İ. Özkan ÖNAL, İ. Aydın ERDEN, Seda Banu AKINCI, Gönül ERDEN, İsmail KARABULUT, N. Dilara ZEYBEK, Ersin FADILLIOĞLU, Sevda MÜFTÜOĞLU, Fatma SARICAOĞLU, Z. Dicle BALKANCIOĞLU, K.Hüsnü Can BAŞER, Ülkü AYPAR

Sıçamlarda karvakrol'un oleik asitle oluşturulan akut akciğeri hasarına etkisi

Amaç: Bu çalışma deneysel olarak oleik asitle oluşturulan akciğer hasarında oksidan, anti-oksidan sistemlerindeki değişiklikler ile antioksidan özelliği bilinen ve daha önce akciğer, meme, karaciğer, testis tümörlerinde iyileştirici etkisi gösterilmiş olan karvakrolun (kekik suyu) olası iyileştirici ve koruyucu etkisinin araştırılması amacı ile yapılmıştır. Gereç ve Yöntem: Çalışmaya 28 adet Wistar cinsi erkek rat (250-350 gr) alındı. Ratlar 4 gruba ayrıldı. Grup I: Oleik asit grubu (n:7, 50 pil oleik asit kuyruk veni kullanılarak intravenöz (iv), 30 dakika sonra 73 mg kg'1 karvakrol dozuna eşdeğer hacimde SF intraperitoneal (ip) uygulandı). Grup II: Oleik asit+Karvakrol grubu (n:7, 50 fil oleik asit iv ve 30 dakika sonra 73 mg kg'1 karvakrol ip uygulandı). Grup III: Kontrol grubu (n: 7,50 pil SF iv, 30 dakika sonra 73 mg kg'1 karvakrol dozuna eşdeğer hacimde SF ip uygulandı). Grup IV: Karvakrol grubu (50 pil SF iv, 30 dakika sonra 73 mg kg1 dozunda karvakrol ip uygulandı). Tüm ratlar enjeksiyonlardan sonra 4 saat oksijen odasında tutuldu ve sakrifiye edildi. İncelediği sıçanların hangi gruba ait olduğunu bilmeyen iki histolog tarafından diseke edilen akciğerler akut akciğer hasarı açısından değerlendirildi. Işık mikroskobisinde interstisyel ödem, alveolar hemoraji, total akciğer hasarı, intraalveolar nötrofiller, makrofajlar, pnömositler; elektron mikroskobisinde kan-gaz bariyerinin bütünlüğü, Tip 1 ve 2 pnömositlerin morfolojisi değerlendirildi. Ayrıca akciğer dokusunda oksidan, anti-oksidan parametreler olarak kabul edilen protein karbonil, malondialdehid doku (MDA doku), katalaz (CAT), glutatyon peroksidaz (GSH-Px), süperoksit dismutaz (SOD) bakıldı. Serumda pro-inflamatuar ve anti-inflamatuar sitokinlerden interlökin-6 (IL-6), interlökin-10 (IL10), tümör nekroz naktör alfa (TNF-alfa), granülösit kolonize stimüle edici faktör (G-CSF) düzeylerine bakıldı. Bulgular: Oleik asit, kontrol ve karvakrol grupları ile karşılaştırıldığında önemli ölçüde akut akciğer hasarı oluşturdu, proteinkarbonil ve MDA doku düzeyleri daha yüksekti. Oleik asit ve Oleik asit+karvakrol grupları arasında total akciğer hasarı, protein karbonil, CAT, GSH-Px, SOD düzeyleri açısından anlamlı fark saptanmadı. Oleik asit+ karvakrol, karvakrol ve kontrol grubu doku MDA düzeyi yalnız oleik asit verilen gruba göre anlamlı olarak daha düşüktü. Pro-inflamatuar ve anti-inflamatuar sitokinlerden IL-6, IL-10, TNF-a, G-CSF düzeyleri açısından ise gruplar arasında anlamlı fark bulunmadı. Sonuç: Akut akciğer hasarında karvakrolun oksidan, anti-oksidan sistemlerindeki dengeyi anti-oksidanlar lehine arttırması nedeniyle karvakrol alımının arttırılması akciğer hasarını önlemede ve oluşmuş akciğer hasarını iyileştirmede faydalı olabilir.

