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* Department of Medicine, Harbor–UCLA Medical Center, UCLA School of Medicine, Torrance, CA 90502 and Wakunaga of America Co., Ltd., Mission Viejo, CA 92691
4 To whom correspondence should be addressed. E-mail: ysniihara{at}msn.com.
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ABSTRACT |
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KEY WORDS: • aged garlic extract • sickle-cell anemia • S-allylcysteine
Sickle-cell disease is one of the most prevalent hereditary disorders with prominent morbidity and mortality. The disease may affect various ethnic groups, such as people of Hispanic and Middle Eastern descent, but it affects people of African descent the most. Clinical manifestations of sickle-cell disease are largely due to a hemolytic process leading to severe anemia and vaso-occlusion, resulting in pain and organ damage. In the pathophysiology of sickle-cell disease, increased oxidant susceptibility of sickle red blood cells (RBC)5 has been demonstrated to play a major role (1–7).
Recent investigations have brought forth ample data that support significant antioxidant activity of garlic (Allium sativum) (8–12). Among various preparations of garlic supplements, AGE, in particular, has been associated with antioxidant activities in sound scientific experiments (8,10,11,13–18). Therefore, based on these data, we have examined the potential role of AGE as an antioxidant in sickle-cell disease.
AGE. AGE (Kyolic), donated by Wakunaga of America, is formulated by soaking sliced raw garlic in 15 to 20% aqueous ethanol for up to 20 mo at room temperature. The extract is then filtered and concentrated under reduced pressure at low temperature. The content of water-soluble compounds is relatively high, whereas that of oil-soluble compounds is low. The AGE used in this trial contained 305 g/L of extracted solids; S-allylcysteine, the most abundant water-soluble organosulfur compound in AGE, was present at a concentration of 1.47 g/L.
Study population. The participants were individuals with an established diagnosis of sickle-cell anemia [hemoglobin (HGB)SS] by hemoglobin electrophoresis and were 18 y or more of age. The exclusion criteria were any significant medical conditions other than sickle-cell disease, including diabetes mellitus, renal failure, or heart failure, pregnancy, and history of treatment with any antisickling agents within 12 mo of the initiation of the study. This project was approved by the Internal Review Board of Harbor–UCLA Research and Education Institute. All participants were volunteers and signed appropriate consent forms after careful explanation and review of the protocol.
Administration of AGE. After obtaining consent, each patient was seen at baseline for interview, physical examination, and baseline blood tests. A urine pregnancy test was also performed for each woman of child-bearing age. Participants were instructed to self-administer liquid AGE at a dose of 5 mL daily.
At 4 wk, the patients were reevaluated with a brief physical examination and interview. Also, whole blood samples were drawn for evaluation, including the Heinz body test.
Biochemical and physiological parameters. A Coulter counter was used for determination of RBC counts and HGB levels.
Heinz bodies were evaluated with a standard method using crystal violet solution (19–21). The number of RBCs containing five or more Heinz bodies was counted and expressed as a percentage.
Statistical analysis. All values are reported as means ± SD The paired Student's t test was used to evaluate the differences of variables between baseline and follow-up data. All tests of significance were two-tailed, and significance was defined at P < 0.05.
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DISCUSSION |
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TOPABSTRACTRESULTSDISCUSSIONLITERATURE CITED |
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The Heinz bodies, which adhere to the RBC membrane (26), may themselves cause significant damage to the membrane. In any case, assessment of Heinz bodies is a useful gauge in evaluating susceptibility of RBC to oxidant stress (4,7). The data in the study are preliminary in nature. However, AGE therapy was associated with a decrease in Heinz bodies in sickle RBC in each patient. The data were consistent with our hypothesis and confirm previous reports that demonstrated antioxidant activities of AGE (8,10,11,13–18).
In regard to hematological parameters, AGE had no significant effect on RBC count, HGB level, and HCT. Reports (27–32) have shown in animals that garlic extract may affect the RBC count adversely during the acute phase by inducing hemolytic anemia. Ironically, these are thought to be caused, at least partially, by oxidant stress caused by compounds such as allicin contained in ordinary garlic extract. Allicin has been shown to enhance LDL oxidation (16) and to oxidize the iron of HGB in RBC with methemoglobin formation (33). AGE, on the other hand, has been processed to eliminate these compounds without the loss of water-soluble antioxidant compounds such as S-allylcysteine and fructosyl arginine. One of these water-soluble compounds, S-allylcysteine, has been shown in vitro to inhibit the formation of dense cells in blood samples from sickle-cell anemia patients (34,35). Thus, AGE appears to have less capacity for inducing oxidant stress than ordinary garlic extracts (36), yet it maintains its antioxidant activity (8,10,11,13–18). In this study, there were no adverse effects, including hematological parameters, during the few weeks of AGE administration.
