Sensitive fluorometric determination of glutathione using fluorescent polymer dots and the dopamine-melanin nanosystem

Microchimica Acta volume 186, Article number: 568 (2019) Cite this article

Abstract

A bioinspired fluorometric method has been developed for the detection of glutathione (GSH) in biological fluids. It is based on the use of near-infrared fluorescent semiconducting polymer dots (P-dots) and of the dopamine (DA)-melanin nanosystem. The P-dots were prepared from poly(styrene-co-maleic anhydride), the semiconducting polymer poly[(9,9′-dioctyl-2,7-divinylenefluorenylene)-alt-2-methoxy-5-(2-ethyl-hexyloxy)-1,4-phenylene] and the fluorescent dye tetraphenylporphyrin. They have excitation/emission maxima at 458/656 nm, and this enables measurement to be performed with low autofluorescence and scattering background. DA can self-polymerize on the surface of the P-dots to yield a poly-DA coating. This coating, at weak alkaline pH values, causes the quenching of the fluorescence of the P-dots. However, the polymerization of DA is inhibited by GSH. Hence, quenching of fluorescence is prevented. This effect was used to design a fluorometric assay for GSH that has good selectivity and sensitivity. Under optimal conditions, the method has a linear response in the 0.2 to 20 μM GSH concentration range and a 60 nM detection limit. It was successfully applied to the determination of GSH in HepG2 cells and in spiked human serum.

Schematic representation of using a NIR fluorescent P-dots and dopamine (DA)-melanin nanohybrid as a probe for glutathione (GSH) detection. The P-dots were prepared from poly(styrene-co-maleic anhydride) (PSMA), the semiconducting polymer poly[(9,9′-dioctyl-2,7-divinylenefluorenylene)-alt-2-methoxy-5-(2-ethyl-hexyloxy)-1,4-phenylene] (PEPV) and the fluorescent dye tetraphenylporphyrin (TPP).The GSH can inhibit the dopamine self-polymerization and prevented the formation of PDA and fluorescence quenching of P-dots.

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References

  1. 1.

    Valko M, Rhodes CJ, Moncol J et al (2006) Free radicals, metals and antioxidants in oxidative stress-induced cancer. Chem Biol Interact 160:1–40. https://doi.org/10.1016/j.cbi.2005.12.009

    CAS  Article  PubMed  Google Scholar 

  2. 2.

    Pastore A, Federici G, Bertini E, Piemonte F (2003) Analysis of glutathione: implication in redox and detoxification. Clin Chim Acta 333:19–39. https://doi.org/10.1016/S0009-8981(03)00200-6

    CAS  Article  PubMed  Google Scholar 

  3. 3.

    Sies H (1999) Glutathione and its role in cellular functions. Free Radic Biol Med 27:916–921. https://doi.org/10.1016/S0891-5849(99)00177-X

    CAS  Article  PubMed  Google Scholar 

  4. 4.

    Dröge W, Breitkreutz R (2000) Glutathione and immune function. Proc Nutr Soc 59:595–600. https://doi.org/10.1017/S0029665100000847

    Article  PubMed  Google Scholar 

  5. 5.

    Estrela JM, Ortega A, Obrador E (2006) Glutathione in Cancer Biology and Therapy. Crit Rev Clin Lab Sci 43:143–181. https://doi.org/10.1080/10408360500523878

    CAS  Article  PubMed  Google Scholar 

  6. 6.

    Ballatori N, Krance SM, Notenboom S et al (2009) Glutathione dysregulation and the etiology and progression of human diseases. Biol Chem 390:191–214. https://doi.org/10.1515/BC.2009.033

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  7. 7.

    Wu G, Fang Y-Z, Yang S et al (2004) Glutathione Metabolism and Its Implications for Health. J Nutr 134:489–492. https://doi.org/10.1093/jn/134.3.489

    CAS  Article  PubMed  Google Scholar 

  8. 8.

    McDermott GP, Francis PS, Holt KJ et al (2011) Determination of intracellular glutathione and glutathione disulfide using high performance liquid chromatography with acidic potassium permanganate chemiluminescence detection. Analyst 136:2578–2585. https://doi.org/10.1039/c1an00004g

    CAS  Article  PubMed  Google Scholar 

  9. 9.

    Lee PT, Goncalves LM, Compton RG (2015) Electrochemical determination of free and total glutathione in human saliva samples. Sensors Actuators B Chem 221:962–968. https://doi.org/10.1016/j.snb.2015.07.050

    CAS  Article  Google Scholar 

  10. 10.

    Feng J, Huang P, Shi S et al (2017) Colorimetric detection of glutathione in cells based on peroxidase-like activity of gold nanoclusters: A promising powerful tool for identifying cancer cells. Anal Chim Acta 967:64–69. https://doi.org/10.1016/j.aca.2017.02.025

    CAS  Article  PubMed  Google Scholar 

  11. 11.

