الفهرس 
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Abstract
This work is aimed to biochemically investigate the pathogenic
mechanisms of tubulointerstitial changes due to induced injury in kidney
using mouse as an animal model.
This aim will be approved using different technical steps as:
1. Establishment of the animal model of renal tubulointerstitial injury
induced by unilateral ureteral obstruction (UUO).
2. Investigations Provided to Measure Renal Injury.
The results of the present work study different parameters where
this model combines several key advantages for studying development
and progression of kidney injury, including assessment of functional
consequences of kidney injury using biochemical evaluation in blood
such as blood urea nitrogen (BUN), serum creatinine measurements and
the ability to study pathophysiology during kidney injury through
morphometric evaluation of interstitial fibrosis within groups at different
UUO sacrifice time 3, 7 and 14 days to show the developments during
kidney injury in glomerular infiltration, interstitial inflammation,
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interstitial fibrosis, necrosis and dilated tubules by using hematoxylene
and Eiosn (H & E) stain, special stain as MT representing fibrosis and
PAS representing and Immunohistochemical stain for TNF and collagen .
The work with this model showed biochemically the pathogenic
mechanisms of tubulointerstitial changes due to induced injury in kidney
using mouse as an animal model. The study used a group of healthy
C57BL/6 mice which were randomly chosen. Their ages were ranged
from 8- 10 weeks and their weights were in the range of 20- 23 grams.
Thirty five mice were used in these experiments where different
groups were recruited to investigate the different disease severity; they
were divided according to the time of mice sacrificed after UUO
induction. These times were 3, 7 and 14 days after UUO operation. Sham
operation were performed had their ureters manipulated but not ligated.
When investigations were measured to monitor changes
biochemically we found changes in both creatinine and BUN along
different UUO groups. Creatinine shows a significant decrease comparing
control group within 3 and 7 days then rise in 14 days but less than
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control. While BUN shows a significant increase through 3, 7 and 14
days when compare with control groups.
After the histopathologically examination different markers have
been studied, monitoring the way to develop fibrosis and kidney injury.
By using hematoxylene and Eiosn (H & E) stain at different UUO
sacrifice time. After 3 days post operation, the kidney shows
hypercellularity of mesangial cells with marked lymphohistiocytic
exudate in interstitial tissue, dilation of renal glomeruli and interstitial
nephritis with fibroblastic proliferation. Kidney shows degenerative
change in renal tubules with histiocytic infiltrations in interstitial tissue,
severe congestion in interstitial blood capillaries with cystic dilatation of
renal tubules and loss of renal tubules.
After 7 days post operation, the kidney shows lymphohistiocytic
exudate infiltrate interstitial tissue with mild fibroblastic proliferation and
cystic dilation of renal tubules, hypercellularity of activated mesangial
cells with dissolution of renal glomeruli, proliferative glomerulonephritis
with formation of epithelial cresent and congestion of glomerular
capillaries, marked hemorrhage in interstitial tissue. Although kidney
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shows mononuclear cell infiltrates the interstitial tissue with fibroblastic
proliferation.
After 14 days post operation, there is massive lymphohistiocytic
infiltration interstitial tissue with replacement of the renal parenchyma,
extensive replacement of renal tissue with chronic inflammatory exudate
and fibroblastic proliferation in interstitial tissue with degeneration of the
renal parenchyma.
By using special stains at different UUO sacrifice time. MT
representing fibrosis developed by blue color as Special stain, which
shows different color intensities according to the fibrosis degree that
developed by blue color intensities in interstitial tissue which observed
from 3 days and increase gradually in 7 days till become thicker in 14
days as extensive fibrous tissue proliferation.
PAS stain shows glycogen precipitated in basement membrane
which lead to kidney dysfunction, it shows gradually increase in glycogen
precipitated in basement membrane intensities in dependent manner with
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different UUO sacrifice time leading to a complete structures and
function loss.
By using immunohistochemical stain at different UUO sacrifice
time groups for TNF that representing by brown color intensities that
increase gradually within at different time after UUO operation
confirming renal injury. Collagen also monitored by immunostaining is
representing renal damage. It also shows gradually increase in collagen
intensities in dependent manner with different UUO sacrifice time
leading to a complete structures and function loss
In conclusion this work show that C57BL/6 mice can be taken as
a good model for studying the mechanism of UUO changes while a
typical biochemical morphological and immunohistochemical changes
have been provided through different parameters measured in each item.
With this model we may trying a treatment concerning the different
changes we have been found during the study.