Investigation of DNA Damage in Rats in Chronically Exposed to Sub-Hypnotic Dose Desflurane

Yıl 2015, Cilt: 12 Sayı: 1, 44 - 52, 15.04.2015

Nuray Altay Zeynep Baysal Yıldırım

Öz

Background: In this study the aim was to research the DNA damage in rats whose chronic desfluran gases
exposed.
Metods: This experimental study, after the Ethic Committe permission was made in Harran University
Medical Faculty Farmacology Department. For the study 200-230 gr 18 female albino wistar rats were used.
After and before the study care and the feeding of the rats were made in the Harran University experimental
animal laboratory. Rats divided in two groups (Group I: Desfluran, Grup II: Control ). For the anaesthesia
pump system we used special planned plastic plate. Datex Ohmeda Tec 6 plus vaporizator was used to Group
I rats to whom hypnotic doses desflurane was given ( % 0,5-1 ). İn the second group same mechanism was
used but the desflurane vaporizator was closed. Each group %50 O2 and desflurane gas mixture was given in
the 5lt/minute speed. In the environment the desflurane and the O2 concentration were measured continously
by the anaesthesia gases monitor ( criticare ). The condition of exposing to the gases, again 1 hour in 5 days.
Outside of experiment, rats waited in room condition and in their cages. At the end of fifth day, 50mg/kg with
penthotal sodium, general anaesthesia was made and their extremities were fixed in the supine position fixing
to the table. Blood samples were taken from intracardiac undertaking. The blood samples were taken
heparinized tubes and put in cold condition in the special plate. Then sent to the Harran University Medical
Faculty Biochemical Department. The experimental rats sacrificed and then burried with lime. DNAdamage
was measured in taken bloods mononuclear leukocytes with Comet Assay method. İn addition, in same blood
samples were measured antioxidant capacity and oxidative stress.
Results: During the study, general anaesthesia conditions in any rat were not seen and rats were continued
their normal activity. DNAdamage, antioxidant capacity and oxidative stress were similar in both groups.
Conclusions: It was concluded that not occur in a significant DNA damage, compared to the control
group in rats exposed chronically desflurane.

Anahtar Kelimeler

Desflurane, chronic exposure, DNAdamage

Sıçanlarda, Subhipnotik Doz Kronik Desfluran Maruziyetinde, DNA Hasarının Araştırılması

Öz

Amaç: Bu çalışmada, subhipnotik dozda, kronik desfluran gazına maruz kalan sıçanlardaki DNA hasarının
araştırılması amaçlandı.
Materyal ve metod: Deneysel çalışma, Harran Üniversitesi Tıp Fakültesi Etik Kurulu izni sonrasında aynı
fakültenin Farmakoloji Laboratuarında yapıldı. Çalışma için 200-230gr ağırlığında 18 adet dişi Wistar türü
albino sıçan kullanıldı. Sıçanların çalışma öncesi ve sonrasındaki beslenme ve bakımları, Harran
Üniversitesi Deney Hayvanları Laboratuarında yapıldı. Sıçanlar randomize 2 gruba ayrıldı (Grup
I=Desfluran, Grup II= Kontrol). Anestezik gaz verme sistemi için, özel olarak tasarlanmış plastik kaplar
kullanıldı. Grup I' de sıçanlara, Datex-Ohmeda Tec6 Plus vaporizatörü ile hipnoz oluşturmayacak dozlarda
(%0.5-1) desfluran verildi. Grup II'de de aynı düzenek kullanıldı, ancak desfluran vaporizatörü kapalı
tutuldu. Her iki grupta deney düzeneğine %50 oksijen-hava karışımı 5 litre /dakika taze gaz akışı hızında
verildi. Ortamdaki desfluran ve oksijen konsantrasyonları bir anestezik gaz monitörü (Criticare ®) ile sürekli
olarak ölçüldü. Çalışma gazlarına maruz kalma durumu 5 gün boyunca günde 1 saat olacak şekilde
tekrarlandı. Deney dışındaki saatlerde sıçanlar kendi kafeslerinde oda şartlarında bakıldılar. Beşinci günün
sonunda 50 mg/kg intraperitoneal pentothal ile anestezisi sağlanan sıçanlar, supin pozisyonunda
ekstremitelerden masaya tespit edildi. Sıçanlardan intrakardiak girişimle kan örnekleri alındı. Alınan kanlar
heparinli tüplere aktarılarak özel bir kap içerisinde, soğuk ortamda, en kısa sürede Harran Üniversitesi Tıp
Fakültesi Biyokimya Anabilim Dalı Laboratuarına ulaştırıldı. Deney hayvanları kan alındıktan sonra
sakrifiye edilip, üzerlerine kireç kaymağı dökülüp toprağa gömüldüler. Alınan kanlardaki mononükleer
lökositlerinden Comet Assey yöntemi ile DNAhasarları ölçüldü. Ayrıca aynı kan örneklerinden, antioksidan
kapasite ve oksidatif stress indeks değerleri analiz edildi.
Bulgular: Deneysel çalışma sırasında hiçbir sıçanda genel anestezi durumu oluşmazken, tüm sıçanlar 5 gün
boyunca normal aktivitelerini sürdürdüler. Kan örneklerinden çalışılan DNAhasarı, antioksidan kapasite ve
oksidatif stres indeks değerleri, her iki grupta benzer bulundu.

