NECROSIS & APOPTOSIS

DR HUZLINDA HUSSIN
DEPT OF PATHOLOGY
SEM 1 2011/12

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Cell die by one of 2 mechanisms:
1. Necrosis
2. Apoptosis

Apoptosis & necrosis have different
characteristics.

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Necrosis

The more common type of cell death after exogenous stimuli eg:
ischemia, hypoxia, chemical injury

Refers to a spectrum of morphologic changes that follow cell death
in living tissue, largely resulting from the degradative action of
enzymes on the lethally injured cells.

its most common manifestation is coagulative necrosis.

The morphologic appearance is the result of 2 concurrent
processes:
1. Enzymatic digestion of cell
 Autolysis (the catalytic enzymes derived from the lysosomes of the dead cells
themselves)
 Heterolysis (the catalytic enzymes derived from the lysosomes of the
immigrant leukocytes)
2. Denaturation of protein

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Morphology of the necrotic cells
 Increased eosinophilia (partly d/t loss of normal
basophilia imparted by the DNA & partly d/t increased
binding of eosin to the denatured intracytoplasmic
proteins).
 Glassy homogenous appearance (mainly d/t loss of
glycogen particles)
 Vacuolated cytoplasm- moth-eaten (d/t enzymatic
digestion of the cytoplasmic organelles)
 Calcification of dead cells.
 Nuclear changes (early)- appear in the form of 1 of
3 patterns:
1. Karyolysis- fade off of the basophilia of the DNA.
2. Pyknosis- nuclear shrinkage & increased basophilia
3. karyorrhexis.- fragmentation of the pyknotic or partially
pyknotic nucleus.

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Types of necrosis:
1. Coagulative necrosis
 When protein denaturation is the primary pattern
 Firm texture of the affected tissue
 Basic outline of the cell is preserved at least for some days
 Finally- removal of the cellular debris by fragmentation and phagocytosis
by scavenger white cells and proteolytic lysosomal enzymes by immigrant
white cells.
 Characteristic of hypoxic death of all tissues (eg: myocardial & kidney
infarct) except the brain.
2. Liquefactive necrosis
 When enzymatic digestion is dominant
 Characteristic of focal bacterial or occasionally fungal infections (stimulate
inflammatory cells)
 Also occur in the hypoxic death of cells within CNS.
 End result-liquid viscous mass (liquefaction completely digests the dead
cells) or pus (if the process has been initiated by acute inflammation)
 Gangrenous necrosis-usually applied to a hypoxic limb that undergone
coagulative necrosis & superimposed by bacterial infection. The
coagulative necrosis is modified by the liquefactive action of the bact & the
attracted leukocytes (so-called wet gangrene)

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 Myocardial infarction is a type of
coagulative necrosis. The general shape
and structure of the organ are
maintained. In this picture, on the top half
of the left ventricular wall, you can see
an area of hemorrhagic necrosis.

AMI with coagulative necrosis

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 Pulmonary abscess is a localized
area of liquefactive necrosis of the
lung.

breakdown of tissue has elicted the collection of

macrophages, who've come in to clean up the
debris left by lysis.
liquefaction in the brain 9
3. Caseous necrosis
 A distinctive form of coagulative necrosis
 Encountered most often in foci of tuberculous infection
 Macros- white &cheesy
 Micros-the necrotic focus composed of fragmented,
coagulated cells & amorphous granular debris, enclosed
within a granulomatous reaction (tissue architecture is
completely obliterated)
4. Fat necrosis
 Does not denote a specific pattern of necrosis
 It is rather to describe focal areas of fat destruction, typically
occur as a result of release of activated pancreatic lipases into
the substance of pancreas & peritoneal cavity (in acute
pancreatitis)
 Macros-chalky white areas (fat saponification)
 Micros- foci of shadowy outline of necrotic fat cells with
basophilic calcium deposits, surrounded by an inflamm
reaction

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 Caseous necrosis (top) and palisading histiocytes
A tubercular granuloma with central, caseous below it.
necrosis 11
 enzymatic fat necrosis, gross; it is the chalky granular
stuff.

Pancreatic fat necrosis characterized by

enzymatic fat necrosis surrounded by a zone
containing nuclear dust and an inflammatory cell
infiltrate.

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Apoptosis

A form of cell death designed to eliminate

unwanted, aged or potentially harmful cells
thru activation of a coordinated, internally
programmed series of events affected by a
dedicated set of gene products

However, it also occurs under pathologic
condition in which s/times accompanied by
necrosis

Overlaps & shares certain common mech
with necrosis
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It occurs in following general settings
(physiologic, adaptive & pathologic events):
Apoptosis in physiologic situations:
1. During embryogenesis, including implantation,
organogenesis, developmental involution &
metamorphosis.

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 APOPTOSIS: important in embryogenesis

Morphogenesis (eliminates excess cells):

Selection (eliminates non-functional cells):

2. Involution of hormone-dependent tissues upon
hormone withdrawal.
eg: endometrial cell breakdown during menstrual
cycle, ovarian follicular atresia in menapause,
regression of lactating breast after weaning.
3. Cell loss in proliferating cell proliferations.
Eg: B lymphocytes in germinal centers and epith
cells in intest crypts- to maintain a constant
number (homeostasis)
4. Elimination of potentially harmful self reactive
lymphocytes, either b4 or after they have
completed their maturation- to prevent
reactions against one’s own tissues.
5. Death of host cells that have served their useful
purpose. Eg neutrophils after acute inflamm. 16
 APOPTOSIS: important in embryogenesis

Immunity (eliminates dangerous cells):

Self antigen
recognizing cell

Organ size (eliminates excess cells):

 APOPTOSIS: important in adults
Tissue remodeling (eliminates cells no longer needed):

