Citation: Lu Chang, Junjie Li, Jing Zhang, Fang Yuan, Liang-Liang Gao, Ji-Wei Shen, Jianjian Zhang, Yuan Guo. Lysosome-targeted ratiometric fluorescent probe for dynamic assessment of nitric oxide in cellular senescence[J]. Chinese Chemical Letters, ;2026, 37(7): 111745. doi: 10.1016/j.cclet.2025.111745 shu

Lysosome-targeted ratiometric fluorescent probe for dynamic assessment of nitric oxide in cellular senescence

Engineering Research Center of Western Resource Innovation Medicine Green Manufacturing of the Ministry of Education, School of Chemical Engineering, Key Laboratory of Synthetic and Natural Functional Molecule of the Ministry of Education, College of Chemistry and Materials Science, Northwest University, Xi'an 710127, China

jiweish@nwu.edu.cn

zhangjj@nwu.edu.cn

guoyuan@nwu.edu.cn

Received Date: 16 June 2025
Revised Date: 18 August 2025
Accepted Date: 19 August 2025
Available Online: 20 August 2025

Figures(5)

Nitric oxide (NO), a significant signaling molecule, plays essential roles in diverse physiological and pathological processes including cell senescence. Lysosomes are recognized as pivotal subcellular organelles of cellular senescence, whereas the influence of senescence on lysosomal NO homeostasis remains poorly understood. To explore the potential association between cellular senescence and alterations in lysosomal NO levels, herein, we developed FL-O1, a novel ratiometric fluorescent probe specifically designed for lysosomal NO detection. The probe selectively reacts with NO through the recognition unit of o-phenylenediamine, forming an extended conjugated system that generates a distinct ratiometric optical signal with exceptional sensitivity (detection limit = 95 nmol/L). FL-O1 exhibits versatile applications in both in vitro and in vivo contexts. It successfully visualizes exogenous and endogenous NO dynamics in live zebrafish models and enables specific monitoring of lysosomal NO levels in cellular systems. Importantly, the probe was utilized to quantitatively evaluate NO levels in senescent states of cancer and healthy cells. The results revealed two key insights: (1) Senescent cells consistently maintain higher NO concentrations compared to their non-senescent counterparts, and (2) senescent healthy cells exhibit elevated NO levels relative to senescent cancer cells. This innovative small-molecule probe represents a significant advancement in lysosomal NO imaging technology. Its unique design and capabilities provide a powerful tool for investigating the role of lysosomal NO in cellular senescence, potentially uncovering novel mechanistic insights and facilitating the development of targeted therapeutic strategies for senescence-related disorders.
- Nitric oxide detection,
- Ratiometric,
- Fluorescent probe,
- Lysosome-targeted,
