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العنوان
Effect of oral insulin -Chitosan Nanoparticles Versus Injected Insulin on Induced Diabetic Nephropathy in Male Albino Rats /
المؤلف
Mohammed, Wafaa Rabee.
هيئة الاعداد
باحث / وفاء ربيع محمد
مشرف / هــدى فؤاد ندا
مشرف / سوميه عبد العليم محمد
مشرف / منال شعبان حافظ
مشرف / عبير عبدالمحسن عبد الصمد
تاريخ النشر
2017.
عدد الصفحات
212 p. :
اللغة
الإنجليزية
الدرجة
الدكتوراه
التخصص
علم الأنسجة
تاريخ الإجازة
1/1/2017
مكان الإجازة
جامعة عين شمس - كلية الطب - علم الأنسجة
الفهرس
Only 14 pages are availabe for public view

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Abstract

as a loss of glycemic control. Insulin is the drug therapy for patients with insulin-dependent diabetes mel¬litus (or type 1 diabetes), and it is an important hormone for the regulation of blood glucose level. However, the problem encountered with subcutaneous insulin injections are pain, allergic reactions, hyperinsulinemia, and insulin lipodystrophy around the injection site.
The development of oral insulin loaded in nanoparticles is very essential to overcome the problem of insulin degradation in the stomach when administered orally.
Among the natural polymers used for oral insulin delivery is chitosan (CS) (a derivative of chitin, a natural structural polymer found in crustaceans and fungi).It is widely used owing to its ease of chemical modification and favorable biological properties.
The aim of the present study was to prepare and characterize oral insulin- chitosan nanoparticles and to compare the effect of oral insulin and injected insulin on the structure of the kidney in sterptozotocin-induced diabetic male albino rats.
Sixty-five adult male albino rats of average weight 200 grams were included in this study. The animals were divided into three groups:
group I (Control group): It included 25 rats. They were divided as follows:
Subgroup Ia: Five rats were the negative control group. Animals were left without intervention.
Subgroup Ib: It included ten rats. Each rat received single intraperitoneal injection of 0.5 ml streptoztocin vehicle. Five animals were sacrificed after four weeks. The others were sacrificed after six weeks.
Subgroup Ic: It included five rats. Each rat was subjected to single subcutaneous injection of 0.5ml saline daily for two weeks.
Subgroup Id: It included five rats.Each rat received 3ml oral chitosan-tripolyphosphate by gastric tube daily for two weeks.
group II(Diabetic group): It included 20 rats; each rat was injected once by streptozotocin (STZ) intraperitoneally in a dose of 60 mg/ kg B.W dissolved in 0.5ml of 0.1M citrate buffer. Animals were considered to be diabetic with blood glucose level >250mg/dl. Then, the animals were subdivided into two subgroups, ten animals each:
Subgroup IIa: Animals were sacrificed four weeks after induction of diabetes.
Subgroup IIb: Animals were sacrificed six weeks after induction of diabetes.
group III (treated group): It included 20 rats; in each rat induction of diabetes was done as group II and then they were left for four weeks from induction of diabetes after that the animals were further subdivided into two equal subgroups (ten rats each):
Subgroup IIIa: Animals were injected daily subcutaneously by insulin for another two weeks. The dose was 5 I.U/ rat /day.
Subgroup IIIb: Animals received oral insulin loaded on chitosan -tripolyphosphate nanoparticles dissolved in distilled water once daily by gastric tube for another two weeks. The dose was 100IU/kg/day.
Characterization of oral insulin:
Characterization of insulin loaded chitosan nanoparticles was done by using transmission electron microscopy(TEM), X-ray diffractometer (XRD), Nano Zeta sizer particle analyzer and Fourier transform infrared spectroscopic analysis(FTIR).
The experimental animals of each group were sacrificed after ether inhalation. The kidneys were excised, one half of each kidney was fixed in 10% formaldehyde saline and processed for light microscopic examination. Sections were stained with H&E, PAS and Mallory’s Trichrome stain. Small specimens from the other half were fixed in 3% gluteraldhyde and prepared for transmission electron microscopic examination. Morphometrical measurements and statistical analysis were also done.
In the current study, H & E stained sections of the renal cortex of subgroup IIa (4 weeks after induction of diabetes) revealed focal affection of glomeruli. Some of the glomeruli were enlarged with narrowing of their Bowman’s space. The cells of glomerular capillaries had pyknotic nuclei. Renal tubules showed variable degrees of affection. The cells lining some of the tubules showed vacuolated cytoplasm and pyknotic nuclei. Other tubules were disorganized and showed sloughing of their epithelium.
With prolonged untreated hyperglycemia, the renal cortex of subgroup IIb (6weeks after induction of diabetes) showed progressive damage. Most of the glomeruli were hypertrophied with obliteration of Bowman’s spaces. Destruction of the parietal layer of Bowman’s capsule of some renal corpuscles was also observed. Vacuolation in the cytoplasm of cells of glomerular capillaries and pyknosis of their nuclei were also detected. Proximal and distal convoluted tubules were severely affected compared to subgroup IIa. Some cells lining the tubules had deep acidophilic cytoplasm. Their lumina were wide and contained cellular debris, other tubules appeared hyalinized.
Injection of insulin in subgroup IIIa was less effective than oral insulin (IIIb) in improvement of renal damage. In subgroup IIIa some of the renal corpuscles showed hypertrophied glomeruli with narrow Bowman’s spaces and others were atrophied with widening of Bowman’s spaces. The cells lining some DCTs showed pyknotic nuclei. Cellular debris were still observed in the lumen of some tubules.
Oral insulin improved the renal structure in group IIIB, the kidney parenchyma retained nearly a histological structure comparable to the control group.
By PAS stain, in the diabetic subgroups the basement membrane of both glomerular capillaries and renal tubules showed increased PAS positive reaction. There was significant increase in the thickness of tubular basement membranes in comparison to the control group. Partial or complete loss of the brush border of PCTs was also noticed.
In addition in subgroup IIb, strong PAS positive exudates was noticed in the lumen of some tubules and in-between cortical and medullary tubules.
In subgroup IIIa, interrupted PAS reaction in the brush border of some PCTs was still observed. While in oral insulin (subgroup IIIb), PAS stained sections were relatively comparable to the control group.
By Mallory’s trichrome stain, the diabetic subgroups (IIa &IIb) showed a significant increase in mean area percentage of collagen fibers deposition in comparison to the control group. But after treatment by insulin, there was significant decrease (P<0.05) in mean area percentage of collagen fiber deposition in both subgroups IIIa& IIIb as compared with the diabetic subgroups.
The present study confirmed that a novel NP system composed of low MW CS and TPP was successfully prepared. Additionally, this work clearly demonstrated that insulin loaded NPs could effectively reduce the blood glucose level and improved the histological structure of the kidney in a diabetic rat model. So, oral insulin could be more effective than insulin injection in the treatment of diabetic nephropathy.