Role of Antibody Anti-AGE in the Expression of Nephrin and RAGE on Primary Glomerulus Cell Exposed with AGE

Rudy Salam, Diana Lyrawati, Nur Samsu


Upregulation of nephrin expression occurs at the early stage of nephropathy and decrease in the period 6 months on hyperglycaemic condition. Nephrin is associated with the initial stage of the loss of the permeability barrier in diabetic nephropathy. Interaction AGE-RAGE increases angiotensin II on Renin Angiotensin-Aldosterone System (RAAS) and activation of protein kinase c (PKC) which induce alterations in nephrin mRNA expression. Alterations of nephrin expression induces transformation of slit membrane structure and the permeability changes at the glomerular filtration barrier. Anti-AGE vaccination once may cause the changes of nephrin and RAGE expression and can prevent progression of diabetic nephropathy. This study used primary glomerulus cell culture obtained from renal of Wistar mice aged 3 months, weighting 200-300 grams that consist of negative control group that exposed to BSA 100 µg/ml, positive control group that exposed to AGE-BSA 100 µg/ml, treatment group 1 that exposed to polyclonal anti-AGE 5 µg/ml and AGE-BSA 100 µg/ml and treatment group 2 that exposed to monoclonal antibody anti-CML 5 µg/ml and AGE-BSA 100 µg/ml. Paired t-test with a 0.05 level of confidence results showed that there were significant differences in level of RAGE expression among experimental groups with control groups. Administration of polyclonal antibody decreased RAGE expression among negative control (p=0.188). but not in positive control (p=0.000). In contrast to monoclonal anti-AGE antibody, RAGE expression did not differ significantly compared to negative control but significant than positive control. Administration of monoclonal anti-AGE antibody inhibited increasing of nephrin expression compared to negative and positive control (p=0.73; 0.125). In conclusion, this study suggested that administration of polyclonal and monoclonal anti-AGE antibody could inhibit increasing of RAGE and nephrin expression in glomerulus primary culture that exposed to AGE which is expected to prevent the progression of diabetic nephropaty.


anti-AGE antibody; AGE; RAGE; nephrin;primary glomerulus cell culture

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Aaltonen P, Luimula P, Aström E et al. (2001) Changes in the expression of nephrin gene and protein in experimental diabetic nephropathy. Laboratory Investigation 81 (9): 1185-1190.

Maezawa Y, Takemoto M, Yokote K (2015) Cell biology of diabetic nephropathy: Roles of endothelial cells, tubulointerstitial cells and podocytes. Journal of Diabetes Investigation 6 (1): 3-15. doi: 10.1111/jdi.12255.

Jia J, Ding G, Zhu J et al. (2008) Angiotensin II infusion induces nephrin expression changes and podocyte apoptosis. American Journal of Nephrology 28 (3): 500-507. doi: 10.1159/000113538.

Menne J, Meier M, Park JK et al. (2006) Nephrin loss in experimental diabetic nephropathy is prevented by deletion of protein kinase C alpha signaling in-vivo. Kidney International 70 (8): 1456-1462. doi: 10.1038/

Li X, Chuang PY, D'Agati VD et al. (2015) Nephrin preserves podocyte viability and glomerular structure and function in adult kidneys. Journal of the American Society of Nephrology 26 (10): 2361-2377. doi: 10.1681/ASN.2014040405.

Wang SX, Menè P, Holthofer H (2001) Nephrin mRNA regulation by protein kinase C. Journal of Nephrology 14 (2): 98-103.

Goldin A1, Beckman JA, Schmidt AM, Creager MA (2006) Advanced glycation end products: Sparking the development of diabetic vascular injury. Circulation 114 (6): 597-605. doi: 10.1161/CIRCULATIONAHA.106.621854

Goh SY, Cooper ME (2008) Clinical review: The role of advanced glycation end products in progression and complications of diabetes. The Journal of Clinical Endocrinology and Metabolism 93 (4): 1143-1152. doi: 10.1210/jc.2007-1817.

Reddy S, Bichler J, Wells-Knecht KJ et al. (1995) N epsilon-(carboxymethyl)lysine is a dominant advanced glycation end product (AGE) antigen in tissue proteins. Biochemistry 34 (34): 10872-10878.

Müller-Krebs S, Kihm LP, Madhusudhan T et al. (2012) Human RAGE antibody protects against AGE-mediated podocyte dysfunction. Nephrology Dialysis Transplantation 27 (8): 3129-3136. doi: 10.1093/ndt/gfs005.

Turk Z, Ljubic S, Turk N, Benko B (2001) Detection of autoantibodies against advanced glycation endproducts and AGE-immune complexes in serum of patients with diabetes mellitus. Clinica Chimica Acta 303 (1-2): 105-115.

Win MT, Yamamoto Y, Munesue S et al. (2012) Regulation of RAGE for attenuating progression of diabetic vascular complications. Experimental Diabetes Research 2012: 894605. doi: 10.1155/2012/894605.

Toyoda M, Suzuki D, Umezono T et al. (2004) Expression of human nephrin mRNA in diabetic nephropathy. Nephrology Dialysis Transplantation 19 (2): 380-385.

Vidotti D, Casarini DE, Cristovam PC et al. (2004) High glucose concentration stimulates intracellular renin activity and angiotensin II generation in rat mesangial cells. American Journal of Physiology-Renal Physiology 286 (6): F1039-F1045. Doi: 10.1152/ajprenal.00371.2003.

Teng B, Duong M, Tossidou I et al. (2014) Role of protein kinase C in podocytes and development of glomerular damage in diabetic nephropathy. Frontiers in Endocrinology 5: 179. doi: 10.3389/fendo.2014.00179.

Hoyer DP, Korkmaz Y, Grönke S et al. (2010) Differential expression of protein kinase C isoforms in coronary arteries of diabetic mice lacking the G-protein G11α. Cardiovascular Diabetology 29 (9): 93. doi: 10.1186/1475-2840-9-93.


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