- Cellular senescence
1. [1]
  J. Lundberg, J. Am. Coll. Cardiol. 47 (2006) 580–581. doi: 10.1016/j.jacc.2005.11.016
2. [2]
  W. Jiang, W. Dong, M. Li, et al., ACS Nano. 16 (2022) 3881–3894. doi: 10.1021/acsnano.1c09048
3. [3]
  D. Bredt, P. Hwang, S. Snyder, Nature 347 (1990) 768–770. doi: 10.1038/347768a0
4. [4]
  W. Xu, L. Liu, M. Loizidou, et al., Cell Res. 12 (2002) 311–320. doi: 10.1038/sj.cr.7290133
5. [5]
  R.B. Salimian, A. Achreja, D. Nagrath, Trends Cancer. 3 (2017) 659–672. doi: 10.1016/j.trecan.2017.07.005
6. [6]
  M.S. Finkel, C.V. Oddis, T.D. Jacob, et al., Science 257 (1992) 387–389. doi: 10.1126/science.1631560
7. [7]
  J. Lundberg, E. Weitzberg, Cell 185 (2022) 2853–2878. doi: 10.1016/j.cell.2022.06.010
8. [8]
  K. Chachlaki, J. Garthwaite, V. Prevot, Nat. Rev. Endocrinol. 13 (2017) 521–535. doi: 10.1038/nrendo.2017.69
9. [9]
  T. Nakamura, C.K. Oh, L. Liao, Science 371 (2021) eaaw0843. doi: 10.1126/science.aaw0843
10. [10]
  D. Tewari, A.N. Sah, S. Bawari, et al., Curr. Neuropharmacol. 19 (2021) 114–126. doi: 10.2174/18756190mta2pmjapy
11. [11]
  Z. Wu, J. Qu, G.H. Liu, Nat. Rev. Mol. Cell Biol. 25 (2024) 979–1000. doi: 10.1038/s41580-024-00775-3
12. [12]
  H.A. Shaban, S.M. Gasser, Cell Death Differ. 32 (2025) 9–15. doi: 10.1038/s41418-023-01197-y
13. [13]
  L. Hayflick, P.S. Moorhead, Exp. Cell Res. 25 (1961) 585–621. doi: 10.1016/0014-4827(61)90192-6
14. [14]
  J.C. Wang, J. Wei, H. Inuzuka, Acta Mater. Med. 2 (2023) 281–284. doi: 10.3390/land12020281
15. [15]
  A. Jaiswal, N. Kaushik, T.R. Acharya, et al., ACS Appl. Mater. 16 (2024) 59991–60007. doi: 10.1021/acsami.4c14051
16. [16]
  T. Hayashi, K. Yano, H. Matsui-Hirai, et al., Pharmacol. Ther. 120 (2008) 333–339. doi: 10.1016/j.pharmthera.2008.09.002
17. [17]
  A.M. Pourbagher-Shahri, T. Farkhondeh, M. Talebi, et al., Molecules 26 (2021) 4533. doi: 10.3390/molecules26154533
18. [18]
  N. Mabrouk, S. Ghione, V. Laurens, et al., Cancers 12 (2020) 1145. doi: 10.3390/cancers12051145
19. [19]
  Q. Jiang, G. Song, L. He, et al., Kidney Int. 106 (2024) 419–432. doi: 10.1016/j.kint.2024.05.007
20. [20]
  B.H. Min, H.J. Kim, H. Lim, et al., Exp. Mol. Med. 33 (2001) 299–302. doi: 10.1038/emm.2001.48
21. [21]
  J.A. Fafián-Labora, A. O'Loghlen, Aging Cell 20 (2021) e13426. doi: 10.1111/acel.13426
22. [22]
  B. Xu, Y. Liu, N. Li, Front. Immunol. 15 (2024) 1395786. doi: 10.3389/fimmu.2024.1395786
23. [23]
  G. Morris, M. Gevezova, V. Sarafian, et al., Cell Mol. Immunol. 19 (2022) 1079–1101. doi: 10.1038/s41423-022-00902-0
24. [24]
  Y.M. Janssen-Heininger, M.E. Poynter, P.A. Baeuerle, Free Radical Biol. Med. 28 (2000) 1317–1327. doi: 10.1016/S0891-5849(00)00218-5
25. [25]
  P. Pacher, J.S. Beckman, L. Liaudet, Physiol. Rev. 87 (2007) 315–424. doi: 10.1152/physrev.00029.2006
26. [26]
