Antioxidant effects of dexmedetomidine against hydrogen peroxide-induced DNA damage in vitro by alkaline Comet assay
Antioxidant effects of dexmedetomidine against hydrogen peroxide-induced DNA damage in vitro by alkaline Comet assay
Background/aim: Dexmedetomidine (DEX) is an alpha-2 adrenergic agonist that is commonly used as a sedative and anesthetic. Theprotective effects of DEX against oxidative damage under both in vitro and in vivo conditions have been demonstrated. It was aimed to evaluate and compare the protective effects of DEX and vitamin C (Vit C) on DNA against $H_2O_2$-induced DNA damage in human lymphocyte cell cultures in vitro by alkaline Comet assay. Materials and methods: Lymphocyte cell cultures were divided into 5 groups, as the negative control, solvent control, positive control, hydrogen peroxide ($H_2O_2$; 150 µM) + DEX (1 µM; 2.5 µM; 5 µM), and $H_2O_2$ (150 µM) + Vit C (1 µM; 2.5 µM; 5 µM), and incubated at37 °C for 1 h. Cell viability was measured using the Trypan blue test. DNA damage was measured using the Alkali Comet Technique and the % percent tail intensity was evaluated. Statistical analysis was performed using 1-way ANOVA and the Tukey multiple comparisontest. Results: It was observed that $H_2O_2$ significantly induced DNA damage in the lymphocytes and this damage was decreased significantly with Vit C and DEX. It was observed that Vit C at doses of 1 µM and 2.5 µM had a significantly stronger antioxidant effect, but there was no significant difference between the antioxidant effects of Vit C and DEX with a dose of 5 µM. The dose of 5 µM DEX was found to be the most effective in reducing oxidative DNA damage. Conclusion: There is limited data on the protective effects of DEX against oxidative DNA damage. The primary effect might be cytoprotection. The results herein showed that DEX was protective against $H_2O_2$-induced in vitro oxidative DNA damage in lymphocyte cell cultures in a dose-dependent manner. DEX might have a potential therapeutic value in the prevention of oxidative DNA damagein patients.
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- 1. Hall JE, Uhrich TD, Barney JA, Arain SR, Ebert TJ. Sedative, amnestic, and analgesic properties of small-dose dexmedetomidine infusions. Anesthesia and Analgesia 2000; 90 (3): 699-705. doi: 10.1097/00000539-200003000-00035
- 2. Law SW, Alexander E. Dexmedetomidine use in critical care. AACN Advanced Critical Care 2008; 19 (2): 113-120. doi: 10.1097/01.AACN.0000318109.06073.35
- 3. Ibacache M, Sanchez G, Pedrozo Z, Galvez F, Humeres C et al. Dexmedetomidine preconditioning activates pro-survival kinases and attenuates regional ischemia/reperfusion injury in rat heart. Biochimica et Biophysica Acta 2012; 1822 (4): 537- 545. doi: 10.1016/j.bbadis.2011.12.013
- 4. Gao JM, Meng XW, Zhang J, Chen WR, Xia F et al. Dexmedetomidine protects cardiomyocytes against hypoxia/ reoxygenation injury by supressing TLR4-MyD88-NF-κB signaling. BioMed Research International 2017:1674613. doi: 10.1155/2017/1674613
- 5. Gu J, Chen J, Xia P, Tao G, Zhao H et al. Dexmedetomidine attenuates remote lung injury induced by renal ischemiareperfusion in mice. Acta Anaesthesiologica Scandinavica 2011; 55 (10): 1272-1278. doi: 10.1111/j.1399-6576.2011.02526.x
- 6. Shen J, Fu G, Jiang L, Xu J, Li L et al. Effect of dexmedetomidine pretreatment on lung injury following intestinal ischemiareperfusion. Experimental and Therapeutic Medicine 2013; 6 (6): 1359-1364. doi: 10.3892/etm.2013.1317
- 7. Cui J, Zhao H, Yi B, Zeng J, Lu K et al. Dexmedetomidine attenuates bilirubin-induced lung alveolar epithelial cell death in vitro and in vivo. Critical Care Medicine 2015; 43 (9): 356- 368. doi: 10.1097/CCM.0000000000001035
- 8. Si Y, Bao H, Han L, Shi H, Zhang Y et al. Dexmedetomidine protects against renal ischemia and reperfusion injury by inhibiting the JAK/STAT signaling activation. Journal of Translational Medicine 2013; 11: 141. doi: 10.1186/1479-5876- 11-141
- 9. Freeman KA, Fullerton DA, Foley LS, Bell MT, Cleveland JC Jr et al. Spinal cord protection via alpha-2 agonistmediated increase in glial cell-line-derived neurotrophic factor. The Journal of Thoracic and Cardiovascular Surgery 2015; 149 (2): 578-584. doi: 10.1016/j.jtcvs.2014.10.037
