Liver-Targeted Fluorescent Probes for Specific Detection of ONOO- in HepG2 Cells

doi:10.19894/j.issn.1000-0518.220268

Chinese Journal of Applied Chemistry ›› 2023, Vol. 40 ›› Issue (3): 441-448.DOI: 10.19894/j.issn.1000-0518.220268

• Full Papers • Previous Articles Next Articles

Liver-Targeted Fluorescent Probes for Specific Detection of ONOO^- in HepG2 Cells

Jia-Mei GENG¹^,², Su-Fang MA¹, Wen LIU¹, Hai-Peng DIAO¹, Zhi-Fang WU², Si-Jin LI²()

^1.School of Basic Medical Science，Shanxi Medical University，Taiyuan 030001，China
^2.Department of Nuclear Medicine，First Hospital of Shanxi Medical University，Taiyuan 030001，China

Received:2022-08-08 Accepted:2022-11-21 Published:2023-03-01 Online:2023-03-27
Contact: Si-Jin LI
About author:lisjnm123@163.com
Supported by:
the National Natural Science Foundation of China(82202289);the Natural Science Foundation of Shanxi Province(201901D111191)

1. .pdf(811KB)

Abstract

Abstract:

As an important biologically active molecule， peroxynitrite （ONOO^-） plays an important role in cellular signal transduction and homeostatic regulation. Precise and targeted detection of ONOO^- in biological systems is crucial for in-depth exploration of its physiological and pathological mechanisms. A “turn-on” fluorescent probe naphthalamide derivative （NA） is designed based on 1，?8-naphthalene imide and ursodeoxycholic acid for specific detection of ONOO^- in HepG2 cells. Probe NA is synthesized by three steps， the structures of intermediates and target compounds are confirmed by ¹H NMR and ¹³C NMR spectroscopy and HRMS. The optical properties of probe NA are explored. The green fluorescence of probe NA in the presence of ONOO^- is significantly increased compared to its fluorescence in the absence of ONOO^-. The fluorescence intensity and concentrations of ONOO^- shows a good linear relationship in the range of 0.0 to 14.0 μmol/L， with the detection limit of 0.27 μmol/L. In addition， probe NA shows excellent selectivity toward ONOO^-， good biocompatibility， and the fluorescence intensity is almost independent to the pH of the environment. Finally， probe NA is applied to monitor ONOO^- in living cells. The results of cell fluorescence imaging exhibit that probe NA possesses good targeting ability to HepG2 cells， and can detect exogenous and endogenous ONOO^- in HepG2 cells， which demonstrates the great potential of in the application of ONOO^- detection in vivo.

Key words: Peroxynitrite, Liver targeting, Fluorescent probes, Fluorescence imaging

CLC Number:

O625

Jia-Mei GENG, Su-Fang MA, Wen LIU, Hai-Peng DIAO, Zhi-Fang WU, Si-Jin LI. Liver-Targeted Fluorescent Probes for Specific Detection of ONOO^- in HepG2 Cells[J]. Chinese Journal of Applied Chemistry, 2023, 40(3): 441-448.

Figures/Tables 8

Fig.1 Synthetic route of probe NA

Fig.2 Fluorescence emission spectra of probe NA （10 μmol/L） before （a） and after （b） responding to ONOO-（100 μmol/L）

Fig.3 （A） Fluorescence emission spectra of probe NA （10 μmol/L） reacting with different concentrations of ONOO-（λex=451 nm）；（B） Linearity curve between the fluorescence intensity of probe NA and the concentration of ONOO-

Fig.4 Fluorescence intensity of probe NA（10 μmol/L） mixing with different reactive substances （λex=451 nm）

Fig.5 Effect of pH on probe NA （10 μmol/L） before （a） and after （b） responding to ONOO- （100 μmol/L）（λex=451 nm）

Fig.6 The viability of HepG2 cells incubated with different concentration of probe NA

Fig.7 Fluorescence imaging of （A） A549 cells，（B） HCT116 cells and （C） HepG2 cells incubated with the probe NA （30 μmol/L） for 30 min； D-F： merged images of A-C； λex=488 nm， λem=500~600 nm

Fig. 8 Fluorescence imaging of HepG2 cells：（A） incubated with the probe NA （30 μmol/L） for 30 min；（B） Incubated with SIN （800 μmol/L） for 30 min， and then incubated with the probe NA （30 μmol/L） for another 30 min；（C） Incubated with LPS （20 μg/mL） and IFN-γ （300 ng/mL） for 12 h， and then incubated with the probe NA （30 μmol/L） for another 30 min；（D） Incubated with TEMPO （1080 μmol/L）， LPS （20 μg/mL） and IFN-γ （300 ng/mL） for 12 h， and then incubated with the probe NA （30 μmol/L） for another 30 min；（E-H merged images of A-D； λex=488 nm， λex=500~600 nm；（I） The relative fluorescence intensity ratio under different culture conditions

