerase is a reverse transcriptase, transcriptase,

Accumulation of toxins and chemical garbage

Lipofuscin (age pigment), which can accumulate in large quantities in nonnondividing cells. Lipofuscin is regarded as a product of lysosomes As production of lysosome enzymes decline with age -- and as lysosomes engulf increasingly cross-linked proteins & lipids crossthat are resistant to enzyme degradation --

Lipofuscin continued
dysfunctional lysosomes (bloated with indigestible contents) accumulate in cells as lipofuscin granules. Lipofuscin granules are characterized by a single membrane envelope, enclosing yellowishyellowish-brown material that can autofluorescence.

Apoptosis
programmed cell suicide genetically controlled cell death causes cells to shrink and be eliminated without the tissue truamas associated with inflammation that accompanies uncontrolled cell death (necrosis). necrosis). Benefits eliminating defective cells and protecting from cancer be associated with harmful conditions, as in atherosclerosis and neurogenerative disease.

Apoptosis
caspases -- Cysteine Aspartase ProteASES) roteASES) Apoptosis initiated by an extracellular signal (Fas receptor) activates caspase 8, whereas apoptosis due to intracellular damage or distress activates caspase 9.

Senescent cells
cells that no longer proliferate or divide in response to growth factors or mitogens can function like pre-senescent cells prebut display a number of distinctive characteristics.

Senescent cells - distinctive characteristics. characteristics.

increased free radical production increased oxidative damage increased glycation damage reduced heat shock protein expression

Senescent cells
senescent cells are "old ? genetic down-regulation of proteasome downactivity Senescent cells are resistant to apoptosis. decline in apoptotic protein function ?

Caloric restriction with adequate nutrition (CRAN)

adequate nutrition - vitamins, minerals, essential amino acids and essential fatty acids in adequate quantity Rats, mice and hamsters experience maximum lifespan extension from a diet which contains 4060% of the calories Mean lifespan is increased 65% Maximum lifespan is increased 50% when CRAN is begun just before puberty

Mechanism of CRAN ?
maturity, thymus shrinkage, DNA-repair decline DNAand tumor formation delayed. complete oxidation of fatty acids, with fewer ketones in the blood cell membranes having less cholesterol & saturated fatty acids. Collagen cross-linking occurs more slowly crossblood glucose levels reduced about 15% below controls. Reduction of body fat is associated with reduced insulin resistance due to reduced levels of the proinflammatory cytokine

Thank you