Sonuç: Kronik olarak desflurana maruz kalan sıçanlarda, kontrol grubuna göre, belirgin bir DNA hasarının
oluşmadığı sonucuna varıldı.

Anahtar Kelimeler

Desfluran, kronik maruziyet, DNAhasarı

Kaynakça

• 1) Kayhan Z. Klinik Anestezi. Logos Yayıncılık Tic. Aş. Ankara, 1997: 56. 2) Ronald D. Miller:Anesthesia, Churchill Livingstone, Philadelphia, 2000: 166. 3) Hoerauf K.H, Schrögendorfer K.F, Wiesner G, Gruber M, Spacek A, Kress H.G and Rüdiger H.W. Sister chromatid exchange in human lymphocytes exposed to isoflurane and nitrous oxide in vitro. British Journal of Anesthesia 1999; 82(2):268-70. 4) Karabıyık L, Şardaş S, Polat U, Kocabaş N.A, Karakaya A.E. Comparison of genotoxicity of sevoflurane and isoflurane in human lymphocytes studied in vivo using the comet assay. Mutation Research 2001;497(1-2):99-107.De Hert SG 5) Van der Linden PJ, ten Broecke PW, Vermeylen KT, Rodrigus IE, Stockman BA. Effects of desflurane and sevoflurane on length-dependent regulation of myocardial function in coronary surgery patients. Anesthesiology 2001;95(2):357-63. 6) Kharasch ED, Thummel KE. Identification of cytochrome P450 2E1 as the predominant enzyme catalyzing human liver microsomal defluorination of sevoflurane, isoflurane, and methoxyflurane. Anesthesiology 1993;79(4):795-807. 7) Özatamer O, Alkış N, Batislam Y. Fast-Tracking. Anestezide Güncel Konular, Ankara, 2002:417. 8) Patel SS, Goa KL. Desflurane. A review of its pharmacodynamic and pharmacokinetic properties and its efficacy in general anaesthesia. Drug 1995;50(4):742- 67. 9) Halliwell B. Reactive oxygen species in living systems: source, biochemistry, and role in human disease. Am J Med 1991;91(3):14-22. 10) Kayhan Z. Klinik Anestezi, Logos Yayıncılık Tic. Aş. Ankara, 1997, 270-91. 11) Muggli D. Physiological requrements of vitamen E as a function of the amount and type of polyunsatured fatty acid. World Rev Nutr Diet 1994;75:166-8.Murphy PG 12) Bennett JR, Myers DS, Davies MJ, Jones JG. The effect of propofol anaesthesia on free radical-induced lipid peroxidation in rat liver microsomes. Eur J Anaesthesiol 1993;10(4):261-66.Rosenberg PH 13) KallioH. Operating-theatre gas pollution and chromosomes. Lancet 1977;2(8035):452-3. 14) Ronald D. Miller:Anesthesia. Churchill Livingstone. Philadelphia, 2000, 167.Hoerauf KH 15) Wiesner G, Schroegendorfer KF, Jobst BP, Spacek A, Harth M, Sator-Katzenschlager S, Rüdiger HW. Waste anaesthetic gases induce sister chromatid exchanges in lymphocytes of operating room personel. British.Journal of Anaesthasia 1999;82(5):764-66. 16) Karpinski T.M, Kostrzewska-Poczekaj M, Stachecki I, Mikstacki A, Szyfter K. Genotoxicity of the volatile anaesthetic desflurane in human lymphocytes in vitro established by comet assay . J.Appl Genet 2005;46(3):319-24. 17) Szyfter K, Szule R, Mikstacki A, Stachecki I, Rydzanicz M, Jaloszynski P. Genotoxicity of inhalation anaesthetics:DNAlesions generated by sevoflurane in vitro and in vivo. J.Appl.Genet 2004;45(3):369-74.Bilban M 18) Jakopin CB, Ogrinc D. Cytogenetic tests performed on operating room personel(the use of anaesthetic gases). Int Arch Occup Environ Health 2005;78(1):60-4. 19) Natarajan D, Santhiya ST. Cytogenetic damage in operation theatre personel. Anaesthesia 1990;45(7):574-77. 20) Husum B, Nieburh E, Wulf HC, Norgaard I. Sister chromatid exchanges and structural chromosome aberrations in lymphocytes in operating room personel. Acta Anaesthesiology Scand 1983;27(3):262-65. 