A Naphthalimide-Based Fluorescent Probe for Detecting Intracellular pH and Its Biological Imaging Application

doi:10.6023/A20080399

Acta Chimica Sinica ›› 2021, Vol. 79 ›› Issue (1): 87-92.DOI: 10.6023/A20080399 Previous Articles Next Articles

Article

一种基于萘酰亚胺的检测细胞内pH值的荧光探针及其生物成像应用

任江波^a, 王蕾^b, 郭锐^a, 唐永和^a, 周红梅^a, 林伟英^a^,^c^,^*()

a 广西大学光功能材料与化学生物学研究院化学化工学院南宁 530004
b 南宁市食品药品检验所南宁 530007
c 济南大学光功能材料与成像研究院化学化工学院材料科学与工程学院济南 250022

投稿日期:2020-08-31 发布日期:2020-10-17
通讯作者: 林伟英
作者简介:
* E-mail: weiyinglin@163.com; Tel.: +86-0771-3236493; Fax: +86-0771-3236493
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)

A Naphthalimide-Based Fluorescent Probe for Detecting Intracellular pH and Its Biological Imaging Application

Jiangbo Ren^a, Lei Wang^b, Rui Guo^a, Yonghe Tang^a, Hongmei Zhou^a, Weiying Lin^a^,^c^,^*()

a Institute of Optical Materials and Chemical Biology, School of Chemistry and Chemical Engineering, Guangxi University, Nanning 530004, China
b Nanning Institute for Food and Drug Control, Nanning 530007, China
c Institute of Fluorescent Probes for Biological Imaging, School of Chemistry and Chemical Engineering, School of Materials Science and Engineering, Jinan University, Jinan 250022, China

Received:2020-08-31 Published:2020-10-17
Contact: Weiying Lin
Supported by:
the National Natural Science Foundation of China(21672083); the National Natural Science Foundation of China(21877048); the National Natural Science Foundation of China(22077048); the Startup Fund of Guangxi University(A3040051003)

1. A20080399.pdf(484KB)

Abstract

The pH of lysosome in cells has significant affect to various biological activities, including autophagy, phagocytosis, enzyme processing and so on. The metastasis and apoptosis of cancer cells are closely related to change of the lysosomal pH. The nucleus is the largest organelle in eukaryotic cells which control the most important genetic and metabolic processes, and the enzymes involved in metabolic processes are sensitive to changes of pH. Therefore, it is very important to investigate the pH change in cells. Herein, we designed and synthesized a novel fluorescent probe NpH-1 via a simple two-step reaction. The probe uses naphthalimide as the fluorescent group and morpholine group as the pH responsive site, and regulates the fluorescence through the photoinduced electron transfer (PET) mechanism. The spectroscopic properties of NpH-1 response to pH changes were measured in the Britton-Robison buffers, whose buffering range was from 1.81 to 11.92. In the pH range from 3.0 to 10.0, NpH-1exhibited a very rapid and reversible response to pH change. The pKa of NpH-1 was 5.41. It showed a very high photostability. Moreover, the fluorescence emission of NpH-1 was not influenced by biological species including metal ions, anions and amino acids. NpH-1 showed very low cytotoxicity, which indicated that it could be applied for cell imaging. Thus, we monitored the pH changes in HeLa cells via stimulating them by Chloroquine. The cell imaging results showed that NpH-1 could be utilized for detection of pH changes in living cells. Furthermore, the colocalization experiments of the probe NpH-1 for various organelles including lysosome, mitochondria, Golgi, endoplasmic reticulum and cellular nucleus were carried out, and the results exhibited the probe NpH-1 mainly distributed in lysosome and nucleus, which implied that the probe could be applied for detection of pH change in complex intracellular environment.

Key words: Lysosome, Cell nucleus, pH, Fluorescent imaging, Naphthalimide

Cite this article

Jiangbo Ren, Lei Wang, Rui Guo, Yonghe Tang, Hongmei Zhou, Weiying Lin. A Naphthalimide-Based Fluorescent Probe for Detecting Intracellular pH and Its Biological Imaging Application[J]. Acta Chimica Sinica, 2021, 79(1): 87-92.

Export EndNote|Reference Manager|ProCite|BibTeX|RefWorks

share this article

Fig. & Tab. 9

Scheme 1. The mechanism of the probe NpH-1 response to pH

Scheme 2. The synthesis of the probe NpH-1

Figure 1. The UV-Vis absorption spectra of the probeNpH-1 (10 μmol/L) in PBS buffers (pH range of 3.0~10.0).

