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Journal of the Korean Society of Pediatric Nephrology 2004;8(2): 138-148.
당과 후기당화합물에 의한 사구체 상피세포 ZO-1 발현의 변화
이진석, 이해수, 윤옥자, 하태선
1충북대학교 의과대학 소아과과학교실
2충북대학교 의과대학 의학연구소
3충북대학교 의과대학 의학연구소
4충북대학교 의과대학 소아과과학교실
Effects of High Glucose and Advanced Glycosylation Endproducts (AGE) on ZO-1 Expression in cultured Glomerular Epithelial Cells (GEpC)
Jin-Seok Lee, Hae- Soo Lee, Ok-Ja Yoon, Tae-Sun Ha
1Department of Pediatrics, Chungbuk National University Hospital
2Medical Research Institute, Chungbuk National University Hospital
3Medical Research Institute, Chungbuk National University Hospital
4Department of Pediatrics, Chungbuk National University Hospital
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ABSTRACT
Purpose: Regardless of the underlying diseases, the proteinuric condition demonstrates ultrastructural changes in podocytes with retraction and effacement of the highly specialized interdigitating foot processes. We examined the molecular basis for this alteration of the podocyte phenotypes, including quantitative and distributional changes of ZO-1 protein as a candidate contributing to the pathogenic changes in the barrier to protein filtration.
Methods: To investigate whether high glucose and advanced glycosylation endproduct(AGE) induce podocyte cytoskeletal changes, we cultured rat GEpC under 1) normal glucose(5 mM=control) or 2) high glucose(30 mM) or 3) AGE-added or 4) high glucose plus AGE-added conditions. The distribution of ZO-1 was observed by confocal microscope and the change of ZO-1 expression was measured by Western blotting and RT-PCR.
Results : By confocal microscopy, we observed that ZO-1 moves from peripheral cytoplasm to inner actin filaments complexes in both AGE-added and high glucose condition. In Western blotting, high glucose or AGE-added condition decreased the ZO-1 protein expression by 11.1%(P>0.05) and 2.3%(P>0.05), respectively compared to the normal glucose condition. High glucose plus AGE-added condition further decreased ZO-1 protein expression to statistically significant level(12%, P<0.05). No significant change was seen in the osmotic control. In RT-PCR, high glucose plus AGE-added condition significantly decreased the expression of ZO-1 mRNA by 12% compared to normal glucose condition.
Conclusion : We suggest that both high glucose and AGE-added condition induce the cytoplasmic translocation and suppresses the production of ZO-1 at transcriptional level and these changes may explain the functional changes of podocytes in diabetic conditions.
Key words: Diabetic nephropathy | Advanced glycosylation endproducts | ZO-1 | Glomerular epithelial cells | Podocvte
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