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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, Chapter6 Summary and Conclusions 85 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 Chapter6 Summary and Conclusions 86 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 Chapter6 Summary and Conclusions 87 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 Chapter6 Summary and Conclusions 88 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. |