Figure 2. (a) The emission spectra of the probeNpH-1 at 540 nm in pH range of 3.0~10.0; (b) the pH titration of the probeNpH-1. (λ ex=440 nm)

Figure 3. The curve of fluorescent emission intensity variation with time of the probeNpH-1(10 μmol/L)at pH 4.0, 7.4 and 9.0. (λ ex=440 nm, λ em=540 nm)

Figure 4. The reversible cycle of the fluorescent emission of the probe the probeNpH-1(10 μmol/L)between pH 4.0 and 8.0.

Figure 5. Fluorescence intensity ofNpH-1(10 μmol/L) atλ em=540 nm in solution at pH 4.0 and 7.4 with various species (200 mmol/L). 1.α-D-glucose; 2.L-phenylalanine; 3.L-alanine; 4.L-Methionine; 5. L-arginine; 6. L-ascorbic acid; 7.L-leucine; 8. L-proline; 9. L-tryptophan; 10.L-serine; 11.L-aspartic acid; 12.L-valine; 13.L-isoleucine; 14.L-histidine; 15. Methylglyoxal; 16. NaOAc; 17. CoCl2; 18. NaF; 19. glycine; 20. NaCNS; 21. Na2SO4; 22. KCl; 23. MnCl2; 24. NaCl; 25. Na2CO3; 26. NaHCO3; 27. MgCl2; 28. Na2S2O3•5H2O; 29. Cu2SO4•5H2O; 30. Na2SO3; 31. NaNO2.

Figure 6. (a~c) The imaging photograph of HeLa cell treated with the probeNpH-1 (10 μmol/L); (d~f) the imaging photograph of HeLa cell treated with chloroquine (10 μmol/L) for 30 min then addition of the probeNpH-1 (10 μmol/L).

Figure 7. (a1)~(a4) The colocalization photograph of the probe NpH-1 (10 μmol/L) with Hoechst 33342(10 μmol/L); (b1)~(b4) The colocalization photograph of the probeNpH-1(10 μmol/L) with MitoTrackerTM Deep Red (10 μmol/L); (c1)~(c4) The colocalization photograph of the probeNpH-1 (10 μmol/L) with LysoTrackerTM Deep Red; (d1)~(d4) The colocalization photograph of the probeNpH-1(10 μmol/L) with Golgi-Tracker Red; (e1)~(e4) The colocalization photograph of the probeNpH-1 (10 μmol/L) with ER-Tracker Blue.

