الفهرس 
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Abstract
The current study was undertaken to address the therapeutic effects of
mesenchymal stem cells (BM-MSCs and AD-MSCs) on AKI induced
experimentally by cisplatin and tracing the mechanisms by which they act.
Also, the study was extended to optimize the dosage of MSCs infusion and
to explore the advantages of MSCs therapy over the conventional therapy
(Losartan) in the treatment of AKI.
For MSCs isolation and propagation; BM-MSCs were isolated from
bone marrow of both femur and tibia of male Wistar rats. While, ADMSCs
were obtained from abdominal fats of male Wistar rats. Then, the
isolated cells were grown and propagated in culture. To confirm that the
cells in the culture were MSCs, they were characterized morphologically
by inverted microscope examination, and by detection of their specific cell
surface markers (CD29, CD166, CD45, and CD34) gene expressions
which was done by conventional PCR technique.
The in vivo study was conducted on 90 adult male Wistar rats which
assigned into 9 groups (10 rats/group). group 1: Control. group 2: AKI, in
which rats received a single intraperitoneal injection of cisplatin (6 mg/
kg). group 3: Losartan, in which AKI-induced rats received losartan
dissolved in saline orally in a dose of 10 mg/ kg/ day for two months.
Groups (4 & 5& 6): AKI groups treated with BM-MSCs (1x106cells/rat,
2x106cells/rat and 4x106cells/rat, respectively). Groups (7 & 8 & 9): AKI
groups treated with AD-MSCs (1x106cells/rat, 2x106cells/rat and
4x106cells/rat, respectively).
To evaluate the therapeutic efficacy of MSCs either derived from
bone marrow or adipose tissue against AKI, the ability of the injected
MSCs to migrate to the injured kidneys was confirmed by labeling of
MSCs with PKH-26 dye. Serum creatinine, urea, MDA levels as well as SOD and CAT activities were assayed by colorimetric technique. Urinary
(KIM-1 and IL-18), serum (Cys C, TNF-α, MCP-1, MIP-2 and NQO-1)
were detected by ELISA technique. Whereas, kidney p38, Bcl-2 and
VEGF expressions were determined by conventional PCR. Additionally,
histopathological investigation of kidney tissues was carried out.
The results of the study showed that MSCs were present in the kidney
tissues of AKI rats treated with BM-MSCs or AD-MSCs indicating that
the delivered undifferentiated MSCs were able to home at the injured
tissues. It is clearly observed from the biochemical analyses that cisplatin
injection caused significant increase in creatinine, urea, cys C, KIM-1,
TNF-α, MCP-1, MIP-2, IL-18 and MDA levels associated with significant
decrease in SOD, CAT activities as well as NQO-1 level. Also, cisplatin
produced up- regulation of p38 gene expression and down- regulation of
Bcl-2 and VEGF mRNA. Moreover, the kidney tissue sections of AKI
bearing rats showed coagulative necrosis in diffuse manner all over the
tubular lining epithelium with large number of apoptotic cells, associated
with swelling in the endothelial cells lining the tufts of the glomeruli as
well as marked inflammatory infiltrate involving most of the renal tubules.
On the opposite side, all treatments (Losartan, BM-MSCs and ADMSCs)
produced significant decrease in the levels of creatinine, urea, cys
C, KIM-1, TNF-α, MCP-1, MIP-2, IL-18 and MDA associated with
significant increase in SOD, CAT activities and NQO-1 level. Also, there
is significant down- regulation in p38 gene expression and up- regulation
in Bcl-2 and VEGF mRNA. Furthermore, the histopathological findings
were as follows: In losartan treated group, there was large number of
mononuclear cells infiltrates many tubules, esinophilic casts in some
tubules and numerous apoptotic bodies. There was moderate inflammatory
infiltrates and many apoptotic cells observed in group treated by BM MSCs (1 ×106 cells rat). In BM-MSCs (2×106 cells rat)-treated group,
congested blood vessels, interstitial hemorrhage as well as few
inflammatory cells and some apoptotic cells were noticed. BM-MSCs
(4×106 cells rat)-treated group showed well-formed tubules and very few
apoptotic cells. In AD-MSCs (1×106 cells rat)-treated group, congested
blood vessels, hyaline casts, minimal inflammatory infiltrates and some
apoptotic cells were detected. AD-MSCs (2×106 cells rat)-treated group
showed localized collection of inflammatory cells and few apoptotic
bodies. While, AD-MSCs (4×106 cells rat)-treated group denoted wellformed
glomeruli and tubules with no histopathological alterations.