  A.N. Blackford, S.P. Jackson, Mol. Cell 66 (2017) 801–817. doi: 10.1016/j.molcel.2017.05.015
27. [27]
  I. Sturmlechner, M. Durik, C.J. Sieben, et al., Nat. Rev. Nephrol. 13 (2016) 77–89.
28. [28]
  X. Qu, P.Z. Wu, K.X. Duan, et al., Chin. Chem. Lett. 35 (2024) 109681. doi: 10.1016/j.cclet.2024.109681
29. [29]
  K. Roh, J. Noh, Y. Kim, et al., Nat. Metab. 5 (2023) 398–413. doi: 10.1038/s42255-023-00747-5
30. [30]
  L.P. Fang, C.H. Lin, Y. Medlej, et al., Nat. Commun. 16 (2025) 1175. doi: 10.1038/s41467-025-56484-8
31. [31]
  W. Wang, T. Yang, Q. Zhang, et al., Anal. Chem. 95 (2023) 6279–6286. doi: 10.1021/acs.analchem.2c04896
32. [32]
  X.Y. Wang, Y.M. Lin, X.Y. Sun, et al., Sens. Actuators B: Chem. 383 (2023) 133592. doi: 10.1016/j.snb.2023.133592
33. [33]
  Z. Xu, S. Liu, L. Xu, et al., Anal. Chim. Acta 1297 (2024) 342303. doi: 10.1016/j.aca.2024.342303
34. [34]
  D. Andina, J.C. Leroux, P. Luciani, Chem. Eur. J. 23 (2017) 13549–13573. doi: 10.1002/chem.201702458
35. [35]
  R. Huang, Z. Zhang, Z. Shi, et al., Chem. Commun. 60 (2024) 6793–6796. doi: 10.1039/d4cc02133a
36. [36]
  S.L. Xu, F.F. Guo, Z.H. Xu, et al., Sens. Actuators B: Chem. 383 (2023) 133510. doi: 10.1016/j.snb.2023.133510
37. [37]
  S. Munan, A. Mondal, S. Shailja, et al., Anal. Chem. 96 (2024) 7479–7486. doi: 10.1021/acs.analchem.3c05981
38. [38]
  M. Liu, B. Feng, F. Chu, et al., Chin. Chem. Lett. 36 (2025) 110043. doi: 10.1016/j.cclet.2024.110043
39. [39]
  J. Tian, D. Shi, Y. Zhang, et al., Chem. Sci. 12 (2021) 13483–13491. doi: 10.1039/d1sc04162b
40. [40]
  L. Zeng, Y. Ke, X. Yang, et al., Food Chem. 438 (2024) 138044. doi: 10.1016/j.foodchem.2023.138044
41. [41]
  Y. Li, K. Pan, Y. Gao, et al., Chem. Sci. 15 (2024) 6562–6571. doi: 10.1039/d4sc00767k
42. [42]
  N.N. Atia, P.Y. Khashaba, S.A. El Zohny, et al., Talanta 272 (2024) 125763. doi: 10.1016/j.talanta.2024.125763
43. [43]
  J. Hu, R. Wang, W. Liao, et al., Anal. Chim. Acta 1296 (2024) 342333. doi: 10.1016/j.aca.2024.342333
44. [44]
  Z. Zeng, S. Gong, Q. Li, et al., Chin. Chem. Lett. 36 (2025) 110191. doi: 10.1016/j.cclet.2024.110191
45. [45]
  X. Zhao, L. Ning, X. Zhou, et al., Anal. Chem. 93 (2021) 4894–4901. doi: 10.1021/acs.analchem.0c05081
46. [46]
  F. Yuan, A. Guo, L. Wang, et al., Angew. Chem. Int. Ed. (2025) e202501685.
47. [47]
  B. Zhu, X. Xing, J. Kim, et al., Biomaterials 304 (2024) 122419. doi: 10.1016/j.biomaterials.2023.122419
48. [48]
  M. Weng, X. Yang, Y. Ni, et al., Sens. Actuators B: Chem. 283 (2019) 769–775. doi: 10.1016/j.snb.2018.12.071
49. [49]
  S. Pang, J. Wu, F.F. Guo, et al., Anal. Chem. 96 (2024) 14800–14808. doi: 10.1021/acs.analchem.4c02364
50. [50]
  D. Shi, W. Liu, Y. Gao, et al., Nat. Aging 3 (2023) 297–312. doi: 10.1038/s43587-023-00360-x
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