- 10. Zhang Y, Kimelberg HK. Neuroprotection by alpha 2-adrenergic agonists in cerebral ischemia. Current Neuropharmacology 2005; 3 (4): 317-323. doi: 10.2174/157015905774322534
- 11. Okada H, Kurita T, Mochizuki T, Morita K, Sato S et al. The cardioprotective effect of dexmedetomidine on global ischaemia in isolated rat hearts. Resuscitation 2007; 74 (3): 538-545. doi: 10.1016/j.resuscitation.2007.01.032
- 12. Zhang XY, Liu ZM, Wen SH, Li YS, Li Y et al. Dexmedetomidine administration before, but not after, ischemia attenuates intestinal injury induced by intestinal ischemia-reperfusion in rats. Anesthesiology 2012; 116 (5): 1035-1046. doi: 10.1097/ ALN.0b013e3182503964
- 13. Gu J, Sun P, Zhao H, Watts HR, Sanders RD et al. Dexmedetomidine provides renoprotection against ischemia reperfusion injury in mice. Critical Care 2011; 15 (3): R153. doi: 10.1186/cc10283
- 14. Chang Y, Huang X, Liu Z, Han g, Huang L et al. Dexmedetomidine inhibits the secretion of high mobility group box 1 from lipopolysaccharide-activated macrophages in vitro. Journal of Surgical Research 2013; 181 (2): 308-314. doi: 10.1016/j.jss.2012.07.017
- 15. Schoeler M, Loetscher PD, Rossaint R, Fahlenkamp AV, Eberhardt G et al. Dexmedetomidine is neuroprotective in an in vitro model for traumatic brain injury. BMC Neurology 2012; 12:20. doi: 10.1186/1471-2377-12-20
- 16. Devasagayam TP, Tilak JC, Boloor KK, Sane KS, Ghaskadbi SS et al. Free radicals and antioxidants in human health: current status and future prospects. The Journal of the Association of Physicians of India 2004; 52: 794-804.
- 17. Song Q, Gou WL, Zhang R. FAM3A protects HT22 cells against Hydrogen peroxide-Induced Oxidative Stress through activation of PI3K/Akt but not MEK/ERK Pathway. Cellular Physiology and Biochemistry 2015; 37 (4): 1431-1441. doi: 10.1159/000438512
- 18. Griffin S. DNA damage, DNA repair and disease. Current Biology 1996; 6 (5): 497-499. doi: 10.1016/s0960- 9822(02)00525-0
- 19. Vani K, Kurakula M, Syed R, Alharbi K. Clinical relevance of vitamin C among lead-exposed infertile men. Genetic Testing and Molecular Biomarkers 2012; 16 (9): 1001-1006. doi: 10.1089/gtmb.2012.0027
- 20. Collins AR. The Comet assay for DNA damage and repair: principles, applications, and limitations. Molecular Biotechnology 2004; 26 (3): 249-261. doi: 10.1385/MB:26:3:249
- 21. Panda SK, Ravindran B. Isolation of Human PBMCs. Bioprotocol 2013; 3 (3): e323. doi: 10.21769/BioProtoc.323
- 22. Yip DK, Auersperg N. The dye-exclusion test for cell viability: persistence of differential staining following fixation. In Vitro 1972; 7 (5): 323-329. doi: 10.1007/bf02661722
- 23. Singh NP, McCoy MT, Tice RR, Schneider EL. A simple technique for quantitation of low levels of DNA damage in individual cells. Experimental Cell Research 1988; 175 (5): 184-191. doi: 10.1016/0014-4827(88)90265-0
- 24. Huet O, Dupic L, Harrois A, Duranteau J. Oxidative stress and endothelial dysfunction during sepsis. Frontiers in Bioscience 2011; 16: 1986-1995. doi: 10.2741/3835
- 25. Marnett LJ. Oxyradicals and DNA damage. Carcinogenesis 2000; 21 (3): 361-370. doi: 10.1093/carcin/21.3.361
- 26. Zhang F, Ding T, Yu L, Zhong Y, Dai H et al. Dexmedetomidine protects against oxygen-glucose deprivation-induced injury through the imidazoline receptor-PI3K/AKT pathway in rat C6 glioma cells. Journal of Pharmacy and Pharmacolology 2012; 64 (1): 120-127. doi: 10.1111/j.2042-7158.2011.01382.x
- 27. Yoon JY, Park JH, Kim EJ, Park BS, Yoon JU et al. Dexmedetomidine attenuates $H_2O_2$-induced cell death in human osteoblasts. Journal of Dental Anesthesia and Pain Medicine 2016; 16 (4): 295-302. doi: 10.17245/ jdapm.2016.16.4.295
- 28. Cai Y, Xu H, Yan J, Zhang L, Lu Y. Molecular targets and mechanism of action of dexmedetomidine in treatment of ischemia/reperfusion injury. Molecular Medicine Reports 2014; 9 (5): 1542-1550. doi: 10.3892/mmr.2014.2034
- 29. Nakamura H, Nakamura K, Yodoi J. Redox regulation of cellular activation. Annual Review of Immunology 1997; 15: 351-369. doi: 10.1146/annurev.immunol.15.1.351