טכניון מכון טכנולוגי לישראל
הטכניון מכון טכנולוגי לישראל - בית הספר ללימודי מוסמכים  
M.Sc Thesis
M.Sc StudentToukan Hala
SubjectDetection and Localization of Endothelin Converting
Enzyme (ECE)-1 in the Human Diabetic Kidney
DepartmentDepartment of Medicine
Supervisor Clinical Professor Zaher Azzam
Full Thesis text - in Hebrew Full thesis text - Hebrew Version


Abstract

Introduction: Diabetes is a common chronic disease, accompanied with micro and macrovascular complications. The kidney is an important target organ injured after several years of uncontrolled diabetes.  The basis of renal injury is vascular damage and endothelial dysfunction. 

Aims: This study was designed to (a) examine the Endothelin Converting Enzyme (ECE)-1 protein expression in human diabetic kidney, and to detect its location. (b) examine the ETA and ETB receptors (ETAR and ETBR, respectively) protein expression in human diabetic kidney, and to detect their location.

Methods: Kidney samples were obtained from patients undergoing nephrectomy surgery. Samples were collected from various parts of the kidney: cortex, medulla and papilla, and used to produce protein. Additional samples were used for preparing sections for the immunohistochemical staining.

Western blot analysis was used to determine the expression level of the different proteins, and immunohistochemistry method was used for determining the location of the proteins in kidney cells.

Results: in the different regions of the kidney: cortex, medulla and papilla, an increase in the ECE-1 expression was obtained in the different study groups (diabetic and impaired fasting glucose- IFG), as compared to the control non diabetic group.

In the cortex, an increase in the expression of ETAR was obtained in the diabetic group as compared to IFG. A decrease in the protein expression was found in the IFG group as compared to control.  In the medulla and the papilla there was an increase in ETAR expression in the diabetic group as compared to control.

In the cortex and the papilla, a decrease in the levels of ETBR was obtained in the diabetic group as compared to control. In the medulla, a decrease in ETBR expression was found in the diabetic group as compared to control (based only on the western blot results).


Conclusions: Diabetes causes changes in the levels of the enzyme ECE-1, and endothelin receptors. Previous studies have been done on rats and other experimental animals. whereas, our study included patients. It is very important to learn about the effects of diabetes on the levels of the enzyme and the different receptors, because of the changes that happen under different clinical situations.