Development of an Ultrasensitive Mitochondria-Targeted Near Infrared Fluorescent Probe for SO2 and Its Imaging in Living Cells and Mice

doi:10.6023/cjoc202012049

Chinese Journal of Organic Chemistry ›› 2021, Vol. 41 ›› Issue (3): 1108-1116.DOI: 10.6023/cjoc202012049 Previous Articles Next Articles

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高灵敏线粒体靶向近红外二氧化硫荧光探针的开发及细胞、小鼠成像研究

李芳¹, 唐永和¹, 郭锐¹, 林伟英¹^,^*()

1 广西电化学能源材料重点实验室广西大学光功能材料与化学生物学研究院广西大学化学化工学院有色金属及材料加工新技术教育部重点实验室广西有色金属及特色材料加工重点实验室南宁 530004

收稿日期:2020-12-29 修回日期:2021-02-14 发布日期:2021-02-26
通讯作者: 林伟英
基金资助:
国家自然科学基金(21672083); 国家自然科学基金(21877048); 国家自然科学基金(22077048); 广西大学启动基金(A3040051003); 广西自然科学基金(2019GXNSFBA245068)

Development of an Ultrasensitive Mitochondria-Targeted Near Infrared Fluorescent Probe for SO₂ and Its Imaging in Living Cells and Mice

Fang Li¹, Yonghe Tang¹, Rui Guo¹, Weiying Lin¹^,^*()

1 Guangxi Key Laboratory of Electrochemical Energy Materials, Institute of Optical Materials and Chemical Biology, School of Chemistry and Chemical Engineering, MOE Key Laboratory of New Processing Technology for Nonferrous Metals and Materials, Guangxi Key Laboratory of Processing for Non-ferrous Metals and Featured Materials, Guangxi University, Nanning 530004

Received:2020-12-29 Revised:2021-02-14 Published:2021-02-26
Contact: Weiying Lin
About author:
* Corresponding author. E-mail: weiyinglin2013@163.com
Supported by:
Project supported by the NSFC(21672083); Project supported by the NSFC(21877048); Project supported by the NSFC(22077048); startup fund of Guangxi University(A3040051003); Guangxi Natural Science Foundation(2019GXNSFBA245068)

1. 1614331807470-1818850272辅助材料.pdf(2085KB)

Abstract

Sulfur dioxide (SO₂) and its derivatives (sulfites, bisulfites, etc.), as important components of active sulfur species, are closely related to the physiological processes of living organisms. In addition, excessive intake of SO₂ may cause irreparable damage to the respiratory system, such as lung cancer, tracheitis, asthma, etc. Near-infrared emission fluorescent probes have strong penetrating ability and can detect biomolecules in vivoeffectively. Therefore, in this paper, a mitochondrial targeted near-infrared SO₂ fluorescent probe based on the Michael addition mechanism was designed. The solution spectroscopic experiments show that XA-SO₂ has significant selectivity and sensitivity to SO₂, and has the ability to rapidly detect SO₂. It is worth noting that XA-SO₂ has good membrane permeability and has successfully detected extracellular/endogenous levels. More importantly, XA-SO₂ could be enriched effectively on the mitochondria of cells, which is promising for realizing the dynamic detection of SO₂ in the mitochondria of living cells.

Key words: sulfur dioxide, fluorescent probe, bioimaging, Michael addition, mitochondria

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Fang Li, Yonghe Tang, Rui Guo, Weiying Lin. Development of an Ultrasensitive Mitochondria-Targeted Near Infrared Fluorescent Probe for SO₂ and Its Imaging in Living Cells and Mice[J]. Chinese Journal of Organic Chemistry, 2021, 41(3): 1108-1116.