In conclusion, we have demonstrated, in a small cohort of sickle-cell anemia patients, an association of AGE therapy with a decrease in Heinz bodies in their RBCs. One must be cautioned that the data presented here are preliminary and the study was an open-labeled nonrandomized trial. However, the results suggest that AGE has a potential effect as an antioxidant in sickle-cell disease. Previously, AGE has been shown to significantly improve erythrocyte deformability through stabilization of erythrocyte membranes in nonsickle RBCs (37). These phenomena were attributed to the antioxidant activities of AGE (37). At baseline, erythrocyte deformability is further altered in sickle RBC due to an abnormal, highly permeable membrane (38). This abnormality is thought to contribute, at least partially, to the formation of dense cells (39). The dense cells, in turn, react with inflammatory cells and endothelial cells leading to vaso-occlusive changes (40). AGE may also improve erythrocyte membrane stability in sickle RBC through antioxidant activities, as suggested by the data presented here in which there was a reduction of Heinz bodies in sickle RBC during daily AGE administration. Again, the data are preliminary, but they are consistent with previous findings regarding antioxidant effects of AGE. Further testing is warranted for confirmation of the efficacy of this relatively harmless agent in the management of sickle-cell disease.
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FOOTNOTES |
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2 Author disclosure: Harunobu Amagase is employed by Wakunaga of America, Ltd.
3 Supported by a grant from the National Heart, Lung, and Blood Institute, National Institutes of Health; contract grant no. 5R29HL58640–01.
5 Abbreviations used: GSH, reduced glutathione; HGB, hemoglobin; HCT, hematocrit; NAD, nicotinamide adenine dinucleotide; RBC, red blood cell; RET, reticulocyte.
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LITERATURE CITED |
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1. Chiu D, Lubin B. Abnormal vitamin E and glutathione peroxidase levels in sickle cell anemia: evidence for increased susceptibility to lipid peroxidation in vivo. J Lab Clin Med. 1979;94:542–8.[Medline]
2. Hebbel RP, Eaton JW, Balasingam M, Steinberg MH. Spontaneous oxygen radical generation by sickle erythrocytes. J Clin Invest. 1982;70:1253–9.[Medline]
3. Jain SK, Shohet SB. A novel phospholipid in irreversibly sickled cells: evidence for in vivo peroxidative membrane damage in sickle cell disease. Blood. 1984;63:362–7.
4. Lachant NA, Davidson WD, Tanaka KR. Impaired pentose phosphate shunt function in sickle cell disease: a potential mechanism for increased Heinz body formation and membrane lipid peroxidation. Am J Hematol. 1983;15:1–13.[Medline]
5. Rank BH, Carlsson J, Hebbel RP. Abnormal redox status of membrane-protein thiols in sickle erythrocytes. J Clin Invest. 1985;75:1531–7.[Medline]
6. Rice-Evans C, Omorphos SC, Baysal E. Sickle cell membranes and oxidative damage. Biochem J. 1986;237:265–9.[Medline]
7. Wetterstroem N, Brewer GJ, Warth JA, Mitchinson A, Near K. Relationship of glutathione levels and Heinz body formation to irreversibly sickled cells in sickle cell anemia. J Lab Clin Med. 1984;103:589–96.[Medline]
8. Balasenthil S, Arivazhagan S, Nagini S. Garlic enhances circulatory antioxidants during 7, 12-dimethylbenz[a]anthracene-induced hamster buccal pouch carcinogenesis. J Ethnopharmacol. 2000;72:429–33.[Medline]
9. Grudzinski IP, Frankiewicz-Jozko A, Bany J. Diallyl sulfide—a flavour component from garlic (Allium sativum) attenuates lipid peroxidation in mice infected with Trichinella spiralis. Phytomedicine. 2001;8:174–7.[Medline]
10. Ide N, Lau BH. Garlic compounds protect vascular endothelial cells from oxidized low density lipoprotein-induced injury. J Pharm Pharmacol. 1997;49:908–11.[Medline]
11. Ide N, Lau BH. Garlic compounds minimize intracellular oxidative stress and inhibit nuclear factor-
B activation. J Nutr. 2001;131:1020S–106S.
12. Wu CC, Sheen LY, Chen HW, Tsai SJ, Lii CK. Effects of organosulfur compounds from garlic oil on the antioxidation system in rat liver and red blood cells. Food Chem Toxicol. 2001;39:563–9.[Medline]
13. Ho SE, Ide N, Lau BH. S-allyl cysteine reduces oxidant load in cells involved in the atherogenic process. Phytomedicine (Jena). 2001;8:39–46.
