الفهرس 
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Abstract
The aim of this study was to evaluate the possible protective role of Moringa oleifera versus omega-3 fatty acids on the effect of high fat diet (HFD) on the renal cortex of adult male rats.
The experiment was conducted on sixty adult male albino Wistar rats, aged 8 weeks and weighing 180-200 gm. Rats were randomly divided into four groups and were fed their specified diets for four weeks as follow:
• group I (control group): It included thirty animals which were subdivided into three equal subgroups (ten rats each):
o Subgroup Ia: rats were fed the standard diet.
o Subgroup Ib: rats were fed the standard diet in addition to Moringa oleifera (MO) aqueous extract at a dose of 1000 mg/kg/day through intragastric tube.
o Subgroup Іc: rats were fed the standard diet in addition to Omega-3 fatty acids (omega-3 FA) at a dose of 300 mg/kg/day through intragastric tube.
• group II (HFD): It included ten rats that were fed HFD.
• group III (HFD and MO): It included ten rats that were fed HFD in addition to MO aqueous extract at a dose of 1000 mg/kg/day through intragastric tube.
• group ІV (HFD and Omega-3 FA): It included ten rats that were fed HFD diet in addition to Omega-3 FA at a dose of 300 mg/kg/day through intragastric tube.
At the end of the experiment, all animals were weighed. Blood samples were taken to detect the level of fasting blood glucose, triglycerides (TG), cholesterol (CH), LDL, HDL, urea and creatinine. Then all animals were sacrificed and specimens from the kidneys were prepared for light and transmission electron microscopic examination. Histological techniques included H&E, Masson trichrome stain, PAS and Immunohistochemical staining for α-SMA.
The mean body weight of the animals of group II (HFD) revealed a significant increase as compared to group I. Meanwhile, the mean body weight of animals of groups III and IV was significantly decreased as compared to group II.
Concerning the biochemical results of the rats of group II, there was a significant increase in the mean level of fasting blood glucose, TG, CH, LDL, urea and creatinine. However, the mean level of HDL revealed a significant decrease as compared to group I. The biochemical results of the rats of groups III and IV revealed a significant decrease in the mean level of fasting blood glucose, TG, CH, LDL, urea and creatinine. However, the mean level of HDL revealed a significant increase as compared to group II. group III showed a non-significant difference as compared to group IV.
Light microscopic examination of H and E stained sections of the kidneys of group II (HFD) rats revealed focal structural changes in the renal cortex. Some renal corpuscles showed hypercellularity of glomeruli. Other corpuscles showed detached folded parietal layer of the Bowman’s capsule and focal adhesions with the capillary tuft and reduced Bowman’s space.
Some PCTs were disorganized having dilated lumina with apparent decrease in the height of some lining cells. Many of their lining cells showed deep homogenous acidophilic cytoplasm, while others showed vacuolated cytoplasm. Some DCTs appeared dilated with focal loss of the lining epithelial cells. Many nuclei of the tubular cells appeared pyknotic, while some appeared binucleated. The lumina of some renal tubules exhibited homogenous acidophilic material and exfoliated cells. The interstitium showed mononuclear cellular infiltration.
Masson’s trichrome stained sections of the rats of group II revealed a significant increase in the collagen fibers around the renal corpuscles and in the interstitium surrounding the renal tubules as compared to the control group.
PAS-stained sections of the rats of group II showed focal loss of the apical brush borders in the cells of some PCTs. The basement membrane of some renal tubules appeared disrupted. The basement membranes surrounding the glomerular capillaries and the renal corpuscles were apparently thickened.
Immunohistochemical staining of the section of the rats of group II showed a significant increase in the reaction for α-SMA as compared to the control group. Strong postive reaction was detected in the interstitium, while moderate positive reaction was detected in some glomeruli and the cytoplasm of some cells of the renal tubules.
By TEM, the kidney sections of the rats of group II showed apparent increase in the number of intraglomerular mesangial cells with apparent increase in the surrounding non-homogeneous mesangial matrix as compared to the control group. Podocytes were noticed with relatively fewer indistinct primary and secondary processes as compared to the control group. Fusion of the secondary processes of the podocytes was frequently discovered with reduction in the sub-podocytic space. The GBM showed apparent focal increased thickness as compared to the control group.
