HEPARANASE EXPRESSION IN RENAL INTERSTITIAL MAY CONTRIBUTE TO EPITHELIAL AND ENDOTHELIAL CELLS INJURIES AFTER KIDNEY ISCHEMIC/ REPERFUSION EPISODE IN MICE
Kidney ischemia/reperfusion injury (I/R) is the most frequent cause of acute kidney injury (AKI). It had been reported that epithelial and endothelial injuries occurred during kidney I/R injury. Heparanase is an enzyme that degrades glycocalyx and contributes to I/R injury in the heart and liver. This study is to elucidate the association between heparanase expression and cell injuries in kidney I/R injury. We performed kidney I/R injury model in mice using renal pedicle clamping for 30 minutes and sacrificed the mice in day 1 (n=6) after operation. Sham-operation procedure (SO, n=5) was used as control. PAS staining was used to quantify tubular injury score. Serum creatinine was measured from orbital venous. Heparanase expression was quantified using western blot and real-time PCR. Heparanase localization and endothelial injury were shown by immunostaining of heparanase and double glycocalyx-von Willebrand factor. Kidney I/R induced an increase of serum creatinine level that was accompanied by elevation of tubular injury score and glycocalyx damage. Glycocalyx damage was identified using immunofluorescent staining that revealed a disruption of glycocalyx or lectin layer in the endothelial cells of intra-renal artery. This finding was associated with significant elevation of heparanase mRNA and protein level expression. We found that heparanase was expressed in the renal epithelial and interstitial cells. In conclusion, heparanase may induce endothelial and epithelial injury in the kidney I/R episode. Using heparanase expression as a early marker of AKI may possibly promising.
Keywords: kidney I/R, epithelial injury, endothelial injury, heparanase, glycocalyx
Arfian, N., N. Emoto, N. Vignon-Zellweger, and K. Nakayama. 2012. ET-1 deletion from endothelial cells protects the kidney during the extension phase of ischemia/reperfusion injury. Biochem Biophys Res Commun 425: 443-449.
Beresewicz, A., E. Czarnowska, and M. Maczewski. 1998. Ischemic preconditioning and superoxide dismutase protect against endothelial dysfunction and endothelium glycocalyx disruption in the postischemic guinea-pig hearts. Mol Cell Biochem 186: 87-97.
Bonventre, J.V., and L. Yang. 2011. Cellular pathophysiology of ischemic acute kidney injury. J Clin Invest 121: 4210-4221.
Brodsky, S.V., T. Yamamoto, T. Tada, and B. Kim. 2002. Endothelial dysfunction in ischemic acute renal failure: rescue by transplanted endothelial cells. Am J Physiol Renal Physiol 282: F1140-1149.
Inazaki, K., Y. Kanamaru, Y. Kojima, and N. Sueyoshi. 2004. Smad3 deficiency attenuates renal fibrosis, inflammation,and apoptosis after unilateral ureteral obstruction. Kidney Int 66: 597-604.
Jeansson, M., K. Bjorck, O. Tenstad, and B. Haraldsson. 2009. Adriamycin alters glomerular endothelium to induce proteinuria. J Am Soc Nephrol 20: 114-122.
Kim, J., K.J. Jung, and K.M. Park. 2010. Reactive oxygen species differently regulate renal tubular epithelial and interstitial cell proliferation after ischemia and reperfusion injury. Am J Physiol Renal Physiol 298: F1118-1129.
Kisanuki, Y.Y., N. Emoto, T. Ohuchi, and B. Widyantoro. 2011. Low blood pressure in endothelial cell-specific endothelin 1 knockout mice. Hypertension 56: 121-128.
Kramer, A., M. van den Hoven, A. Rops, and T. Wijnhoven. 2006. Induction of glomerular heparanase expression in rats with adriamycin nephropathy is regulated by reactive oxygen species and the renin-angiotensin system. J Am Soc Nephrol 17: 2513-2520.
Li, J., J.P. Li, X. Zhang, Z. Lu. 2012. Expression of heparanase in vascular cells and astrocytes of the mouse brain after focal cerebral ischemia. Brain Res 1433: 137-144.
Lipowsky, H.H., L. Gao, and A. Lescanic. 2011. Shedding of the endothelial glycocalyx in arterioles, capillaries, and venules and its effect on capillary hemodynamics during inflammation. Am J Physiol Heart Circ Physiol 301: H2235-2245.
Maxhimer, J.B., M. Somenek, G. Rao, C.E. Pesce. 2005. Heparanase-1 gene expression and regulation by high glucose in renal epithelial cells: a potential role in the pathogenesis of proteinuria in diabetic patients. Diabetes 54: 2172-2178.
Molitoris, B.A., R. Sandoval, and T.A. Sutton. 2002. Endothelial injury and dysfunction in ischemic acute renal failure. Crit Care Med 30: S235-240.
Molitoris, B.A., and T.A. Sutton. 2004. Endothelial injury and dysfunction: role in the extension phase of acute renal failure. Kidney Int 66: 496-499
Mulivor, A.W., and H.H. Lipowsky. 2004. Inflammation-and ischemia-induced shedding of venular glycocalyx. Am J Physiol Heart Circ Physiol 286: H1672-1680.
Rao, G., H.G. Ding, W. Huang, and D. Le. 2011. Reactive oxygen species mediate high glucose-induced heparanase-1 production and heparan sulphate proteoglycan degradation in human and rat endothelial cells: a potential role in the pathogenesis of atherosclerosis. Diabetologia 54: 1527-1538.
Rehm, M., D. Bruegger, F. Christ, and P. Conzen. 2007. Shedding of the endothelial glycocalyx in patients undergoing major vascular surgery with global and regional ischemia. Circulation 116: 1896-1906.
Salmon, A.H., and S.C. Satchell. 2012. Endothelial glycocalyx dysfunction in disease: albuminuria and increased microvascular permeability. J Pathol 226: 562-574.
Schrier R.W., W. Wang, B. Poole, and A. Mitra. 2004. Acute renal failure: definitions, diagnosis, pathogenesis, and therapy. J Clin Invest 114: 5-14.
Singh, A., S.C. Satchell, C.R. Neal, E.A. McKenzie. 2007. Glomerular endothelial glycocalyx constitutes a barrier to protein permeability. J Am Soc Nephrol 18: 2885-2893.
Sutton, T.A., C.J. Fisher, and B.A. Molitoris. 2002. Microvascular endothelial injury and dysfunction during ischemic acute renal failure. Kidney Int 62: 1539-1549.
Sutton, T.A., H.E. Mang, S.B. Campos, and R.M. Sandoval. 2003. Injury of the renal microvascular endothelium alters barrier function after ischemia. Am J Physiol Renal Physiol 285: F191-198.
van den Hoven, M.J., A.L. Rops, I. Vlodavsky, V. Levidiotis. 2007. Heparanase in glomerular diseases. Kidney Int 72: 543-548.
van den Hoven, M.J., F. Waanders, A.L. Rops, and A.B. Kramer AB. 2009. Regulation of glomerular heparanase expression by aldosterone, angiotensin II and reactive oxygen species. Nephrol Dial Transplant 2009; 24: 2637-2645.
Wu, H., G. Chen, K.R. Wyburn, and J. Yin. 2007. TLR4 activation mediates kidney ischemia/ reperfusion injury. J Clin Invest 117: 2847-2859. 9