    Liu J, Meng L, Fei Z et al (2017) MnO2 nanosheets as an artificial enzyme to mimic oxidase for rapid and sensitive detection of glutathione. Biosens Bioelectron 90:69–74. https://doi.org/10.1016/j.bios.2016.11.046

    CAS  Article  PubMed  Google Scholar 

  12. 12.

    Xu H-H, Deng H-H, Lin X-Q et al (2017) Colorimetric glutathione assay based on the peroxidase-like activity of a nanocomposite consisting of platinum nanoparticles and graphene oxide. Microchim Acta 184:3945–3951. https://doi.org/10.1007/s00604-017-2429-3

    CAS  Article  Google Scholar 

  13. 13.

    Gao W, Liu Z, Qi L et al (2016) Ultrasensitive Glutathione Detection Based on Lucigenin Cathodic Electrochemiluminescence in the Presence of MnO2 Nanosheets. Anal Chem 88:7654–7659. https://doi.org/10.1021/acs.analchem.6b01491

    CAS  Article  PubMed  Google Scholar 

  14. 14.

    Wei C, Liu X, Gao Y et al (2018) Thiol–Disulfide Exchange Reaction for Cellular Glutathione Detection with Surface-Enhanced Raman Scattering. Anal Chem 90:11333–11339. https://doi.org/10.1021/acs.analchem.8b01974

    CAS  Article  PubMed  Google Scholar 

  15. 15.

    Zhang X-L, Zheng C, Guo S-S et al (2014) Turn-On Fluorescence Sensor for Intracellular Imaging of Glutathione Using g-C3N4 Nanosheet–MnO2 Sandwich Nanocomposite. Anal Chem 86:3426–3434. https://doi.org/10.1021/ac500336f

    CAS  Article  PubMed  Google Scholar 

  16. 16.

    Zhang D, Zheng A, Li J, et al (2017) Tumor microenvironment activable self-assembled DNA hybrids for pH and redox dual-responsive chemotherapy/PDT treatment of hepatocellular carcinoma. Adv Sci 1600460. https://doi.org/10.1002/advs.201600460

  17. 17.

    Yang Y, Zhao Q, Feng W, Li F (2013) Luminescent Chemodosimeters for Bioimaging. Chem Rev 113:192–270. https://doi.org/10.1021/cr2004103

    CAS  Article  PubMed  Google Scholar 

  18. 18.

    Niu L-Y, Guan Y-S, Chen Y-Z et al (2012) BODIPY-Based Ratiometric Fluorescent Sensor for Highly Selective Detection of Glutathione over Cysteine and Homocysteine. J Am Chem Soc 134:18928–18931. https://doi.org/10.1021/ja309079f

    CAS  Article  PubMed  Google Scholar 

  19. 19.

    Liu J, Bao C, Zhong X et al (2010) Highly selective detection of glutathione using a quantum-dot-based OFF-ON fluorescent probe. Chem Commun (Camb) 46:2971–2973. https://doi.org/10.1039/b924299f

    CAS  Article  Google Scholar 

  20. 20.

    Tian D, Qian Z, Xia Y, Zhu C (2012) Gold Nanocluster-Based Fluorescent Probes for Near-Infrared and Turn-On Sensing of Glutathione in Living Cells. Langmuir 28:3945–3951. https://doi.org/10.1021/la204380a

    CAS  Article  PubMed  Google Scholar 

  21. 21.

    Zhou L, Lin Y, Huang Z et al (2012) Carbon nanodots as fluorescence probes for rapid, sensitive, and label-free detection of Hg2+ and biothiols in complex matrices. Chem Commun 48:1147–1149. https://doi.org/10.1039/C2CC16791C

    CAS  Article  Google Scholar 

  22. 22.

    Deng R, Xie X, Vendrell M et al (2011) Intracellular Glutathione Detection Using MnO2 -Nanosheet-Modified Upconversion Nanoparticles. J Am Chem Soc 133:20168–20171. https://doi.org/10.1021/ja2100774

    CAS  Article  PubMed  Google Scholar 

  23. 23.

    Feng L, Zhu C, Yuan H et al (2013) Conjugated polymer nanoparticles: preparation, properties, functionalization and biological applications. Chem Soc Rev 42:6620–6633. https://doi.org/10.1039/c3cs60036j

    CAS  Article  PubMed  Google Scholar 

  24. 24.

    Wu C, Chiu DT (2013) Highly Fluorescent Semiconducting Polymer Dots for Biology and Medicine. Angew Chem Int Ed 52:3086–3109. https://doi.org/10.1002/anie.201205133

    CAS  Article  Google Scholar 

  25. 25.

    Lyu Y, Pu K (2017) Recent Advances of Activatable Molecular Probes Based on Semiconducting Polymer Nanoparticles in Sensing and Imaging. Adv Sci 4:1600481. https://doi.org/10.1002/advs.201600481

    CAS  Article  Google Scholar 

  26. 26.

    Guo L, Niu G, Zheng X et al (2017) Single Near-Infrared Emissive Polymer Nanoparticles as Versatile Phototheranostics. Adv Sci 4:1700085. https://doi.org/10.1002/advs.201700085

    CAS  Article  Google Scholar 

  27. 27.