References 34

[1]	Xu. Xu. K.; Klibanov A. M. J. Am. Chem. Soc. 1996, 118 (1), 9815.
[2]	Tian H.; Gan J.; Chen K.; He J.; Song Q.L.; Hou X. Y. J. Mater. Chem . 2002, 12 ( 5),1262.
[3]	Burleigh S. C.; van de Laar T.; Stroop C. J.; van Grunsven W. M.; O'Donoghue N.; Rudd P. M.; Davey G. P. BMC Biotechnol. 2011, 11(1), 95.
[4]	Zhou X.; Su F.; Lu H.; Senechal-Willis P.; Tian Y.; Johnson R. H.; Meldrum D. R. Biomaterials 2012, 33(1), 171.
[5]	Luzio J. P.; Pryor P. R.; Bright N. A. Nat. Rev. Mol. Cell Bio. 2007, 8 ( 8),622.
[6]	Reinheckel T.; Deussing J.; Roth W.; Peters C. Biol. Chem. 2001, 382 ( 5),735.
[7]	Watts C. Biochim. Biophys. Acta, Proteins Proteomics 2012, 1824(1), 14.
[8]	Zhao H. Traffic 2012, 13(10), 1307.
[9]	Mohamed M. M.; Sloane B. F. Nat. Rev. Cancer 2006, 6(10), 764.
[10]	Groth-Pedersen L.; Jaattela M. Cancer Lett. 2013, 332(2), 265.
[11]	Cirman T.; Oresic K.; Mazovec G. D.; Turk V.; Reed J. C.; Myers R. M.; Salvesen G. S.; Turk B. J. Biol. Chem. 2004, 279 ( 5),3578.
[12]	Hesse S. J.A.; Ruijter G. J.G.; Dijkema C.; Visser J. J. Biotechnol. 2000, 77(1), 5.
[13]	Schartner E. P.; Henderson M. R.; Purdey M.; Dhatrak D.; Monro T. M.; Gill P. G.; Callen D. F. Cancer Res. 2016, 76 ( 23),6795.
[14]	Newport J. W.; Forbes D. J. Annu. Rev. Biochem. 1987, 56(1), 535.
[15]	Shen W. H.; Balajee A. S.; Wang J.; Wu H.; Eng C.; Pandolfi P. P.; Yin Y. Cell 2007, 128(1), 157.
[16]	Zhou C. M.S. Thesis, Liaoning University, Shen-yang , 2013.
	(周超, 硕士论文, 辽宁大学, 沈阳,2013). (in Chinese)
[17]	Singha S.; Kim D.; Seo H.; Cho S.W.; Ahn K. H. Chem. Soc. Rev. 2015, 44 ( 13),4367.
[18]	Li X.; Gao X.; Shi W.; Ma H. Chem. Rev. 2014, 114(1), 590.
[19]	Lv H.; Yang X.F.; Zhong Y.; Guo Y.; Li Z.; Li H. Anal. Chem. 2014, 86 ( 3),1800.
[20]	Carter K. P.; Young A. M.; Palmer A. E. Chem. Rev. 2014, 114 ( 8),4564.
[21]	Li L.; Li P.; Fang J.; Li Q.; Xiao H.; Zhou H.; Tang B. Anal. Chem. 2015, 87(12), 6057.
[22]	Sun R.; Liu W.; Xu Y.; Lu J.; Ge J.; Ihara M. Chem. Commun. 2013, 49(91), 10709.
[23]	Zhang Y.; Fang H.; Zhang X.; Wang S.; Xing G. ChemistrySelect 2016, 1(1), 1.
[24]	Wang, X.L.; Li, X.J.; Sun, R.; Xu, Y.J.; Ge J. F. Analyst. 2016, 141(10), 2962.
[25]	Song G. J.; Bai S.Y.; Luo J.; Cao X. Q.; Zhao B. X. J. Fluoresc . 2016, 26 ( 6),2079.
[26]	Zhang S.; Chen T.; Lee H.; Bi J.; Ghosh A.; Fang M.; Qian Z.; Xie F.; Ainsley J.; Christov C.; Luo F.; Zhao F.; Liu H. ACS Sensors 2017, 2(7), 924.
[27]	Zhang, Y; Wang, Z.-L; Tao,Y; Xu-X; Fang-H; Wang,S.-F. Chin. J. Org. Chem. 2018, 38(10), 2693 . (in Chinese)
	张燕, 王忠龙, 陶钰, 徐徐, 方华, 王石发, 有机化学, 2018, 38(10), 2693.
[28]	Xiang, D.-C; Liu, H; Meng, Q.-H; Lan, M.-B; Wei, G. Acta Chim. Sinica 2013, 71(10), 1435 . (in Chinese)
	向德成, 刘恒, 孟庆华, 蓝闽波, 卫钢, 化学学报, 2013, 71(10), 1435.
[29]	Wu, P.-Q; Liang, C.-H. Int. J. Med. Radiol. 2016, 39(04), 410 . (in Chinese)
	吴佩琪, 梁长虹. 国际医学放射学杂志, 2016, 39(04), 410.
[30]	Guan, X.-L; Li, Z.-F; Wang, L; Liu, M.-N; Wang, K.-L; Yang, X.-Q; Li, Y.-L; Hu, L.-L; Zhao, X.-L; Lai, S.-J; Lei, Z.-Q. Acta Chim. Sinica 2019, 77(12), 1268 . (in Chinese)
	关晓琳, 李志飞, 王林, 刘美娜, 王凯龙, 杨学琴, 李亚丽, 胡丽丽, 赵小龙, 来守军, 雷自强, 化学学报, 2019, 77(12), 1268.
[31]	Ma, Y; Chen, K.-X; Guo, Z.-L; Liu, S.-J; Zhao, Q; Huang, W.-Y. Acta Chim. Sinica 2020, 78(01), 23 . (in Chinese)
	马云, 陈可欣, 郭则灵, 刘淑娟, 赵强, 黄维扬, 化学学报, 2020, 78(01), 23.
[32]	Li, Y; Zhan, H; Li, M. New Chem. Mater. 2019, 47(08), 135 . (in Chinese)
	李阳, 张辉, 李明, 化工新型材料, 2019, 47(08), 135.
[33]	Liu Y.; Zhou J.; Wang L.; Hu X.; Liu X.; Liu M.; Cao Z.; Shangguan D.; Tan W. J. Am. Chem. Soc. 2016, 138 ( 38),12368.
[34]	Cao, X.-J; Chen, L.-N; Zhang, X; Liu, J.-T; Chen, M.-Y; Wu, Q.-R; Miao, J.-Y; Zhao, B.-X. Anal. Chim. Acta 2016, 920, 86.

一种基于萘酰亚胺的检测细胞内pH值的荧光探针及其生物成像应用

A Naphthalimide-Based Fluorescent Probe for Detecting Intracellular pH and Its Biological Imaging Application

RichHTML

PDF

Supporting Info.

