Use and Evaluation of Newly Synthesized Fluorescence Probes to Detect Generated OH• Radicals in Fibroblast Cells

Overview

Journal J Fluoresc

Specialties Biophysics
Chemistry

Date 2016 Mar 18

PMID 26983614

Citations 2

Authors

Reza Salimi

Nilgun Yener

Roghaiyeh Safari

Affiliations

Soon will be listed here.

Abstract

Reactive oxygen species (ROS) are pro-oxidant molecules synthesized in body with various functions and are essential for life. Increasing in reactive oxygen species or decreasing in antioxidants level cause oxidative stress which is very harmful. OH• radical is one of ROS's, with tendency to bind to lipids, DNA and proteins which cause irreversible damage in cells. The most devastating consequences related to excess OH• radicals occur via direct binding to nucleic acids and proteins. Quantification of this high reactive radical with short life time is difficult. Electron Spin Resonance, Fluorescence, and Luminescence Spectroscopy are commonly used to determine the level of ROS. Fluorescence Probes have higher specificity and sensitivity with their excellent sensors to detect ROS's compare to the other methods. Also, there are different probes specifically designed for each radical. The purpose of this study was to identify the probe better suiting for detection of OH• radical levels. The two most recommended fluorescence probes, 2-[6-(4 V-Hydroxy) phenoxy-3H-xanthen-3-on-9-yl]benzoic acid (HPF) and coumarin-3-carboxylic acid (3-CCA) to determine OH• radical levels were compared. Following the formation of OH• radical with Fenton reaction, HPF and 3-CCA probes were added to cells and spectrofluorometric measurements were performed in their respective wavelengths. The mean amplitude of fluorescence for HPF was 32.72 ± 2.37 F.I (n = 40) and for 3-CCA was 52.11 ± 0.5 F.I (n = 40). This difference was statistically significant. 3-CCA also demonstrated more stable measurements at different days compered to HPF.

Citing Articles

Preparation of Electrospun Active Molecules Membrane Application to Atmospheric Free Radicals.

Yang Y, Wang G, Li X, Iradukunda Y, Liu F, Li Z Membranes (Basel). 2022; 12(5).

PMID: 35629806 PMC: 9143268. DOI: 10.3390/membranes12050480.

Changes in the Spectral Features of Zinc Phthalocyanine Induced by Nitrogen Dioxide Gas in Solution and in Solid Polymer Nanofiber Media.

Zugle R, Tetteh S J Fluoresc. 2016; 27(2):739-743.

PMID: 27987103 DOI: 10.1007/s10895-016-2006-x.

References

Setsukinai K, Urano Y, Kakinuma K, Majima H, Nagano T . Development of novel fluorescence probes that can reliably detect reactive oxygen species and distinguish specific species. J Biol Chem. 2002; 278(5):3170-5. DOI: 10.1074/jbc.M209264200. View

Ou B, Hampsch-Woodill M, Flanagan J, Deemer E, Prior R, Huang D . Novel fluorometric assay for hydroxyl radical prevention capacity using fluorescein as the probe. J Agric Food Chem. 2002; 50(10):2772-7. DOI: 10.1021/jf011480w. View

Indo H, Davidson M, Yen H, Suenaga S, Tomita K, Nishii T . Evidence of ROS generation by mitochondria in cells with impaired electron transport chain and mitochondrial DNA damage. Mitochondrion. 2007; 7(1-2):106-18. DOI: 10.1016/j.mito.2006.11.026. View

Halliwell B, Whiteman M . Measuring reactive species and oxidative damage in vivo and in cell culture: how should you do it and what do the results mean?. Br J Pharmacol. 2004; 142(2):231-55. PMC: 1574951. DOI: 10.1038/sj.bjp.0705776. View

Lemire J, Harrison J, Turner R . Antimicrobial activity of metals: mechanisms, molecular targets and applications. Nat Rev Microbiol. 2013; 11(6):371-84. DOI: 10.1038/nrmicro3028. View

Tai C, Gu X, Zou H, Guo Q . A new simple and sensitive fluorometric method for the determination of hydroxyl radical and its application. Talanta. 2008; 58(4):661-7. DOI: 10.1016/s0039-9140(02)00370-3. View

Bromme H, Zuhlke L, Silber R, Simm A . DCFH2 interactions with hydroxyl radicals and other oxidants--influence of organic solvents. Exp Gerontol. 2008; 43(7):638-644. DOI: 10.1016/j.exger.2008.01.010. View

Moore J, Yin J, Yu L . Novel fluorometric assay for hydroxyl radical scavenging capacity (HOSC) estimation. J Agric Food Chem. 2006; 54(3):617-26. DOI: 10.1021/jf052555p. View

Ziech D, Franco R, Georgakilas A, Georgakila S, Malamou-Mitsi V, Schoneveld O . The role of reactive oxygen species and oxidative stress in environmental carcinogenesis and biomarker development. Chem Biol Interact. 2010; 188(2):334-9. DOI: 10.1016/j.cbi.2010.07.010. View

10.

Yang X, Guo X . Investigation of the anthracene-nitroxide hybrid molecule as a probe for hydroxyl radicals. Analyst. 2001; 126(10):1800-4. DOI: 10.1039/b103208a. View

11.

Filaire E, Toumi H . Reactive oxygen species and exercise on bone metabolism: friend or enemy?. Joint Bone Spine. 2012; 79(4):341-6. DOI: 10.1016/j.jbspin.2012.03.007. View

12.

Soh N, Makihara K, Ariyoshi T, Seto D, Maki T, Nakajima H . Phospholipid-linked coumarin: a fluorescent probe for sensing hydroxyl radicals in lipid membranes. Anal Sci. 2008; 24(2):293-6. DOI: 10.2116/analsci.24.293. View

13.

Soh N . Recent advances in fluorescent probes for the detection of reactive oxygen species. Anal Bioanal Chem. 2006; 386(3):532-43. DOI: 10.1007/s00216-006-0366-9. View

14.

Manevich Y, Held K, Biaglow J . Coumarin-3-carboxylic acid as a detector for hydroxyl radicals generated chemically and by gamma radiation. Radiat Res. 1997; 148(6):580-91. View

15.

Fang Y, Yang S, Wu G . Free radicals, antioxidants, and nutrition. Nutrition. 2002; 18(10):872-9. DOI: 10.1016/s0899-9007(02)00916-4. View

16.

Gomes A, Fernandes E, Lima J . Fluorescence probes used for detection of reactive oxygen species. J Biochem Biophys Methods. 2005; 65(2-3):45-80. DOI: 10.1016/j.jbbm.2005.10.003. View

17.

Brandes R, Janiszewski M . Direct detection of reactive oxygen species ex vivo. Kidney Int. 2005; 67(5):1662-4. DOI: 10.1111/j.1523-1755.2005.00258.x. View

18.

Grankvist K, Marklund S, Sehlin J, Taljedal I . Superoxide dismutase, catalase and scavengers of hydroxyl radical protect against the toxic action of alloxan on pancreatic islet cells in vitro. Biochem J. 1979; 182(1):17-25. PMC: 1161229. DOI: 10.1042/bj1820017. View

19.

Xiao H, Parkin K . Antioxidant functions of selected allium thiosulfinates and S-alk(en)yl-L-cysteine sulfoxides. J Agric Food Chem. 2002; 50(9):2488-93. DOI: 10.1021/jf011137r. View

20.

Bartosz G . Use of spectroscopic probes for detection of reactive oxygen species. Clin Chim Acta. 2006; 368(1-2):53-76. DOI: 10.1016/j.cca.2005.12.039. View