21) Hoerauf K, Lierz M, Wiesner G, Schroegendorfer K, Lierz P, Spacek A, Brunnberg L, Nusse M. Genetic damage in operating room personel exposed to ısoflurane and nitrous oxide. Occup Environ Med 1999;56(7):433-37. Sardaş S 22) Aygün N, Gamli M, Unal Y, Unal N, Berk N, Karakaya AE. Use of alkaline comet assay to detect DNA damages in lymphocytes of operating room personel occupationally exposed to anaesthetic gases. Mutat Res 1998;418(2-3):93- 100. Bruce DL 23) Eide KA, Linde HW, Eckenhoff JE. Causes of death among anesthesiologists: a 20-year survey. Anesthesiology 1968;29(3):565-9. 24) Rozdaj R, Kasuba V, Jazbec A. Preliminary study of cytogenetic damage in personel exposed to anaesthetic gases. Mutagenesis 2001;16(2):139-43. Reitz M 25) Antonini-Rumpf E, Lanz E. DNA single-strand breaks in peripheral human lymphocytes after anaesthesia with isoflurane-nitrous oxide-oxygen. Arzneimittelforschung 1993;43(12):1258-61. Jaloszyński P 26) Kujawski M, Wasowicz M, Szulc R, Szyfter K. Genotoxicity of inhalation anaesthetics halotane and isoflurane in human lymphocytes studied in vitro using the comet assay. Mutat Res 1999;439(2):199-206. Coate WB 27)Kapp RW Jr, Lewis TR. Chronic exposure to low concentrations of halotan-nitrous oxide;reproductive and cytogenetic effect in the rat. Anaesthesiology 1979; 50(4):310-8. Natarajan D 28) Santhiya ST. Cytogenetic damage in operation theatre personel. Anaesthesia 1990;45(7):574-7.Chang WP 29)Lee S, Tu J, Hseu S. Increased micronucleus formation in nurses with occupational nitrous oxide exposure in operating theaters. Environ Mol Mutagen 1996;27(2):93-7. Bigatti P 30)Lamberti L, Ardito G, Armellino F, Malanetto C. Chromosome aberrations and sister chromatid exchanges in occupationally exposed workers. Med Lav 1985;76(4):334-9. Murphy PG 31)Myers DS, Davies MJ, Webster NR, Jones JG. The a n t i o x i d a n t p o t e n t i a l o f p r o p o f o l ( 2 , 6 - diisopropylphenol). Br J Anaesth 1992; 68(6):613-18. 32)Murrell GA, Francis MJ, Bromley L. Modulation of fibroblast proliferation by oxygen free radicals. Biochem J 1990; 265(3):659-65. Musacchio E 33)Rizzoli V, Bianchi M, Bindoli A, Galzigna L. Antioxidant action of propofol on liver microsomes, mitochondria and brain synaptosomes in the rat. Pharmacol Toxicol 1991; 69(1):75-77. 34)Aebi H. Catalase in vitro. Methods Enzymol 1984;105:121-26. Ansley DM 35)Sun J, Visser WA, Dolman J, Godin DV, Garnett ME, Qayumi AK. High dose propofol enhances red cell antioxidant capacity during CPB in humans. Can J Anaesth 1999;46(7):641-48. Johnson ME 36)Sill JC, Uhl CB, Halsey TJ, Gores GJ. Effect of volatile anesthetics on hydrogen peroxide-induced injury in aortic and pulmonary arterial endothelial cells. Anesthesiology 1996;84(1):103-16. Allaouchiche B 37)Debon R, Goudable J, Chassard D, Duflo F. Oxidative stress status during exposure to propofol, sevoflurane and desflurane. Anesth Analg 2001;93(4):981-85.Koksal GM 38)Sayilgan C, Aydin S, Uzun H, Oz H. The effects of sevoflurane and desflurane on lipid peroxidation during laparoscopic cholecystectomy. Eur J Anaesthesiol 2004;21(3):217-20. Dikmen B 39)Unal Y, Pampal HK, Nurlu N, Kurtipek O, Canbolat O, Ozoğul C, Kavutcu M. Effects of repeated desflurane and sevoflurane anaesthesia on enzymatic free radikal scavanger system. Mol Cell Biochem 2007;294(1-2):31-37.