Apoptosis
Virgin mammary gland Late pregnancy, lactation Involution
(non-pregnant, non-lactating)

- Testosterone
Apoptosis

Prostate gland

Apoptosis in pathologic conditions:
1. DNA damage- due to radiation, hypoxia etc
2. Accumulation of misfolded proteins (due to
mutations in the genes encoding these
proteins or becoz of extrinsic factors)
3. Cell death in certain infections- eg: virus HIV,
Hep B,C.
4. Pathologic atrophy in parenchymal organs
after duct obstruction eg; pancreas, parotid
gld. 19

Morphology
 Cell shrinkage- small size of round or oval shaped,
single or in small clusters with dense eosinophilic
cytoplasm
 Chromatin condensation & at periphery (the most
characteristic feature), may break up (2 or more
fragments)
 Cytoplasmic blebs & apoptotic bodies (with or without
nuclear fragment)

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 Phagocytosis of apoptotic cells or bodies by
adjacent healthy cells (parenchymal cells or
macrophages) & degraded within lysosomes.

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Biochemical features:
1. Protein cleavage/ hydrolysis
 A specific feature of apoptosis
 Involves the activation of several members of newly discovered
family of cystein proteases named caspases
 Caspases cleavage of the nuclear scaffold & cytoskeletal
proteins underlies the distinctive nuclear & cytoplasmic
structural alterations
2. Protein cross-linking
 By transglutaminase activation that converts cytoplasmic
proteins into covalently linked shrunken shells that may break
into apoptotic bodies.

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3. DNA breakdown
4. Phagocytic recognition (of apoptotic cells)
 Apoptotic cells express phosphatidylserine in the
outer layers of their plasma membrane, the
phospholipid having flipped out from the inner
layers
 Some have thrombospondin (an adhesive
glycoprotein) on the surface.
 Cells are disposed with minimal compromise to the
surrounding tissue (without the release of
proinflammatory cellular response)

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Mechanisms of apoptosis- consists of 4 separable but
overlapping components:
1. Signaling pathways- initiate apoptosis
a. transmembrane/external signals
o Negative or positive determinants
o Eg:
a. stimuli such as growth factors, certain hormones & cytokines
generate signal that suppress preexisting death programs but
the absence leads to failure of death suppression and thus
triggers apoptosis.

b. Receptor-ligand interactions at the plasma membrane that

generate signals to activate death programs- the most important
is tumour necrosis factor receptor (TNFR) superfamily.

b. Intracellular/internal signals
o Eg: binding of glucocorticoids to nuclear receptors;
physicochemical agents eg heat, radiation, hypoxia; hypoxia
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2. Control and integration stage
 In which intracellular positive & negative
regulatory molecules inhibit, stimulate or
forestall apoptosis & thus determine the
outcome.
 Performed by specific proteins that connect
death signals to the execution phase
 These proteins may result in either life or death
of cells- can affect the outcomes of dz
 2 broad schemes for this stage:
i. Direct transmission of signals by specific adapter
proteins to the execution mechanismc (as described
for the Fas-Fas ligand model) and target cell killing
by cytotoxic T lymphocytes.
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 APOPTOSIS: control
Receptor pathway (physiological):
FAS ligand TNF

Death receptors:
(FAS, TNF-R, etc)
Death
domains

Adaptor proteins

Pro-caspase 8 (inactive) Caspase 8 (active)

Pro-execution caspase (inactive)

Execution caspase (active)

MITOCHONDRIA Death
ii. Involves members of Bcl-2 family of proteins-
play major roles in apoptotic regulation largely
by regulating mitochondrial function.
 Death agonists can generate signal that affect
mitochondria in 2 ways:
i. Apoptotic signals results in mitochondrial permeability
transitions. Formation of pores within the inner
mitochondrial membrane results in reduction of
membrane potential & mitochondrial swelling.
ii. The signals also cause increased permeability of outer
mitochondrial membrane, releasing an apoptotic trigger
i.e cytochrome c.

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 APOPTOSIS: control
Intrinsic pathway (damage):

Mitochondria

BAX Cytochrome c release BCL-2

BAK BCL-XL
BOK BCL-W
BCL-Xs Pro-caspase 9 cleavage MCL1
BAD BFL1
BID DIVA
B IK NR-13
BIM Pro-execution caspase (3) cleavage Several
NIP3 viral
BNIP3 proteins

Caspase (3) cleavage of cellular proteins,

nuclease activation,
etc.

Death
 APOPTOSIS: control

Physiological Intrinsic
receptor pathway damage pathway

MITOCHONDRIAL SIGNALS

Caspase cleavage cascade

Orderly cleavage of proteins and DNA

CROSSLINKING OF CELL CORPSES; ENGULFMENT

(no inflammation)
3. The execution phase

Consisting of the actual death program &
accomplished largely by the caspase family of
proteases.

4. Removal of dead cells by phagocytosis

Apoptotic cells & their fragments hv marker
molecules on their surface- facilitates early
recognition by adjacent cells or phagocyes for
phagocytic uptake & disposal.

Efficient process-dad cell disappear without
leaving a trace & no inflammation.
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 Dysregulated apoptosis (too
litle or too much)
1. d/o associated with inhibited apoptosis &
increased cell survival.
a) Cancers- esp with p53 mutations or
hormone dependent tumours eg: breast,
prostate, ovary
b) Autoimmune d/o- could arise if autoreactive
lymphocytes are not removed after an
immune response.

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2. d/o ass with increased apoptosis & excessive
cell death:
a) Neurodegenerative dz eg: spinal muscular
atrophies- loss of specific sets of neurons
b) Ischemic injury eg: MI & stroke
c) Virus-induced lymphocyte depletion eg: AIDS

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Features of apoptosis vs.
necrosis

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