Deneysel Olarak Oluşturulan Kontrast Madde Nefropatisinde Arginaz, Kreatinin ve Üre Üzerine Kalpain İnhibisyonunun Etkisi*

Abstract

Bu çalışmanın amacı, kontrast madde (KM) tarafından oluşturulan kontrast madde nefropatisinde (KMN), kalpain inhibisyonunu sağlayan

kalpain inhibitörü-1 (KAL-1)’in karaciğer arginaz aktivitesi ile serum üre ve kreatinin düzeyleri üzerine etkilerini araştırmaktır. Çalışmada, 28

adet 8-10 haftalık Sprague Dawley cinsi dişi ratlar kullanıldı. Deney hayvanları her grupta 7 hayvan olacak şekilde 4 gruba ayrıldı: Kontrol

grubu, KM verilen grup, KAL-1 verilen grup, KM ve KAL-1 verilen grup (KM+KAL-1). Kontrol grubuna herhangi bir uygulama yapılmadı.

Arginaz enzim aktivitesi Tiyosemicarbazid Diasetilmonoxim Üre (TDMU) metodu, serum kreatinin düzeyi Jaffe yöntemiyle, serum üre miktarı

otoanalizörde Kinetik UV Assay yöntemi ile yapıldı. Karaciğer arginaz enzim aktivitesi KM, KAL-1 ve KM+KAL-1 uygulanan ratlarda kontrol

grubuna göre düşük bulundu (p<0.001). Tek başına ve KM ile birlikte KAL-1 uygulaması, KM grubuna göre arginaz aktivitesinde daha fazla

düşüşe sebep olmuştur. Serum üre ve kreatininin değerleri KM grubunda, kontrol grubuna göre anlamlı olarak yüksek saptandı (p<0.01).

Serum kreatininin ve üre seviyesi KAL-1 ve KM+KAL-1 grubunda, KM grubuna göre önemli ölçüde düşüş gösterdi (p<0.01). Kreatinin ve üre

seviyesinin KM grubunda yüksek olması nefropatinin geliştiğini, KAL-1 verilen grupta bu değerlerin normale yaklaşması ise nefropatiyi KAL-

1’den kaynaklandığını düşündürmüştür. Arginaz enzim aktivitesinin ise KM grubunda önemli derecede düşmesi, KAL-1 uygulanmasının ise

bu düşüşü engellememesi; serum üre düzeyinde daha fazla artıştan korunmak için karaciğerin daha fazla üre üretmemesi amacı ile KAL-1’in

arginazın düşüşünü engellemediği şeklinde yorumlanabilir.

Keywords

• Arginaz,kalpain inhibitörü,kontrast madde nefropatisi,kreatinin,üre

Effects of Calpain Inhibition on Arginase, Creatinine and Urea in Experimental Contrast Medium- Induced Nephropathy

Abstract

The purpose of this study is to investigate the activity of liver arginase and examine the effects on the levels of serum urea and

creatinine of calpain inhibition (CAL-1) in contrast medium nephropathy (CMN) developed by contrast medium. Twenty-eight Sprague Dawley

female rats aged 8-10 weeks were used. The experimental animals were divided into 4 groups where there were animals in each namely

control, group given contrast medium (CM), group given CAL-1 (CAL-1) and group given CAL-1 and contract medium (CAL-1 + CM). No

application was made to the control group. Arginase enzyme activity was measured by Tiyosemicarbazid Diacetylmonoxim Urea (TDMU)

method. The level of serum creatinine was performed by Jaffe method and the amount of serum urea was performed by kinetic UV assay

method in autoanalyzer. Liver arginase enzyme activity in rats given CM, CAL-1, CM + CAL-1 was found significantly low compared to the rats

in the control group (p <0.001). The CAL-1 alone or in combination with CM application led to further declines in arginase activity compared to

the CM group. The levels of serum creatinine and urea in the CM group were found significant higher than that of control group (p <0.01). The

levels of serum creatinine and urea in CAL-1 and CM + CAL-1 showed a significantly decrease compared to the CM group (p <0.01). That the

levels of creatinine and urea are high in the CM group showed the development of nephropathy, and that these values being closer to normal

in the CAL-1 group showed that CAL-1 reduced nephropathy. The significant decrease in the activity of the arginase enzyme in the CM group,

and the CAL-1 application not preventing this decrease may be interpreted as CAL-1 did not prevent the decrease of arginase because more

urea was not produced in the liver to avoid further increases in the level of serum urea.