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Fig. & Tab. 11

Scheme 1. Proposed response mechanism of probe XA-SO2 to SO2

Scheme 2. Synthetic route of probe XA-SO2

Figure 1. UV-Vis absorption curves of probe XA-SO2 (5×10–6 mol/L) reacted with different concentrations of NaHSO3 (0~8×10–4 mol/L) UV-Vis absorption curves of XA-SO2 (5×10–6 mol/L) reacted with different concentrations of NaHSO3 (0~8×10–4 mol/L) in 1×10–2 mol/L PBS buffer (pH 7.4, 0.5% DMSO) for 10 min

Figure 2. Fluorescent curves of probe XA-SO2 (5×10–6 mol/L) reacted with different concentrations of NaHSO3 (0~8×10–4 mol/L) Fluorescent curves of XA-SO2 (5×10–6 mol/L) reacted with different concentrations of NaHSO3 (1×10–6~1.5×10–5 mol/L) in 1×10–2 mol/L PBS buffer (pH 7.4, 0.5% DMSO) for 10 min. λ ex=670 nm

Figure 3. Linear relationship between the fluorescence intensity of the probe XA-SO2 and the concentration of NaHSO3 (1×10–6~1.5×10–5 mol/L) at 755 nm Linear relationship between the fluorescence intensity of the probe XA-SO2 (5×10–6 mol/L) and the concentration of NaHSO3 (1×10–6~1.5×10–5 mol/L) at 755 nm

Figure 4. Fluorescence intensity (755 nm) curve of probe XA-SO2 in the presence SO2 at different time (0~60 s). Time-dependent fluorescent intensities (F755 nm) of probe XA-SO2 (5×10–6 mol/L) in the presence SO2

Figure 5. Fluorescence intensity of the reaction system of probe 5×10–6 mol/L XA-SO2 with various analytes at 755 nm The numbers represent: (1) probe, (2) glucose, (3) DL-Hcy, (4) H2O2, (5) FA, (6) L-Cys, (7) L-Ala, (8) L-Arg, (9) Ascorbic Acid, (10) NaClO, (11) NaHS, (12) KI, (13) NaF, (14) Gly, (15) GSH, (16) Na2S, (17) NaSCN, (18) Na2SO4, (19) TBHP, (20) KCl, (21) MgCl2, (22) CaCl2, (23) SNP, (24) NaNO2, (25) Na2SO3, (26) NaHSO3. λ ex=670 nm

Figure 6. Fluorescence intensity of probe XA-SO2 and SO2 in different pH solutions at 755 nm The fluorescence intensity changes of XA-SO2 (5×10–6 mol/L) treated or untreated with SO2 (1.2×10–5 mol/L) at different pH (6.0~10.0) solutions for 10 min

Figure 7. Fluorescent images of probe XA-SO2 in HeLa cells for detecting added NaHSO3 (A) The HeLa cells were incubated with 1×10–6 mol/L probe XA-SO2 for 5 min; (B) The HeLa cells were pre-cultured with 1×10–5 mol/L NaHSO3 for 5 min, and then cultured with 1×10–6 mol/L probe XA-SO2 for another 5 min; (C) The HeLa cells were pre-cultured with 1×10–4 mol/L NaHSO3 for 5 min, and then cultured with 1×10–6 mol/L probe XA-SO2 for another 5 min. λ ex=640 nm, λ em=650~750 nm, scale bar=30 μm

Figure 8. Fluorescent images of probe XA-SO2 in HeLa cells for detecting endogenous SO2 (A) The HeLa cells were incubated with 1×10–6 mol/L probe XA-SO2 for 5 min; (B) The HeLa cells were pre-cultured with 2×10–5 mol/L Cys for 20 min, and then cultured with 1×10–6 mol/L probe XA-SO2 for another 5 min; (C) The HeLa cells were pre-cultured with 5×10–5mol/L Cys for 20 min, and then cultured with 1×10–6 mol/L probe XA-SO2 for another 5 min. λ ex=640 nm, λ em=650~750 nm, scale bar=30 μm

Figure 9. Co-location imaging experiment of probe XA-SO2 and commercial dye (Mito Tracker Green) Co-location imaging experiments between probe XA-SO2 and Mito Tracker Green. The green channel: λ ex=488 nm, λ em=500~550 nm; The red channel: λ ex=640 nm, λ em=650~750 nm. Scale bar: 30 μm