Knowledge

Abstract

Cite this article

share this article

Fig. & Tab. 9

References 34

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	Jianqiang Chen, Gangguo Zhu, Jie Wu. Recent Advances in Nickel-Catalyzed Ring Opening Cross-Coupling of Aziridines [J]. Acta Chimica Sinica, 2024, 82(2): 190-212.
[2]	Cheng-Qiang Wang, Chao Feng. Applications of Nucleophilic Fluorine Sources in the Selective Fluorofunctionalization of Unsaturated Carbon-Carbon Bonds [J]. Acta Chimica Sinica, 2024, 82(2): 160-170.
[3]	Jinglin Yi, Mao Chen. Photo-Induced Copolymerization of Chlorotrifluoroethylene and Methyl Isopropenyl Ether^★ [J]. Acta Chimica Sinica, 2024, 82(2): 126-131.
[4]	Zhiqiang Wang, Jinzhan Su. Investigation of the Kinetic Properties and Photoelectrochemical Water Splitting of Cu₃V₂O₈/ZnO Photoanode Modified by Cobalt Phosphate [J]. Acta Chimica Sinica, 2024, 82(1): 26-35.
[5]	Chenyu Liao, Shanwei Guo, Meiwei Huang, Yong Guo, Qing-Yun Chen, Chao Liu, Yunwen Zhang. Synthesis and Properties of Fluoroether Phosphocholine [J]. Acta Chimica Sinica, 2024, 82(1): 46-52.
[6]	Yuqing Shi, Mingzhu Chu, Bo Han, Haojie Ma, Ran Li, Xueyan Hou, Yuqi Zhang, Ji-Jiang Wang. Smart Two-dimensional Photonic Crystal Hydrogel for Accurate Detection of Hg²⁺ [J]. Acta Chimica Sinica, 2024, 82(1): 9-15.
[7]	Ruxin Zeng, Peng R. Chen. RNA-Binding Proteome Analysis and Functional Explorations^★ [J]. Acta Chimica Sinica, 2024, 82(1): 53-61.
[8]	Ye Tian, Duanhui Si, Shuiying Gao, Rong Cao. Ultra-Long Organic Room Temperature Phosphorescence of Phthalic Acid Derivative Modified Polymer^★ [J]. Acta Chimica Sinica, 2023, 81(9): 1129-1134.
[9]	Yuhan Wu, Dongdong Zhang, Hongyu Yin, Zhengnan Chen, Wen Zhao, Yuhua Chi. Density Functional Theory Study of Janus In₂S₂X Photocatalytic Reduction of CO₂ under “Double Carbon” Target [J]. Acta Chimica Sinica, 2023, 81(9): 1148-1156.
[10]	Fengjie Ge, Kaizhi Zhang, Qingpeng Cao, Hui Xu, Tao Zhou, Wenhao Zhang, Xinxin Ban, Xiaobo Zhang, Na Li, Peng Zhu. Design, Synthesis and Electroluminescence Performance of Flexible Fluorenyl Block Delayed Fluorescence Dimers [J]. Acta Chimica Sinica, 2023, 81(9): 1157-1166.
[11]	Jiao Kong, Lin Du, Xiangyang Li, Jidong Zhu, Ya-Qiu Long. Small Molecule Degraders Targeting the SHP2^E76A Mutant Effectively Inhibiting the Proliferation of Wild-type and Mutant SHP2 Dependent Tumor Cells [J]. Acta Chimica Sinica, 2023, 81(9): 1120-1128.
[12]	Yuchun Han, Yilin Wang. Retrospect and Prospect of Long-lasting Antibacterial Materials^★ [J]. Acta Chimica Sinica, 2023, 81(9): 1196-1201.
[13]	Ruxin Tian, Miao Yang, Guo Chen, Jiangshan Liu, Mengmei Yuan, Hong Yuan, Shuxin Ouyang, Tierui Zhang. Ru/Quartz Filter Paper: A Recyclable Photothermocatalytic Film for CO₂ Methanation^★ [J]. Acta Chimica Sinica, 2023, 81(8): 869-873.
[14]	Mei Hong, Jinqiang Gao, Tong Li, Shihe Yang. In-situ Etching Strategy for Manipulation of Hierarchical Zeolite and Its Application^★ [J]. Acta Chimica Sinica, 2023, 81(8): 937-948.
[15]	Yongxue Li, Yu Liu. Supramolecular Secondary Assembly Based on Amphiphilic Calix[4]arenes and Its Biological Applications^★ [J]. Acta Chimica Sinica, 2023, 81(8): 928-936.