Advanced Glycation End Products (AGEs) Antibody Protects Against AGEs-induced Apoptosis and NF-ĸB p65 Subunit Overexpression in Rat Glomerular Culture

Oktavia Rahayu Adianingsih, Diana Lyrawati, Nur Samsu

Abstract


Advanced glycation end products (AGEs) have been thought to be a major cause of diabetic nephropathy (DN). The mechanisms underlying the involvement of AGEs antibody in diabetic nephropathy are not fully understood. The present study was designed to investigate the protective effect of AGEs antibody on AGEs-induced glomerular damage. Isolated glomeruli were pre-incubated either with 10 µg/mL polyclonal anti-AGEs antibody (AGE-pAb) or monoclonal anti-Nɜ -carboxymethyl-lysine antibody (CML-mAb) as a model of AGEs antibody to block interaction of AGEs with receptor for AGEs (RAGE) and incubated afterwards either with 100 µg/mL bovine serum albumin (BSA) or AGE-modified bovine serum albumin (AGE-BSA) for 48 h. Annexin V/nephrin doublestaining was performed to determine apoptosis. Using immunofluorescence, we found that administration of AGE-BSA not only significantly increased glomerular cells apoptosis and nuclear factor kappa B (NF-ĸB) p65 expression, but also reduced expression of nephrin, an important structural and signal molecule of podocytes slit diaphragm. Blocking the interaction of AGE-RAGE with AGEs antibody significantly protected glomerular cells from AGEs-induced apoptosis and NF-ĸB p65 overexpression. We found that AGE-pAb conferred superior protective effect compared with CmL-mAb for the same reduction in apoptosis and NF-ĸB p65 expression. In sharp contrast, CmL-mAb led to preserve expression of podocytes nephrin better than AGE-pAb. These results demonstrate that the antibody against AGEs may be beneficial for preventing the glomerular damage in DN.


Keywords


Advanced glycation end products, antibody, apoptosis, diabetic nephropathy, N e -(carboxymethyl)lysine

Full Text:

PDF

References


Guariguata L, Whiting DR, Hambleton I, Beagley J, Linnenkamp U, Shaw JE (2014) Global estimates of diabetes prevalence for 2013 and projections for 2035. Diabetes Res Clin Pract. 103(2):137–149.

Ott C, Jacobs K, Haucke E, Navarrete Santos A, Grune T, Simm A (2014) Role of advanced glycation end products in cellular signaling. Redox Biol. 2:411–429.

Yamagishi S-I, Matsui T (2010) Advanced glycation end products, oxidative stress and diabetic nephropathy. Oxid Med Cell Longev. 3(2):101–108.

Daroux M, Prévost G, Maillard-lefebvre H, Gaxatte C (2010) Advanced glycation end-products : Implications for diabetic and non-diabetic nephropathies. Diabetes Metab. 36:1–10.

Miyata T, Dan T (2008) Inhibition of advanced glycation end products (AGEs): An implicit goal in clinical medicine for the treatment of diabetic nephropathy ? Diabetes Res Clin Pract. 825:25–29.

Pashikanti S, Alba DR De, Boissonneault GA, Cervantes-laurean D (2010) Free Radical Biology & Medicine Rutin metabolites : Novel inhibitors of nonoxidative advanced glycation end products. Free Radic Biol Med. 48(5):656–663.

Joglekar MM, Panaskar SN, Arvindekar AU (2013) Inhibition of advanced glycation end product formation by cymene – A common food constituent. J Funct Foods. 6:107–115.

Losso JN, Bawadi HA, Chintalapati M (2011) Inhibition of the formation of advanced glycation end products by thymoquinone. Food Chem. 128(1):55–61.

Sang H, Gu J, Yuan J, Zhang M (2014) The protective effect of smilax glabra extract on advanced glycation end products-induced endothelial dysfunction in HUVECs via RAGE-ERK1/2-NF-κB pathway. J Ethnopharmacol. 155(1):785–795.

Feng L, Zhu M, Zhang M, Wang R, Tan X (2013) Protection of glycyrrhizic acid against AGEs-induced endothelial dysfunction through inhibiting RAGE/NF-κB pathway activation in human umbilical vein endothelial cells. J Ethnopharmacol. 148(1):27–36.

Reddy VP, Beyaz A (2006) Inhibitors of the Maillard reaction and AGE breakers as therapeutics for multiple diseases. Drug Discov Today. 11:646–654.

Mashitah MW, Azizah N, Samsu N, Indra MR, Bilal M, Yunisa MV, Arisanti AD (2015) Immunization of AGE-modified albumin inhibits diabetic nephropathy progression in diabetic mice. Diabetes, Metab Syndr Obes Targets Ther. 8:347–355.

Mera K, Nagai R, Takeo K, Izumi M, Maruyama T, Otagiri M (2011) An autoantibody against N-(carboxyethyl) lysine (CEL): Possible involvement in the removal of CEL-modified proteins by macrophages. Biochem Biophys Res Commun. 407(2):420–425.