References 42

[1]	Sun, Y. Q.; Liu, J.; Zhang, J.; Yang, T.; Guo, W. Chem. Commun. 2013, 49, 2637.
[2]	Meng, Z.; Yang, Z.; Li, J.; Zhang, Q. Chemosphere 2012, 89, 579. pmid: 22763331
[3]	Smith, T.; Peters, J.; Reading, J.; Castle, C. Am. Rev. Respir. Dis. 1977, 116, 31. pmid: 879598
[4]	Cheng, W.; Xu, J.; Guo, Z.; Yang, D.; Chen, X.; Yan, W.; Miao, P. J. Mater. Chem. B 2018, 6, 5775.
[5]	Kobayashi, S.; Tsuzuki, M.; Sato, N. Plant Cell Physiol. 2015, 56, 1521. pmid: 26009593
[6]	Beeson, W.; Abbey, D.; Knutsen, S. Environ. Health Perspect. 1998, 106, 813. doi: 10.1289/ehp.98106813 pmid: 9646042
[7]	Zhang, Q.; Prabhu, A.; San, A.; Al-Sharab, J.; Levon, K. Biosens. Bioelectron. 2015, 72, 100. pmid: 25966464
[8]	Lee, W.; Teschke, K.; Kauppinen, T.; Andersen, A.; Jappinen, P.; Szadkowskastanczyk, I.; Pearce, N.; Persson, B.; Environ, A. Health Perspect. 2002, 110, 991.
[9]	Sang, N.; Yun, Y.; Yao, G. Y.; Li, H. Y.; Guo, L.; Li, G. K. Toxicol. Sci. 2011, 124, 400. doi: 10.1093/toxsci/kfr224 pmid: 21873648
[10]	Stipanuk, M.; Ueki, I. J. Inherited Metab. Dis. 2011, 34, 17. pmid: 20162368
[11]	Ubuka, T.; Ohta, J.; Yao, W.; Abe, T.; Teraoka, T.; Kurozumi, Y. Amino Acids 1992, 2, 143. doi: 10.1007/BF00806085 pmid: 24194282
[12]	Li, J.; Meng, Z.; Nitric Oxide, 2009, 20, 166. pmid: 19135162
[13]	Jin, H. F.; Wang, Y.; Wang, X. B.; Sun, Y.; Tang, C. S.; Du, J. B. Nitric Oxide 2013, 32, 561.
[14]	Wang, X. B.; Huang, X. M.; Ochs, T.; Li, X. Y.; Jin, H. F.; Tang, C. S.; Du, J. B. Basic Res. Cardiol. 2011, 106, 865. pmid: 21468766
[15]	Meng, Z.; Yang, Z.; Li, J.; Zhang, Q. Chemosphere 2012, 89, 579. pmid: 22763331
[16]	Wang, X. B.; Du, J. B.; Cui, H. Life Sci. 2014, 98, 63. pmid: 24412383
[17]	Zorov, D. B.; Isaev, N. K.; Plotnikov, E. Y.; Zorova, L. D.; Stelmashook, E. V.; Vasileva, A. K.; Arkhangelskaya, A. A.; Khrjapenkova, T. G. Biochemistry 2007, 72, 1115. pmid: 18021069
[18]	Du, S. X.; Jin, H. X.; Bu, D. F.; Zhao, X.; Geng, B.; Tang, C. S.; Du, J. B. Acta Pharmacol. Sin. 2008, 29, 923. pmid: 18664325
[19]	Xu, W.; Teoh, C. L.; Peng, J.; Su, D.; Yuan, L.; Chang, Y.-T. Biomaterials 2015, 56, 1. pmid: 25934273
[20]	Zhao, Y.; Qiu, W.; Yang, C.; Wang, J. Energy Fuels 2017, 31, 693.
[21]	Palenzuela, B.; Simonet, B.; Rı´os, A.; Valcárcel, M. Anal. Chim. Acta 2005, 535, 65.
[22]	Daunoravicius, Z.; Padarauskas, A. Electrophoresis 2002, 23, 2439. pmid: 12210200
[23]	Pundir, C.; Rawal, R. Anal. Bioanal. Chem. 2013, 405, 3049. pmid: 23392406
