CELLULAR INJURY

APOPTOSIS AND
NECROSIS
• General Biochemical Mechanisms
Particularly vulnerable are glycolysis, the citric acid cycle and oxidative
phosphorylation.
· ATP depletion: ouabain-sensitive Na+,K+-ATPase in plasma
membrane stops working, Na+ accumulates intracellularly, cell swells;
cell switches to anaerobic metabolism (glycolysis) and glycogen stores
are depleted, pH goes down.
· Oxygen and oxygen-derived free radicals/reactive oxygen species
· Loss of intracellular calcium homeostasis: activates phospholopases,
proteases, ATPases, endonucleases
· Defects in membrane permeability
· Irreversible mitochondrial damage: leakage of cytochrome c
triggering apoptotic cell death
• Reversible cell injury: cell swelling, detachment of ribosomes from
granular e.r. and dissociation of polysomes into monosomes. Fatty
change encountered in cells invloved in fat metabolism (hepatocyte,
myocardium). Histologically characterized by pallor, hydropic change,
vacuolar degeneration. EM: plasma membrane blebbing, blunting,
villous distortion, myelin figures, mitochondrial swelling, rarefaction,
nuclear disaggregation of granular and fibrillar elements.
Irreversible cell injury: mitochondria swell, lysosomes swell, damage
to plasma membrane and lysosomal membranes leads to enzyme
leakage; acidosis somewhat protective by inhibiting enzymatic
reactions.
Ischemia/Reperfusion Injury: new damage on reperfusion mediated
by oxygen free radicals and cytokine/adhesion molecules furthering
immune-mediated injury.
• Free radicals: def. chemical species with a single unpaired electron in
outer orbit.
Cause injury by: lipid peroxidation of membranes, oxidative
modification of proteins, lesions in DNA (single strand breaks).
Inactivated by: antioxidants (vit A, E, glutathione), binding to storage
and transfer proteins (transferrin, ferritin, lactoferrin, ceruloplasmin),
enzymes (catalase, SOD, glutathione peroxidase)

Chemical Injury:
mercuric chloride - binds sulfhydryl groups of proteins
cyanide - poisons mitochondrial cytochrome oxidase
CCl4 - conversions to free radical CCl3· causing lipid peroxidation
Acetaminophen - P450 catalyzed oxidation to toxic metabolite
CELL DEATH:
• Necrosis: spectrum of morphologic changes that follow cell death in living tissue
that result from progressive degradative action of enzymes on the lethally injured
cell. Morphologic appearance of necrosis is the result of enzyme digestion &
denaturation of proteins. Histologically: increased eosinophilia, karyolysis (nuclear
pallor), pyknosis (nuclear shrinkage), karyorrhexis (nuclear fragmentation).
Types:
1. Coagulative: e.g. myocardial infarct
2. Liquefactive: e.g. bacterial or fungal infections, CNS hypoxia
3. Gangrenous: e.g. limb ischemia (usually a combination of coagulative and
liquefactive necrosis), surgical term
4. Caseous: e.g. tuberculosis. characterized by granular debris w/obliteration of
tissue architecture (gross: white & cheesy)
5. Fat necrosis
Coagulative necrosis:
Liquefactive necrosis
Gangrenous necrosis
Casseous necrosis:
Fat necrosis:
•
Apoptosis:  programmed cell death.
Chracterized by cell shrinkage, chromatin condensation, formation of cytoplasmic blebs and
apoptotic bodies, phagocytosis by macrophages or adjacent cells. Does not elicit
inflammation (in contrast to necrosis).
Biochemical features:
1. Protein cleavage (caspases)
2. Protein cross-linking
3. DNA breakdown (endonuclease)
4. Phagocytic recognition (mediated by phosphatidylserine and thrombospondin expression)
Cytogenetic features:
1. ced genes
2. Fas-Fas ligand model: CD95 receptor on cell surface, TNF and TNFR-mediated apoptosis
3. Bcl-2: supresses apoptosis by direct action on mitochondria (preventing increased
permeability) & by binding other proteins (Apaf-1)
4. caspase (cysteine proteases that cleave after aspartic acid) mediated proteolytic cascade