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Saudi Journal of Kidney Diseases and Transplantation
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ORIGINAL ARTICLE Table of Contents   
Year : 2009  |  Volume : 20  |  Issue : 4  |  Page : 604-607
Serum trypsin inhibitory capacity in hemodialysis patients

1 Department of Clinical Biochemistry, Zahedan University of Medical Sciences, Zahedan, Iran
2 Research Center for Infectious Diseases and Tropical Medicine, Bou Ali Hospital, Zahedan University of Medical Sciences, Zahedan, Iran
3 Department of Nutrition, School of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran
4 Manitoba Institute of Cell Biology and Department of Biochemistry and Medical Genetics, University of Manitoba, Winnipeg, Canada

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Date of Web Publication8-Jul-2009


It has been established that overproduction of reactive oxygen species (ROS) occurs during hemodialysis causing oxidation of proteins. Alpha-1-antitrypsin is the major circulating anti­protease which contains methionine in the active site. The aim of the present study was to measure the level of serum trypsin inhibitory capacity (sTIC) in hemodialysis patients. This case-control study was performed in 52 hemodialysis patients and 49 healthy controls. sTIC was measured by enzymatic assay. The sTIC was significantly (P< 0.001) lower in hemodialysis patients (1.87 ± 0.67 µmol/min/mL) than healthy controls (2.83 ± 0.44 µmol/min/L). Reduction of sTIC may be due to the oxidation of methionine residue in the reactive site of alpha-1 antitrypsin.

Keywords: Alpha-1 Anitrypsin, Hemodialysis, Chronic renal failure, Serum trypsin inhibitory capacity

How to cite this article:
Hashemi M, Mehrabifar H, Homayooni F, Naderi M, Montazerifar F, Ghavami S. Serum trypsin inhibitory capacity in hemodialysis patients. Saudi J Kidney Dis Transpl 2009;20:604-7

How to cite this URL:
Hashemi M, Mehrabifar H, Homayooni F, Naderi M, Montazerifar F, Ghavami S. Serum trypsin inhibitory capacity in hemodialysis patients. Saudi J Kidney Dis Transpl [serial online] 2009 [cited 2021 Jan 16];20:604-7. Available from: https://www.sjkdt.org/text.asp?2009/20/4/604/53248

   Introduction Top

Hemodialysis (HD) is the commonest renal replacement therapy used to treat end stage renal disease (ESRD). The presence of a chro­nic inflammatory status has been documented in end-stage renal disease patients receiving maintenance HD. [1] ,[2] HD reduces serum antioxi­dative capacity [3] and susceptibility to oxidative stress in patients is mediated by lower antio­xidant defenses. Previous studies have demons­trated several deficiencies in hydrophilic antio­xidant molecules in plasma of HD patients with a lower concentration of vitamin C. [4] It has been reported that superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities were lo­wer in erythrocytes isolated from HD patients. [5] Some reports showed that oxidation of protein also occur in hemodialysis patients. [6],[7],[8],[9]

Alpha-1 antitrypsin (AAT) is a serine protease inhibitor which complexes with the active site of serine proteases, thus blocking their enzyme activity. AAT has a single polypeptide chain of 394 amino acid residues with three carbohydrate side chains, giving a total molecular weight of 51 kDa. Methionine is at the reactive site of hu­man alpha-1-antitrypsin and can be oxidized readily with peroxide, hydroxyl radicals, hypo­chlorite, chloramines, and peroxynitrite anions. [10],[11],[12],[13] Ueda et al reported higher serum concentra­tions of oxidized AAT in patients with infla­mmatory and rheumatoid diseases than healthy subjects concluding that oxidized AAT can be a marker of oxidative stress. [14]

The aim of the present study was to determine the level of serum trypsin inhibitory capacity (sTIC) in hemodialysis patients.

   Material and Methods Top


Trypsin, a-N-benzyl-DL-arginine-p-nitroaniline (BAPNA), bovine serum albumin (BSA) were obtained from Sigma. Tris (hydroxylmethyl)­aminomethane (Tris base), CaCl2, HCl, Dimethyl sulfoxide (DMSO) and acetic acid were ana­lytical grade and were obtained from Merck chemical company


This study was performed between May 2007 to September 2007 in hemodialysis clinic of Khatam Al Anbia hospital, Zahedan, Iran. The project was approved by ethical committee of Zahedan University of Medical Sciences and con­sent was obtained from all patients and healthy individuals. Blood samples were obtained from 52 hemodialysis patients and 49 healthy controls. The serum was separated and stored at -80 °C until analyzed.

s-TIC assay

s-TIC was measured using enzymatic assay described earlier. [13],[14],[15]

Statistical analysis:

All values are given as mean ± standard de­viation. Student t test was used to compare the significance between means and P value of < 0.05 was taken as significant.