Turk Z, Ljubic S, Turk N, Bojan B (2001) Detection of autoantibodies against advanced glycation endproducts and AGE-immune complexes in serum of patients with diabetes mellitus. Clin Chim Acta. 303:105–115.

Baydanoff S, Konova E, Ivanova (1996) N Determination of anti-AGE antibodies in human serum. Glycoconj J. 13:335–339.

Katsuya K, Yaoita E, Yoshida Y, Yamamoto Y, Yamamoto T (2006) An improved method for primary culture of rat podocytes. Int Soc Nephrol. 69:2101–2106.

Takemoto M, Asker N, Gerhardt H, Lundkvist A, Johansson BR, Saito Y, Betsholtz C (2002) Technical Advance A New Method for Large Scale Isolation of Kidney Glomeruli from Mice. Am J Pathol. 161(3):799–805.

Liu X, Fan Q, Yang G, Liu NAN, Chen D, Jiang YI, Wang L (2013) Isolating glomeruli from mice : A practical approach for beginners. Exp Ther Med. 5:1322–1326.

Mallipattu SK, Liu R, Zheng F et al (2012) Krüppel-like factor 15 (KLF15) is a key regulator of podocyte differentiation. J Biol Chem. 287(23):19122–19135.

Müller-Krebs S, Kihm LP, Madhusudhan T et al (2012) Human RAGE antibody protects against AGE-mediated podocyte dysfunction. Nephrol Dial Transplant. 27(8):3129–3136.

Bierhaus A, Schiekofer S, Schwaninger M et al (2001) Diabetes-associated sustained activation of the transcription factor nuclear factor-kappaB. Diabetes. 50(12):2792–2808.

Peng Y, Kim J, Park H et al (2016) AGE-RAGE signal generates a specific NF- κ B RelA “ barcode ” that directs collagen I expression. Nat Publ Gr. 1–10.

Liu R, Zhong Y, Li X et al (2014) Role of transcription factor acetylation in diabetic kidney disease. Diabetes. 63(7):2440–2453.

Fritz G (2011) RAGE: a single receptor fits multiple ligands. Trends Biochem Sci. 36(12):625–632.

Yao D, Brownlee M (2010) Hyperglycemia-induced reactive oxygen species increase expression of the receptor for advanced glycation end products (RAGE) and RAGE ligands. Diabetes. 59:249–255.

Fukami K, Ueda S, Yamagishi S et al (2004) AGEs activate mesangial TGF-beta-Smad signaling via an angiotensin II type I receptor interaction. Kidney Int. 66(6):2137–2147.

Ren Z, Liang W, Chen C et al (2012) Angiotensin II induces nephrin dephosphorylation and podocyte injury: role of caveolin-1. Cell Signal. 24(2):443–450.

Yu L, Lin Q, Feng J et al (2013) Inhibition of nephrin activation by c-mip through Csk–Cbp–Fyn axis plays a critical role in Angiotensin II-induced podocyte damage. Cell Signal. 25(3):581-588.

Tossidou I, Teng B, Menne J et al (2010) Podocytic PKC-alpha is regulated in murine and human diabetes and mediates nephrin endocytosis. PLoS One. 5(4):e10185.

Li X, Chuang PY, D’Agati VD et al (2015) Nephrin Preserves Podocyte Viability and Glomerular Structure and Function in Adult Kidneys. J Am Soc Nephrol. 1–17.

Hussain S, Romio L, Saleem M et al (2009) Nephrin Deficiency Activates NF-κB and Promotes Glomerular Injury. J Am Soc Nephrol. 20(8):1733–43.

Chuang PY, Dai Y, Liu R et al (2010) Alteration of forkhead box O (foxo4) acetylation mediates apoptosis of podocytes in diabetes mellitus. PLoS One. 6(8):e23566.

Ikezumi Y, Suzuki T, Karasawa T et al (2008) Activated macrophages down-regulate podocyte nephrin and podocin expression via stress-activated protein kinases. Biochem Biophys Res Commun. 376(4):706-711.

Kumar PA, Welsh GI, Raghu G et al (2016) Carboxymethyl lysine induces EMT in podocytes through transcription factor ZEB2: Implications for podocyte depletion and proteinuria in diabetes mellitus. Arch Biochem Biophys. 590:10–19.

Koito W, Araki T, Horiuchi S, Nagai R (2004) Conventional Antibody against Nε-(Carboxymethyl)Lysine (CML) Shows Cross-Reaction to Nε-(Carboxyethyl)Lysine (CEL): Immunochemical Quantification of CML with a Specific Antibody. J Biochem. 136(6):831–837.




DOI: http://dx.doi.org/10.11594/jtls.06.03.08

Copyright (c) 2016 Journal of Tropical Life Science