| |
|
|
| Year : 2009 | Volume
: 20
| Issue : 4 | Page : 604-607 |
|
| Serum trypsin inhibitory capacity in hemodialysis patients |
|
|
Mohammad Hashemi1, Hamid Mehrabifar2, Fatemeh Homayooni2, Mohammad Naderi2, Farzaneh Montazerifar3, Saeid Ghavami4
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
Click here for correspondence address and email
| Date of Web Publication | 8-Jul-2009 |
|
|
 |
|
 Abstract | | |
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 | |  |
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 | | |
Materials
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
Patients
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 | | |
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 | | |
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 | | |
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 | | |
| 1. | Coskun C, Kural A, Doventas Y, et al. Hemodialysis and protein oxidation products. Ann N Y Acad Sci 2007;1100:404-8.  |
| 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. |
| 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. |
| 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. |
| 5. | Paul JL, Sall ND, Soni T, et al. Lipid peroxidation abnormalities in hemodialyzed patients. Nephron 1993;64(1):106-9. |
| 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. |
| 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. |
| 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. |
| 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. |
| 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. |
| 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. |
| 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. |
| 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. |
| 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. |
| 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. |
| 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. |
| 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. |
| 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. |
| 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. |
| 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. |
| 21. | Cristea A, Rus HG, Gherman M, Manasia M. Decrease of alpha 1-antitrypsin during hemodialysis. Med Interne 1981;19(4):379-81. |
| 22. | Mohsenin V, Gee JL. Oxidation of alpha 1protease inhibitor: role of lipid peroxidation products. J Appl Physiol 1989;66(5):2211-5. |
| 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. |
| 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. |
| 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. |
| 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. |
| 27. | Swaim MW, Pizzo SV. Methionine sulfoxide and the oxidative regulation of plasma proteinase inhibitors. J Leukocyte biol. 1988;43(4): 365-79. |
Correspondence Address:
Mohammad Hashemi
Department of Clinical Biochemistry, School of Medicine, Zahedan University of Medical Sciences, Zahedan, I.R.
Iran
  | Check |
PMID: 19587500 
[Figure 1] |
|
| This article has been cited by | | 1 |
Alpha-1 antitrypsin: Itęs role in health and disease |
|
| Hashemi, M. and Sharma, P. and Eshraghi, M. and Naderi, M. and Moazeni-Roodi, A. and Mehrabifar, H. and Taheri, M. | | Anti-Inflammatory and Anti-Allergy Agents in Medicinal Chemistry. 2010; 9(4): 279-288 | | [Pubmed] | |
|
|
|
|
|
|
|
|
|
|
|
|
| Article Access Statistics | | | Viewed | 2570 | | | Printed | 76 | | | Emailed | 0 | | | PDF Downloaded | 441 | | | Comments | [Add] | | | Cited by others | 1 | | |
|
|