   Results Top

Total of 52 hemodialysis (31 male, 21 female, age range 19-80 years), and 49 healthy control subjects (35 male, 14 female, age range 22-60 years) were included in the study. The mean duration of dialysis was 42.48 ± 41.68 months. The serum trypsin inhibitory capacity (sTIC) was determined in cases and controls. As shown in [Figure 1], the level of sTIC was significantly lower (P< 0.001) in hemodialysis patients (1.87 ± 0.67 µmol/min/mL) than in normal subjects (2.83 ± 0.44 µmol/min/mL).

   Discussion Top

Patients with end stage renal disease on main­tenance hemodialysis are known to have in­creased levels of oxidative stress and produc­tion of free radicals resulting in imbalance bet­ween oxidant and antioxidant systems. [16],[17],[18] Oxi­dation of proteins was infact documented to occur in hemodialysis patients. [6],[7],[8],[9] In the present study, we investigated the possible changes in serum trypsin inhibitory capacity in hemodia­lysis patients. The results confirm that serum trypsin inhibitory capacity was significantly lo­wer in hemodialysis patients than normal sub­jects.

Borawski et al on the other hand found higher serum AAT levels in patients with elevated CRP and hypoalbuminemia suggesting that it may be the marker of severe inflammation in hemodialysis patients. [19]

Docci et al found significantly lower concen­trations of AAT in patients with dialysis related arthropathy suggesting a role in amyloidoge­nesis. [20] Cristea et al also reported that alpha-1 antitrypsin decrease during hemodialysis. [21]

It has been shown that exposure of humans to nitrite resulted in significant inactivation of alpha 1-protease inhibitor (alpha 1-PI) in the broncho­alveolar lavage fluid suggesting that oxidation of methionine residue occurs at the active site of this protein. [22] Ueda et al reported that the serum concentrations of oxidized AAT in pa­tients with inflammatory and rheumatoid di­seases were higher than healthy subjects. [14] Mo­raga et al reported that oxidized AAT activated monocytes, which in turn activated NADPH oxidase, and production of chemokinines and cytokines. [23] They concluded that oxidized AAT is not only a product of inflammatory reaction, but also plays a role in enhancing inflammatory reaction.

The oxidation of methionine residues in the reactive site of AAT to methionine sulfoxide can result in functional inactivation of anti­protease activity of AAT. [24],[25],[26],[27] From all this data it is evident that hemodialysis and uremic toxins activate leukocytes leading to overproduction of reactive oxygen species that increases oxidation of methionine residues of AAT resulting in de­crease inhibitory activity against proteases. In conlusion, the serum trypsin inhibitorty capa­city (sTIC) may be used as a marker of oxi­dative stress in hemodialysis patients.

   Acknowledgments Top

This work was supported by a research grant from Zahedan University of Medical Sciences. The authors would like to thank all the patients and subjects who participated in the study.