14. Ide N, Lau BH. Aged garlic extract attenuates intracellular oxidative stress. Phytomedicine (Jena). 1999;6:125–31.
15. Ide N, Lau BH. S-allylcysteine attenuates oxidative stress in endothelial cells. Drug Dev Ind Pharm. 1999;25:619–24.[Medline]
16. Ide N, Lau BHS, Ryu K, Matsuura H, Itakura Y. Antioxidant effects of fructosyl arginine, a Maillard reaction product in aged garlic extract. J Nutr Biochem. 1999;10:372–6.[Medline]
17. Ide N, Nelson AB, Lau BH. Aged garlic extract and its constituents inhibit Cu(2+)-induced oxidative modification of low density lipoprotein. Planta Med. 1997;63:263–4.[Medline]
18. Ryu K, Ide N, Matsuura H, Itakura Y. N
-(1-Deoxy-D-fructos-1-yl)-L-arginine, an antioxidant compound identified in aged garlic extract. J Nutr. 2001;131:972S–6S.
19. Beutler E, Dern RJ, Alving AS. The hemolytic effect of primaquine: VI. An in vitro test for sensitivity of erythrocytes to primaquine. J Lab Clin Med. 1955;45:40–50.[Medline]
20. Henry JB. Clinical diagnosis and management by laboratory methods. Philadelphia: WB Saunders; 1979.
21. Pearce CJ, Dow P. Anemia of abnormal globin development—Hemoglobinopathies. Philadelphia: JB Lippincott; 1990. p. 185–211.
22. Asakura T, Onishi T, Friedman S, Schwartz E. Abnormal precipitation of oxyhemoglobin S by mechanical shaking. Proc Natl Acad Sci USA. 1974;71:1594–8.
23. Das SK, Nair RC. Superoxide dismutase, glutathione peroxidase, catalase and lipid peroxidation of normal and sickled erythrocytes. Br J Haematol. 1980;44:87–92.[Medline]
24. Lachant NA, Tanaka KR. Antioxidants in sickle cell disease: the in vitro effects of ascorbic acid. Am J Med Sci. 1986;292:3–10.[Medline]
25. MacDonald VW, Charache S. Drug-induced oxidation and precipitation of hemoglobins A, S, and C. Biochim Biophys Acta. 1982;701:39–44.[Medline]
26. Borges A, Desforges JF. Studies of Heinz body formation. Acta Haematol. 1967;37:1–10.[Medline]
27. Bigin E, Abrams M, Earon Y. Effect of garlic extract on red blood cells. J Food Prot. 1984;47:100–1.
28. Imada O. (1990) Toxicity aspects of garlic. 1st World Congress on the Health Significance of Garlic and Garlic Constituents. Washington, D.C., 47 (abstr).
29. Kanezawa A, Nakagawa S, Sumiyoshi H, Masamoto K, Harada H, Nakagami S, Date S, Yokota A, Nishikawa M, Fuwa T. General toxicity tests of garlic extract preparation (Kyoleopin) containing vitamins. Oyo Yakuri. 1984;27:909–29.
30. Kazutani S. On effects of garlic (Allium Scorodoprasum L.) on anemia. Clin Pathol Hematol. 1934;3:1175–223.
31. Miyamoto T. Effects of garlic on hemograms. J Machurian Med. 1935;22:379–86.
32. Nakagawa S, Masamoto K, Sumiyoshi H, Kunihiro K, Fuwa T. Effect of raw and extracted-aged garlic juice on growth of young rats and their organs after peroral administration (author's transl). J Toxicol Sci. 1980;5:91–112.[Medline]
33. Freeman F, Kodera Y. Garlic chemistry: stability of S-(2-propenyl)-2-propene-1-sulfinothioate (allicin) in blood, solvents, and simulated physiological fluids. J Agric Food Chem. 1995;43:2332–8.
34. Ohnishi ST, Ohnishi T. In vitro effects of aged garlic extract and other nutritional supplements on sickle erythrocytes. J Nutr. 2001;131:1085S–92S.
35. Ohnishi ST, Ohnishi T, Ogunmola GB. Sickle cell anemia: a potential nutritional approach for a molecular disease. Nutrition. 2000;16:330–8.[Medline]
36. Imai J, Ide N, Nagae S, Moriguchi T, Matsuura H, Itakura Y. Antioxidant and radical scavenging effects of aged garlic extract and its constituents. Planta Med. 1994;60:417–20.[Medline]
37. Moriguchi T, Takasugi N, Itakura Y. The effects of aged garlic extract on lipid peroxidation and the deformability of erythrocytes. J Nutr. 2001;131:1016S–9S.
38. Glader BE, Nathan DG. Cation permeability alterations during sickling: relationship to cation composition and cellular hydration of irreversibly sickled cells. Blood. 1978;51:983–9.
39. Ohnishi ST, Katagi H, Katagi C. Inhibition of the in vitro formation of dense cells and of irreversibly sickled cells by charybdotoxin, a specific inhibitor of calcium-activated potassium efflux. Biochim Biophys Acta. 1989;1010:199–203.[Medline]
40. Ballas SK, Smith ED. Red blood cell changes during the evolution of the sickle cell painful crisis. Blood. 1992;79:2154–63.
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