Keywords

• Arginase,calpain inhibitor,contrast medium nephropathy,creatinine,urea

References

1. 1. Briguori C, Tavano D, Colombo A. Contrast agent-associated nephrotoxicity. Prog Cardiovasc Dis 2003; 45: 493-503.
2. 2. Chatterjee PK, Brown PA, Cuzzocrea S, Zacharowski K, Stewart KN, Mota- Filipe H. Calpain inhibitor- 1 reduces renal ischemia/reperfusion injury in the rat. Kidney Int 2001; 59: 2073-83.
3. 3. Donkor IO. Calpain inhibitors: a survey of compounds reported in the patent and scientific literature. Expert Opin Ther Pat 2011; 21(5): 601-36.
4. 4. Dursun B, He Z, Somerset H, Oh DJ, Faubel S, Edelstein CL. Caspases and calpain are independent mediators of cisplatin- induced endothelial cell necrosis. Am J Physiol Renal Physiol 2006; 291: 578-87.
5. 5. Edelstein CL, Ling H, Gengaro PE, Nemenoff RA, Bahr BA, Schrier RW. Effect of glycine on prelethal and postlethal increases in calpain activity in rat renal proximal tubules. Kidney Int 1997; 52: 1271-8.
6. 6. Edelstein CL, Wieder ED, Yaqoob MM, Gengaro PE, Burke TJ, Nemenoff RA. The role of cysteine proteases in hypoxia- induced rat renal proximal tubular injury. Proc Natl Acad Sci USA 1995; 92: 7662-6.
7. 7. Edelstein CL, Shi Y, Schrier R. Role of caspases in hypoxia- induced necrosis of rat renal proximal tubules. J Am Soc Nephrol 1999; 10: 1940-9.
8. 8. Efron DT, Barbul A. Arginine and nutrition in renal disease. J Nutr 1999; 9(3): 142-4.

9. 9. Geyer JW, Dabich D. Rapid method for determination of arginase activity in tissue homogenates. Analy Biochem 1971; 39(2): 412-7.
10. 10. Jenkinson CP, Grigor MR. Rat mammary arginase: isolation and characterization. Biochem Med And Met Biol 1994; 51: 156-65.
11. 11. Kaysen GA, Strecter HJ. Purification and properties of arginase of rat kidney. Biochem J 1973; 133: 779-88.
12. 12. Katzberg RW, Haller C. Contrast-induced nephrotoxicity: clinical landscape. Kidney Int Suppl 2006: 3-7.
13. 13. Konarska L, Tomaszewski L, Rolczyk U. Studies on L-Arginase in developing rat small intestine, brain and kidney. Biochem Med Metab Biol 1986; 35: 170-8.
14. 14. Lindholt JS. Radiocontrast induced nephropathy. Eur J Vasc Endovasc Surg 2003; 25: 296-304.
15. 15. Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ. Protein measurements with the folin phenol reagent. J Biol Chem 1971; 193(1): 265-75.
16. 16. Parvez Z, Rahman MA, Moncada R. Contrast media-induced lipid peroxidation in the rat kidney. Invest Radiol 1989; 24: 697-702.
17. 17. Powers GS, Meister T. Urea synthesis and ammonia metabolism. Arias I, Popper H, Schachter D, Shafrits DA. eds. In: The Liver: Biology and Pathobiology. New York: Raven Press, 1982; pp 251-63.
18. 18. Reddan D, Laville M, Garovic VD. Contrast-induced nephropathy and its prevention: What do we really know from evidence-based findings? J Nephrol 2009; 22: 333-51.
19. 19. Romano G, Briguori C, Quintavalle C, Zanca C, Rivera NV, Colombo A. Contrast agents and renal cell apoptosis. Eur Heart J 2008; 29: 2569-76.
20. 20. Sorimachi H, Ishiura S, Suzuki K. Structure and physiological function of calpains. Biochem J 1997; 328: 721-32.