   References Top

1.Coskun C, Kural A, Doventas Y, et al. Hemo­dialysis and protein oxidation products. Ann N Y Acad Sci 2007;1100:404-8.  Back to cited text no. 1    
2.Valentini J, Grotto D, Paniz C, et al. The influence of the hemodialysis treatment time under oxidative stress biomarkers in chronic renal failure patients. Biomed Pharmacother 2008;62(6):378-82.  Back to cited text no. 2    
3.Nakayama K, Terawaki H, Nakayama M, et al. Reduction of serum ant oxidative capacity during hemodialysis. Clin Exp Nephrol 2007;11 (3):218-24.  Back to cited text no. 3    
4.Bakaev VV, Efremov AV, Tityaev, II. Low levels of dehydroascorbic acid in uraemic serum and the partial correction of dehydroascorbic acid deficiency by haemodialysis. Nephrol Dial Transplant 1999;14(6):1472-4.  Back to cited text no. 4    
5.Paul JL, Sall ND, Soni T, et al. Lipid peroxi­dation abnormalities in hemodialyzed patients. Nephron 1993;64(1):106-9.  Back to cited text no. 5    
6.Witko Sarsat V, Friedlander M, Capeillere­Blandin C, et al. Advanced oxidation protein products as a novel marker of oxidative stress in uremia. Kidney int 1996;49(5):1304-13.  Back to cited text no. 6    
7.Mera K, Anraku M, Kitamura K, et al. Oxida­tion and carboxy methyl lysine modification of albumin: possible involvement in the progre­ssion of oxidative stress in hemodialysis patients. Hypertens Res 2005;28(12):973-80.  Back to cited text no. 7    
8.Sutherland WH, Gieseg SP, Walker RJ, et al. Serum protein-bound 3,4-dihydroxyphenylalanine and related products of protein oxidation and chronic hemodialysis. Ren Fail 2003;25(6):997- 1009.  Back to cited text no. 8    
9.Mayer B, Zitta S, Greilberger J, et al. Effect of hemodialysis on the antioxidative properties of serum. Biochim Biophys Acta 2003;1638(3): 267-72.  Back to cited text no. 9    
10.Ghavami S, Hashemi M, de Serres FJ, et al. Trypsin inhibitory capacity in vernal keratoc­onjunctivitis. Invest Ophthalmol Vis Sci 2007; 48(1):264-9.  Back to cited text no. 10    
11.Ghavami S, Hashemi M, Shahriari HA, et al. Alpha-1-antitrypsin phenotypes and HLA-B27 typing in uveitis patients in southeast Iran. Clin Biochem 2005;38(5):425-32.  Back to cited text no. 11    
12.Hashemi M, Alavian SM, Ghavami S, et al. High prevalence of alpha 1 antitrypsin pheno­types in viral hepatitis B infected patients in Iran. Hepatol Res 2005;33(4):292-7.  Back to cited text no. 12    
13.Hashemi M, Naderi M, Rashidi H, Ghavami S. Impaired activity of serum alpha-1-antitrypsin in diabetes mellitus. Diabetes Res Clin Pract 2007;75(2):246-8.  Back to cited text no. 13    
14.Ueda M, Mashiba S, Uchida K. Evaluation of oxidized alpha-1-antitrypsin in blood as an oxidative stress marker using anti oxidative alpha1-AT monoclonal antibody. Clin Chim Acta 2002; 317(1-2):125-31.  Back to cited text no. 14    
15.Dietz AA, Rubinstein HM, Hodges LK. Mea­surement of alpha 1 antitrypsin in serum, by immunodiffusion and enzymatic assay. Clin Chem Lab Med 1974;20:396-9.  Back to cited text no. 15    
16.Pawlak K, Pawlak D, Mysliwiec M. Impaired renal function and duration of dialysis therapy are associated with oxidative stress and pro­atherogenic cytokine levels in patients with end­stage renal disease. Clin Biochem 2007;40(1­2):81-5.  Back to cited text no. 16    
17.Ferretti G, Bacchetti T, Masciangelo S, Pallotta G. Lipid peroxidation in hemodialysis patients: Effect of vitamin C supplementation. Clin Biochem 2008;41(6):381-6.  Back to cited text no. 17    
18.Erdogan C, Unlucerci Y, Turkmen A, et al. The evaluation of oxidative stress in patients with chronic renal failure. Clin Chim Acta 2002; 322(1-2):157-61.  Back to cited text no. 18    
19.Borawski J, Naumnik B, Mysliwiec M. Serum alpha1antitrypsin but not complement C3 and C4 predicts chronic inflammation in hemo­dialysis patients. Ren Fail 2003;25(4):589-93.  Back to cited text no. 19    
20.Docci D, Bilancioni R, Baldrati L, et al. Serum alpha-1-antitrypsin in hemodialysis patients with dialysis arthropathy. Int J Artif Organs 1993;16(3):123-7.  Back to cited text no. 20    
21.Cristea A, Rus HG, Gherman M, Manasia M. Decrease of alpha 1-antitrypsin during hemo­dialysis. Med Interne 1981;19(4):379-81.  Back to cited text no. 21    
22.Mohsenin V, Gee JL. Oxidation of alpha 1­protease inhibitor: role of lipid peroxidation products. J Appl Physiol 1989;66(5):2211-5.  Back to cited text no. 22    
23.Moraga F, Janciauskiene S. Activation of primary human monocytes by the oxidized form of alpha 1-antitrypsin. J Biol Chem 2000;275 (11):7693-700.  Back to cited text no. 23    
24.Carp H, Janoff A. Potential mediator of in­flammation. Phagocyte-derived oxidants suppress the elastase inhibitory capacity of alpha 1­proteinase inhibitor in vitro. J clin Invest 1980; 66(5):987-95.  Back to cited text no. 24    
25.Carp H, Janoff A. Inactivation of bronchial mucous proteinase inhibitor by cigarette smoke and phagocyte-derived oxidants. Exp Lung Research 1980;1(3):225-37.  Back to cited text no. 25    
26.Matheson NR, Wong PS, Travis J. Enzymatic inactivation of human alpha-1-proteinase inhi­bitor by neutrophil myeloperoxidase. Biochem Biophys Research Communications 1979;88(2) :402-9.  Back to cited text no. 26    
27.Swaim MW, Pizzo SV. Methionine sulfoxide and the oxidative regulation of plasma pro­teinase inhibitors. J Leukocyte biol. 1988;43(4): 365-79.  Back to cited text no. 27    

Correspondence Address:
Mohammad Hashemi
Department of Clinical Biochemistry, School of Medicine, Zahedan University of Medical Sciences, Zahedan, I.R.
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PMID: 19587500

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