الفهرس 
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Abstract
Some recent studies have found a way to treat kidney damages caused by certain toxic materials.
BM-MSCs have a great ability to differentiate into different functional cells in addition to their ease isolation, homing ability as well as anti-apoptotic potentials made them the optimal treatment for many incurable diseases.
Cisplatin is the most potent anticancer drugs, however, there is a limitation in its clinical use in the last few years due to organs cytotoxicity including nephrotoxicity owing to its accumulation in the renal cells leading to renal damage.
Hesperidin is a natural antioxidant flavonoid that exerts antioxidant, anti-inflammatory as well as anti-apoptotic properties.
Therefore, the current study was designed to evaluate the ameliorating effect of BM-MSCs and hesperidin either singly or together on cisplatin-induced nephrotoxicity in rats.
Fifty male Wistar rats were used in the present study and allocated into five equal groups as follows:
1-group I (control group): animals of this group received (CMC) 1%, once daily via oral gavage for 4 weeks as well as a single dose of intraperitoneal injection of 1ml normal saline (0.9% Nacl).
2-group II (Cisplatin administered group): this group was given a single dose of intraperitoneal injection of cisplatin 7.5mg/kg body weight.
3-group III (Hesperidin treated group): the rats of this group were administered hesperidin orally by a dose of 40 mg/kg dissolved in CMC 1% once daily for four weeks. On the 5th day of hesperidin administration, cisplatin (7.5 mg/kg) was taken as a single dose an hour before Hesperidin dose.
4-group IV (BM-MSCs- treated group): this group was received a single dose of intraperitoneal injection of cisplatin 7.5 mg/kg body. On the next day, rats were given a single intravenous dose of mesenchymal stem cells at dose of 1×106 cells/rat in the tail vein once per week for four weeks.
5-group V (Hesperidin and BM-MSCs-treated group against cisplatin-induced nephrotoxicity): Cisplatin administrated rats were treated with both hesperidin orally at a dose of 40 mg/kg daily for 4 weeks and intravenous injection of BM-MSCs at a dose of 1×106 cells/rat once per week for four weeks.
The BM-MSCs used in the current study were characterized morphologically by using inverted microscope which showed a fusiform shape with short cytoplasmic processes, also by RT-qPCR to confirm that the used cells were already stem cells, that revealed by the strong gene expression of BM-MSCs surface markers (CD73 and CD105) and the weak gene expression of hematopoietic surface markers (CD34 and CD45).
At the end of the experiment (after four weeks), rats were killed and blood samples were obtained for detection of the biochemical renal parameters (serum creatinine, urea, sodium and potassium levels), the inflammatory cytokines (NF-κB in serum) and (IL10 and TNF-α in renal Tissue Homogenate). For further histopathological and immunohistochemical studies, the kidneys were taken and stained by several stains as H&E, PAS, bromophenol blue as well as P53-immunohistochemical technique for apoptosis detection.
The light microscopic examination of the kidney tissue of the cisplatin-administrated rats revealed degenerative changes in renal corpuscles and tubules, the renal corpuscles appeared with atrophied glomeruli, wide Bowman’s space and showed partial loss of the parietal layer of Bowman’s capsule. Also, most tubular epithelial cells showed pyknotic nuclei, cytoplasmic vacuolation and hydropic degeneration in addition to interstitial lymphocytic infiltration as well as severe congestion of the glomerular tuft and peritubular capillaries. Some renal tubules showed cystic dilatation and distended with granular and hyaline materials. On the level of histochemical reactions, a marked reduction of mucopolysaccharides and total protein content in tubular cytoplasm were observed, in addition, the kidney sections exhibited strong immunoreactivity to P53 antibody.
The kidney sections of hesperidin treated rats showed that most renal corpuscles had intact glomeruli and bowman capsules, the epithelial cells of the proximal and distal convoluted tubules exhibited few cytoplasmic vacuolation and less hydropic degeneration with normal vesicular nuclei. Moderate improvement in mucopolysaccharides and total protein content in the renal section were found. However, areas of interstitial lymphocytic infiltration and vascular congestion were detected. The renal sections exhibited a moderate immunoreactivity to P53antibodies in the tubular cells.
In the BM-MSCs treated group, a marked restoration of a normal intact renal corpuscle and improvement of cytoplasmic degeneration and vacuolation, nuclei became vesicular with marked decrease of inflammatory cells were observed. A marked improvement of cellular mucopolysaccharides and total protein content in renal tissue were noticed. However, some congestion in glomerular tuft and other blood vessels were observed, the renal sections exhibited a weak immunoreactivity to P53 antibody.
In BM-MSCs and hesperidin treated group, microscopic examination showed complete regeneration of the renal tissue that looked similar to that of the control group. The renal corpuscles appeared with normal glomeruli and renal tubules with a complete absence of lymphocytic cells’ infiltration. Moreover, pronounced improvement in total protein content and mucopolysaccharides in the renal tissue resemble that of the normal kidney as well as barely detectable reaction with P53 antibody.
Concerning the biochemical reactions, the rats administrated cisplatin showed a marked disturbances in biochemical parameters demonstrated by significant increase in urea, creatinine and potassium and lowered sodium serum levels when compared with control rats, while the treatment of cisplatin-administered rats with BM-MSCs and/or hesperidin showed a significant improvement in serum renal parameters when compared with cisplatin administrated group.
As for serum inflammatory cytokines levels, this study also targeted the effect of BM-MSCs and/or hesperidin on cisplatin-induced nephrotoxicity in rats, cisplatin administration revealed a significant increase of inflammatory cytokines TNF-α in the renal tissue homogenate and NF-κB in serum and decrease of anti-inflammatory cytokine (IL-10) in the renal tissue homogenate when compared with control group. In contrast, the treatment of cisplatin administrated rats either with BM-MSCs and hesperidin singly or in combination is significantly decreased (TNF-α and NF-κB) and increased IL-10 levels compared to cisplatin administrated group.