The effects of carvacrol in oleic acid induced acute lung injury in rats

Objective: This study was conducted to compare the effects of oleic acid on the antioxidant systems of lung muscle tissue and carvacrol which has been proven to have healing effects on tumors of the lung, liver, breast, and testis and is known to have antioxidant effects, with respect to their healing and protective properties. Methods: 28 male rats of the Wistar type (250-350 grams) were included in the study. They were divided to four groups. Group I: Group Oleic acid (n: 7, 50 jil of oleic acid were administered intravenously (iv) and 30 minutes later 73 mg kg' of saline solution was given intraperitoneally (ip) as the same volume as carvacrol). Group II: Group Oleic acid+Carvacrol (n: 7, 50 pil of oleic acid was administered iv and 30 minutes later 73 mg kg1 of carvacrol was administered ip). Group III: Group Control (n: 7,50 yıl of saline solution (the same dose as the oleic acid group) was administered and 30 minutes later 73 mg kg'1 of saline solution was administered ip). Group IV: Group Carvacrol: (n: 7, 50 pil of iv saline solution was administered and 30 minutes later 73 mg kg' carvacrol was administered ip). After being held in the oxygen room for 4 hours, blood and tissue samples were collected from the rats. Two histologists who were unaware of the group assignments assessed the acute lung injury via light microscopy; interstitial oedema, alveolar haemorrhage, intraalveolar neutrophils, intraalveolar macrophages and intraalveolar pneumocytes and electron microscopy; the integrity of the blood-gas barrier, the morphology of the type 1 and type 2 pneumocytes, and the presence of intraalveolar macrophages and granular material. The lung tissue was examined histopathologically and levels of protein carbonyl, malondialdehid tissue, catalase (CAT), glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) which are known as oxidant, anti-oxidant parameters were measured. Levels of antiinflammatory cytokines such as interleukin (IL)-6, IL-10, Tumor necrosis factor (TNF)-a and granulocyte colony stimulating factor (G-CSF) were measured in the blood samples. Results: Oleic acid caused significant total lung injury compared to the control group and to the vehicle group and levels of protein carbonyl and malondialdehid were higher than the other groups. Total lung tissue damage score, protein carbonyl, CAT, GSH-Px, SOD, levels were found to be similar in oleic acid and oleic acid+carvacrol groups. Levels ofMDA tissue were lower in carvacrol and oleic acid+carvacrol groups than oleic acid group. Levels of proinflammatory and inflammatory cytokines IL-6, IL-10, TNF-a and G-CSF were found to be similar in both groups. Conclusion: As a result, it has been shown that because of the antidxidant effects of carvacrol in lung tissue, the increased use of carvacrol may be helpful in preventing and reversing lung tissue damage.

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1.Repine JE. Scientific perspectives on adult respiratory distress syndrome. Lancet 1992;339(8791):466-469.
2.Artigas A, Bernard GR, Carlet J et al. The American-European Consensus Conference on ARDS, part 2: Ventilatory, pharmacologic, supportive therapy, study design strategies, and issues related to recovery and remodeling. Acute respiratory distress syndrome. Am J Respir Crit Care Med 1998;157(4 Pt 1):1332-1347.
3.Bernard GR, Artigas A, Brigham KL et al. The American-European Consensus Conference on ARDS. Definitions, mechanisms, relevant outcomes, and clinical trial coordination. Am J Respir Crit Care Med 1994; 149(3 Pt l):818-824.
4.Ashbaugh DG, Bigelow DB, Petty TL, Levine BE. Acute respiratory distress in adults. Lancet 1967;2(7511):319-323.
5.Lee WL, Downey GP. Neutrophil activation and acute lung injury. Curr Opin Crit Care 2001;7(l):l-7.