    Yin C, Zhen X, Fan Q et al (2017) Degradable Semiconducting Oligomer Amphiphile for Ratiometric Photoacoustic Imaging of Hypochlorite. ACS Nano 11:4174–4182. https://doi.org/10.1021/acsnano.7b01092

    CAS  Article  PubMed  Google Scholar 

  28. 28.

    Sun K, Tang Y, Li Q et al (2016) In Vivo Dynamic Monitoring of Small Molecules with Implantable Polymer-Dot Transducer. ACS Nano 10:6769–6781. https://doi.org/10.1021/acsnano.6b02386

    CAS  Article  PubMed  Google Scholar 

  29. 29.

    Wang D, Chen C, Ke X et al (2015) Bioinspired Near-Infrared-Excited Sensing Platform for in Vitro Antioxidant Capacity Assay Based on Upconversion Nanoparticles and a Dopamine–Melanin Hybrid System. ACS Appl Mater Interfaces 7:3030–3040. https://doi.org/10.1021/am5086269

    CAS  Article  PubMed  Google Scholar 

  30. 30.

    Matsuki M, Watanabe T, Ogasawara A et al (2008) Inhibitory Mechanism of Melanin Synthesis by Glutathione. Yakugaku Zasshi 128:1203–1207. https://doi.org/10.1248/yakushi.128.1203

    CAS  Article  PubMed  Google Scholar 

  31. 31.

    Ahsan H (2011) Arsenic in drinking water. Kaohsiung J Med Sci 27:358–359. https://doi.org/10.1016/j.kjms.2011.05.002

    Article  PubMed  Google Scholar 

  32. 32.

    Saha A, Jana NR (2013) Detection of Cellular Glutathione and Oxidized Glutathione Using Magnetic–Plasmonic Nanocomposite-Based “Turn-Off” Surface Enhanced Raman Scattering. Anal Chem 85:9221–9228. https://doi.org/10.1021/ac4019457

    CAS  Article  PubMed  Google Scholar 

  33. 33.

    Yu F, Li P, Wang B, Han K (2013) Reversible Near-Infrared Fluorescent Probe Introducing Tellurium to Mimetic Glutathione Peroxidase for Monitoring the Redox Cycles between Peroxynitrite and Glutathione in Vivo. J Am Chem Soc 135:7674–7680. https://doi.org/10.1021/ja401360a

    CAS  Article  PubMed  Google Scholar 

  34. 34.

    Wang Y, Jiang K, Zhu J et al (2015) A FRET-based carbon dot–MnO2 nanosheet architecture for glutathione sensing in human whole blood samples. Chem Commun 51:12748–12751. https://doi.org/10.1039/C5CC04905A

    CAS  Article  Google Scholar 

Download references

Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grant No. 21605021 and 21705022), Joint Funds for the Innovation of Science and Technology of Fujian province (Grant No. 2016Y9060 and 2017Y9115), the Young and Middle-aged Talent Training Project of Fujian Provincial Health and Family Planning Commission (Grant No. 2018-ZQN-75), the China Postdoctoral Science Foundation, the Natural Science Foundation of Fujian Province of China (Grant No. 2016 J05206), the Medical Innovation grant of Fujian province (Grant No. 2018-CX-49) and the Startup Fund of Mengchao Hepatobiliary Hospital of Fujian Medical University (Grant No. QDZJ-2017-004).

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  1. Jing Wang and Cheng Zheng contributed equally to this work.

Affiliations

  1. The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi’an Jiaotong University, Xi’an, 710049, People’s Republic of China

    Jing Wang, Zhenxi Zhang & Xiaolong Liu

  2. College of Chemistry, Fuzhou University, Fuzhou, 350116, People’s Republic of China

    Cheng Zheng

  3. The United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou, 350025, People’s Republic of China

    Xionghong Tan, Aixian Zheng, Yongyi Zeng, Xiaolong Zhang & Xiaolong Liu

  4. The Liver Center of Fujian Province, Fujian Medical University, Fuzhou, 350025, People’s Republic of China

    Xionghong Tan, Aixian Zheng, Yongyi Zeng, Xiaolong Zhang & Xiaolong Liu

  5. Liver Disease Center, The First Affiliated Hospital of Fujian Medical University, Fuzhou, 350005, People’s Republic of China

    Yongyi Zeng

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  1. Jing Wang
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Wang, J., Zheng, C., Tan, X. et al. Sensitive fluorometric determination of glutathione using fluorescent polymer dots and the dopamine-melanin nanosystem. Microchim Acta 186, 568 (2019). https://doi.org/10.1007/s00604-019-3675-3

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Keywords

  • Polydopamine
  • Semiconducting polymer dots
  • Antiquenching
  • Antioxidant
  • PEPV
  • Biothiols
  • Tetraphenylporphyrin
  • Cell extracts
  • Serum
  • Nanoparticles