Some lining cells of the PCTs revealed shrunken indented nuclei, electron lucent vacuoles and multiple electron dense bodies within their cytoplasm. The cells also showed apparent increase in the number of mitochondria which appeared disorganized and smaller in size as compared to the control group. Some PCTs showed irregular electron dense basement membrane. Some DCT cells rested on irregular basement membrane. The cells had shrunken indented nuclei and irregularly arranged mitochondria with indistinct basal cell membrane infoldings.
Examined H and E stained sections of the kidneys of group III (HFD and MO) rats revealed more or less normal appearance of most of renal cortical tissue as compared to the control group. However, focal changes were still detected in the form of vacuolated cytoplasm of few tubular cells.
Masson’s trichrome stained sections of the rats of group III revealed a significant decrease in collagen content as compared to the HFD group and a non-significant difference as compared to the control group.
PAS-stained sections of the renal cortex of the rats of group III showed PAS positive apical brush borders of most of the cells of the PCTs. The basement membrane surrounding the glomerular capillaries, the renal corpuscles and the renal tubules showed a well-defined PAS positive reaction. However, the basement membrane around some tubules revealed a faint positive reaction.
Immunohistochemical staining for α-SMA of the sections of the renal cortex of the rats of group III revealed a significant decrease as compared to group II (HFD). However, there was a significant increased as compared to the control group. Scattered faint positive reaction was detected in the glomeruli and the interstitium, while the renal tubules showed negative reaction.
Transmission electron microscopic examination of the kidney sections of the rats of group III revealed a comparable ultrastructural profile as that of the control group. Intraglomerular mesangial cells were detected in between glomerular capillaries. They were less frequently observed and surrounded by apparently decreased mesangial matrix as compared to the HFD group. Podocytes were noticed with relatively more numerous primary and secondary processes extending towards the GBM as compared to the HFD group. Distinct filtration slits appeared between the secondary processes of the podocytes and were closed by diaphragms. Preservation of the sub-podocytic space was also noticed. The GBM revealed uniform thickness.
The PCTs and DCTs showed more or less comparable histological ultrastructure as that of the control group, however the PCTs cells showed apparent decrease in the number of mitochondria as compared to the HFD group.
Light microscopic examination of H and E stained sections of the kidneys of group IV (HFD and Omega-3 fatty acids) rats showed more or less normal structure of most of the renal cortical tissue as compared to the control group. However, in some sections, focally affected areas with structural alterations were detected. These areas showed vacuolated cytoplasm of some renal tubules. Other tubules showed dilated lumina and desquamated cells.
Masson’s trichrome stained renal sections of the rats of group IV revealed a significant decrease in the collagen fibers content as compared to the HFD group. However, there was a significant increase in the collagen fibers content as compared to the groups I and III.
PAS-stained sections of the renal cortex of the rats of group IV showed PAS positive apical brush borders of most of the cells of the PCTs. The basement membrane surrounding the glomerular capillaries, the renal corpuscles and most of the renal tubules showed a well-defined PAS positive reaction. However, in some sections, the PCTs showed focal loss of the apical brush borders. Ill-defined basement membrane of some tubules was also detected.
Immunohistochemical staining for α-SMA of the sections of the renal cortex of the rats of group IV showed a significant decrease as compared to the HFD group. However, there was a significant increase as compared to the groups I and III. Weak positive reaction was detected in some glomeruli, renal tubules and in the interstitium.
By TEM, the kidney sections of the rats of group IV revealed an improved ultrastructural profile as compared to the HFD group. The intraglomerular mesangial cells were more frequently observed in between glomerular capillaries as in the HFD group when compared to group I and group III. Podocytes were noticed with relatively numerous primary and secondary processes extending towards the GBM as compared to the HFD group. Filtration slits appeared between the secondary processes of the podocytes and were closed by diaphragms. Reduced sub-podocytic space was also noticed. The GBM revealed scattered focal increase in thickness as in the HFD group when compared to group I and group III.
The PCTs and DCTs revealed more or less comparable histological ultrastructure as that of the control group, however the PCTs cells showed apparently decreased number of mitochondria as compared to the HFD group.