6.Syrbu S, Thrall RS, Smilowitz HM. Sequential appearance of inflammatory mediators in rat bronchoalveolar lavage fluid after oleic acid-induced lung injury. Exp Lung Res 1996;22(l):33-49.
7.Köksel O, Cinel I, Tamer L, et al. N-acetylcysteine inhibits peroxynitrite-mediated damage in oleic acid-induced lung injury. Pulm Pharmacol Ther 2004;17(5):263-270.
8.Rylander C, Hogman M, Perchiazzi G, Magnusson A, Hedenstierna G. Oleic acid lung injury: a morphometric analysis: using computed tomography. Acta Anaesthesiol Scand 2004;48(9): 1123-1129.
9.Schuster DP. ARDS: clinical lessons from the oleic acid model of acute lung injury. Am J Respir Crit Care Med 1994;149(1):245-260.
10.Eiermann GJ, Dickey BF, Thrall RS. Polymorphonuclear leukocyte participation in acute oleic-acid induced lung injury. Am Rev Respir Dis 1983;128(5):845-850.
11.Lang JD, Mcardle PJ, O'Reilly PJ, Matalon S. Oxidant-antioxidant balance in acute lung injury. Chest 2002;122(Suppl 6):314-320.
12.Liu H, Zhang D, Zhao B, Zhao J. Superoxide anion, the main species of ROS in the development of ARDS induced by oleic acid. Free Radic Res 2004;38(12):1281-1287.
13.Ito K, Mizutani A, Kira S, Mori M, Iwasaka H, Noguchi T. Effect of Ulinastation, a human urinary trypsin inhibitor, on the oleic acid-induced acute lung injury in rats via the inhibition of activated leukocytes. Injury 2005;36(3):387-394.
14.Xie LX, Liu YN, Zhao XW, et al. Effects of prone position ventilation on inflammatory factors in blood and bronchial alveolar lavage fluid of acute respiratory distress syndrome dogs caused by pulmonary and extrapulmonary insults. Zhonghua Yi Xue Za Zhi 2004;84(14): 1200-1204.
15.Xu GL, Yao L, Rao SY, Gong ZN, Zhang SQ, Yu SQ. Attenuation of acute lung injury in mice by oxymatrine is associated with inhibition of phosphorylated p38 mitogen-activated protein kinase. J Ethnopharmacol 2005;98(1-2):177-183.
16.Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193(l):265-275.
17.Azaz D, Demirci F, Satil F, Kürkcüoglu M, Başer KH. Antimicrobial activity of some Satureja essential oils. Z Naturforsch C 2002; 57(7-9):817-821.
18.Yanishlieva NV, Marinova EM, Gordon MH, Raneva VG. Antioxidant activity and mechanisms of action of thymol and carvacrol in two lipid system. Food Chem 1999;64(l):59-66.
19.Arunasree KM. Anti-proliferative effects of carvacrol on a human metastatic breast cancer cell line, MDA-MB 231. Phytomedicine 2010; 17(8-9):581-588.
20.Fiorentino DF, Bond MW, Mosmann TR. Two types of mouse T helper cell. IV. Th2 clones secrete a factor that inhibits cytokine production by Thl clones. J Exp Med 1989;170(6):2081-2095.
21.Fiorentino DF, Zlotnik A, Mosmann TR, Howard M, O'Garra A. IL-10 inhibits cytokine production by activated macrophages. J Immunol 1991;147(ll):3815-3822.
22.Bogdan C, Vodovotz Y, Nathan C. Macrophage deactivation by interleukin-10. J Exp Med 1991; 174(6): 1549-1555.
23.Armstrong L, Millar AB. Relative production of tumor necrosis factor alpha and -interleukin 10 in adult respiratory distress syndrome. Thorax 1997;52(5):442-446.
24.Sun Y, Oberley LW, Li Y. A simple method for clinical assay of superoxide dismutase. Clin Chem 1988;34(3):497-500.
25.Aebi H. Catalase. Bergmeyer HU (ed) Methods of enzymatic analysis Academic, New York, (1974) pp 673-677.