[24]	Segundo, M.; Rangel, A.; Cladera, A.; Cerdá, V. Analyst 2000, 125, 1501. doi: 10.1039/b002225j
[25]	Williams, T.; McElvany, S.; Ighodalo, E. Anal. Chim. Acta 1981, 123, 351. doi: 10.1016/S0003-2670(01)83194-9
[26]	Yue, Y. K.; Huo, F. J.; Yin, C. X. Sci. China Chem. 2017, 42, 249. (in Chinese)
	(岳永康, 霍方俊, 阴彩霞, 中国科学: 化学, 2017, 42, 249.)
[27]	Zhang, S. X.; Niu, Q. M.; Wu, S. Z.; Lu, H. J.; Xing, G. W. Chin. J. Org. Chem. 2019, 39, 940. (in Chinese)
	(张晟曦, 牛晴旻, 吴松泽, 吕海娟, 邢国文, 有机化学, 2019, 39, 940.)
[28]	Zhang, J. D.; Liu, A. C.; Chen, J.; Yuan, G. H.; Jin, H. Y. Prog. Chem. 2020, 32, 594. (in Chinese)
	(张继东, 刘阿晨, 陈娇, 袁光辉, 金华峰, 化学进展, 2020, 32, 594.)
[29]	Zhang, L.; Li, X. A.; Gillis, K. D.; Glass, T. E. Angew. Chem. Int. Ed. 2019, 58, 7611. doi: 10.1002/anie.v58.23
[30]	Yue, Y. K.; Huo, F. G.; Ning, P.; Zhang, Y. B.; Chao, J. B.; Meng, X. M.; Yin, C. X. J. Am. Chem. Soc. 2017, 139, 3181. pmid: 28170238
[31]	Cao, D. X.; Liu, Z. Q.; Verwilst, P.; Koo, S.; Jangjili, P.; Kim, J. S.; Lin, W. Y. Chem. Rev. 2019, 119, 10403. pmid: 31314507
[32]	Yang, B.; Xua, J.; Zhu, H. L. Free Radical Biol. Med. 2019, 145, 42.
[33]	Jia, L.; Niu, L, Y.; Yang, Q, Z. Anal. Chem. 2020, 92, 10800. doi: 10.1021/acs.analchem.0c02255 pmid: 32605361
[34]	Li, J. Z.; Sun, Y. H.; Wang, C. Y.; Cao, S. Y.; Guo, Z. Q.; Shen, Y. J.; Zhu, W. H. Anal. Chem. 2019, 91, 11946. pmid: 31423770
[35]	Li, T.; Huo, F. J.; Chao, J. B.; Yin, C. X. Chem. Commun. 2020, 56, 11453.
[36]	Chen, H.; Dong, B. L.; Tang, Y. H.; Lin, W. Y. Acc. Chem. Res. 2017, 50, 6, 1410. pmid: 28492303
[37]	Zhang, W. J.; Liu, T.; Huo, F. J.; Ning, P.; Wen, Y.; Meng, X. M.; Yin, C. X. Sci. China Chem. 2017, 47, 1022. (in Chinese)
	(张伟杰, 刘涛, 霍方俊, 温莹, 孟祥明, 阴彩霞, 中国科学: 化学, 2017, 47, 1022.)
[38]	Zhang, W. J.; Huo, F, J.; Cheng, F, Q.; Yin, C. Y. J. Am. Chem. Soc. 2020, 142, 13, 6324. pmid: 32130860
[39]	Geng, L. H.; Yang, X. F.; Zhong, Y. G.; Li, Z.; Li, H. Dyes Pigm. 2015, 120, 213.
[40]	Huang, C, B.; Chen, H.; Li, F. Q.; An, S. Y.; Chin. J. Org. Chem. 2019, 39, 2467. (in Chinese)
	(黄池宝, 陈会, 李福琴, 安思雅, 有机化学, 2019, 39, 2467.)
[41]	Yan, Y. H.; Wu, Q. R.; Che, Q. L.; Ding, M. M.; Xu, M.; Miao, J. Y.; Zhao, B. X.; Lin, Z. M. Analyst 2020, 145, 2937. pmid: 32104823
[42]	Zhang, W. J.; Huo, F. J.; Zhang, Y. B.; Yin, C. X. J. Mater. Chem. B 2019, 7, 1945.

高灵敏线粒体靶向近红外二氧化硫荧光探针的开发及细胞、小鼠成像研究

Development of an Ultrasensitive Mitochondria-Targeted Near Infrared Fluorescent Probe for SO₂ and Its Imaging in Living Cells and Mice

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