26.Chen HI, Hsieh NK, Kao SJ, Su CF. Protective effects of propofol on acute lung injury induced by oleic acid in conscious rats. Crit Care Med 2008;36(4): 1214-1221.
27.Kennedy MT, Higgins BD, Costello JF, Curtin WA, Laffey JG. Hypertonic saline reduces inflammation and enhances the resolution of oleic acid induced acute lung injury. BMC Pulm Med 2008;8:9.
28.Zhu GF, Sun B, Niu SF, et al. Combined surfactant therapy and inhaled nitric oxide in rabbits with oleic acid-induced acute respiratory distress syndrome. Am J Respir Crit Care Med 1998;58(2):437-443.
29.Koparal AT, Zeytinoglu M. Effects of Carvacrol on a Human Non-Small Cell Lung Cancer (NSCLC) Cell Line, A549. Cytotechnology 2003 ;43(1-3): 149-154.
30.McGuigan RM, Mullenix P, Norlund LL, Ward D, Walts M, Azarow K. Acute lung injury using oleic acid in the laboratory rat: establishment of a working model and evidence against free radicals in the acute phase. Curr Surg 2003;60(4):412-417.
31.ipek E, Zeytinoglu H, Okay S, Tuylu BA, Kurkcuoglu M, Baser HC. Genotoxicity and antigenotoxicity of Origanum oil and carvacrol evaluated by Ames Salmonella/microsomal test. Food Chem 2005;93(3):551-556.
32.Meyancı G, Ancıoglu F, Oz H, Aydemir A. Akut akciğer hasarında intratrakeal deksametazonun lipid peroksidasyonuna etkileri. Cerrahpaşa J Med 2001;32(l):20-24.
33.Meyancı KG, Sayılgan C, Find A, Uzan S, Oz H. Akut akciğer hasarının tedavisinde erken dönemde intratrakeal PG El'in lipid peroksidasyonu üzerine etkisi. GKD Anestezi Yoğun Bakım Derneği Dergisi 2004;10(3):108-110.
34.Arunasree KM. Anti-proliferative effects of carvacrol on a human metastatic breast cancer cell line, MDA-MB 231. Phytomedicine 2010;17(8-9):581-588.
35.Velmurugan B, Nagini S. Combination chemoprevention of experimental gastric carcinogenesis by s-allylcysteine and lycopene: modulatory effects on glutathione redox cycle antioxidants. J Med Food 2005;8(4):494-501.
36.Velmurugan B, Bhuvaneswari V, Burra UK, Nagini S. Prevention of N-methyl- N'-nitro-N-nitrosoguanidine and saturated sodium chloride-induced gastric carcinogenesis in Wistar rats by lycopene. Eur J Cancer Prev 2002; 11(1): 19-26.
37.Liu H, Zhang D, Zhao B, Zhao J. Superoxide anion, the main species of ROS in the development of ARDS induced by oleic acid. Free Radic Res 2004;38(12):1281-1287.
38.Leff JA, Parsons PE, Day CE, et al. Serum antioxidants as predictors of adult respiratory distress syndrome in patients with sepsis. Lancet 1993;341(8848):777-780.
39.Leff JA, Parsons PE, Day CE, et al. Increased serum catalase activity in septic patients with the adult respiratory distress syndrome. Am Rev Respir Dis 1992;146(4):985-989.
40.Brigham KL. Role of free radicals in lung injury. Chest 1986; 89(6):859-863.
41.Goodman RB, Pugin J, Lee JS, Matthay MA. Cytokine-mediated inflammation in acute lung injury. Cytokine and Growth Factor Rev 2003; 14(6):523-535.
42.Koksel O, Cinel I, Tamer L, et al. N-acetylcysteine inhibits peroxynitrite-mediated damage in oleic acid-induced lung injury. Pulm Pharmacol Ther 2004;17(5):263-270.
43.McGuigan RM, Mullenix P, Norlund LL, Ward D, Walts M, Azarow K. Acute lung injury using oleic acid in the laboratory rat: establishment of a working model and evidence against free radicals in the acute phase. Curr Surg 2003;60(4):412-417.