References 28

1	ZHOU W， YANG W， FAN K， et al. A hypoxia-activated NO donor for the treatment of myocardial hypoxia injury［J］. Chem Sci， 2022， 13（12）： 3549-3555.
2	DU Z， ZHANG X， GUO Z， et al. X-ray-controlled generation of peroxynitrite based on nanosized LiLuF₄∶Ce³⁺ scintillators and their applications for radiosensitization［J］. Adv Mater， 2018， 30（43）： 1804046.
3	SUN Q， XU J， JI C， et al. Ultrafast detection of peroxynitrite in Parkinson′s disease models using a near-infrared fluorescent probe［J］. Anal Chem， 2020， 92（5）： 4038-4045.
4	FENG J， CHEN X， GUAN B， et al. Inhibition of peroxynitrite-induced mitophagy activation attenuates cerebral ischemia-reperfusion injury［J］. Mol Neurobiol， 2018， 55（8）： 6369-6386.
5	WANG Z， WANG W， WANG P， et al. Highly sensitive near-infrared imaging of peroxynitrite fluxes in inflammation progress［J］. Anal Chem， 2021， 93（5）： 3035-3041.
6	ZENG X， LI Z， FU J， et al. A novel ultrasensitive peroxynitrite-specific fluorescent probe and its bioimaging applications in living systems［J］. Dyes Pigm， 2021， 186： 108982.
7	张成路，王一鸣，任芷漩，等. 以苯并咪唑萘酰亚胺为荧光团高选择性快速检测H₂S的荧光探针［J］.应用化学，2022， 39（3）： 489-497.
	ZHANG C L， WANG Y M， REN Z X， et al. Fluorescent probe for rapid detection of H₂S with benzimidazole naphthalimide as the core［J］. Chin J Appl Chem， 2022， 39（3）： 489-497.
8	DOU W T， HAN H H， SEDGWICK A C， et al. Fluorescent probes for the detection of disease-associated biomarkers［J］. Sci Bull， 2022， 67（8）： 853-878.
9	WANG J， ZHANG J， WANG J， et al. Fluorescent peptide probes for organophosphorus pesticides detection［J］. J Hazard Mater， 2020， 389： 122074.
10	XU W， YANG Q， ZENG J， et al. A biomarker （ONOO^-）-activated multicolor fluorescent probe for early detection and assessment of arthritis［J］. Sens Actuators B： Chem， 2022， 359： 131565.
11	SUN Y， TANG X， LI X， et al. PET-ESIPT-based fluorescent probes for revealing the fluctuation of peroxynitrite （ONOO^-） in living cells， zebrafishes and brain tissues［J］. Sens Actuators B： Chem， 2022， 353： 131121.
12	GU J， LIU Y， SHEN J W， et al. A three-channel fluorescent probe for selective detection of ONOO^- and its application to cell imaging［J］. Talanta， 2022， 244： 123401.
13	XIA Q， FENG S， HONG J， et al. One probe for multiple targets： a NIR fluorescent rhodamine-based probe for ONOO^- and lysosomal pH detection in live cells［J］. Sens Actuators B： Chem， 2021， 337： 129732.
14	WU L， LIU J， TIAN X， et al. Dual-channel fluorescent probe for the simultaneous monitoring of peroxynitrite and adenosine-5′-triphosphate in cellular applications［J］. J Am Chem Soc， 2022， 144 （1）： 174-183.
15	DENG Y， FENG G. Visualization of ONOO^- and viscosity in drug-induced hepatotoxicity with different fluorescence signals by a sensitive fluorescent probe［J］. Anal Chem， 2020， 92（21）： 14667-14675.
16	JIANG W L， LI Y， WANG W X， et al. A hepatocyte-targeting near-infrared ratiometric fluorescent probe for monitoring peroxynitrite during drug-induced hepatotoxicity and its remediation［J］. Chem Commun， 2019， 55 （95）： 14307-14310.
17	LIU C， DUAN Q， ZHANG X， et al. A novel hepatoma-specific fluorescent probe for imaging endogenous peroxynitrite in live HepG2 cells［J］. Sens Actuators B： Chem， 2019， 289： 124-130.
18	YANG Y X， ZHANG L， LI J J， et al. In vivo imaging via a red-emitting fluorescent probe to diagnosing liver cancer or drug-induced liver disease［J］. Anal Chim Acta， 2021， 1168： 338621.
19	CHEN J， JIANG Z， ZHANG Y S， et al. Smart transformable nanoparticles for enhanced tumor theranostics［J］. Appl Phys Rev， 2021， 8（4）： 041321.
20	杨佳臻，邹昊洋，丁建勋，等. 胱氨酸基聚氨基酸纳米材料的可控合成及其生物医学应用［J］. 高分子学报， 2021， 52（8）： 960-977.
	YANG J Z， ZOU H Y， DING J X， et al. Controlled synthesis and biomedical applications of cystine-based polypeptide nanomaterials［J］. Acta Polym Sin， 2021， 52（8）： 960-977.
21	杨佳臻，丁建勋. 抗肿瘤纳米材料［J］. 应用化学， 2022， 39（5）： 855-856.
	YANG J Z， DING J X. Anti-tumor nanomaterials［J］. Chin J Appl Chem， 2022， 39（5）： 855-856.
22	LI Z， LU J， PANG Q， et al. Construction of a near-infrared fluorescent probe for ratiometric imaging of peroxynitrite during tumor progression［J］. Analyst， 2021， 146： 5204-5211.
23	XIA L， TONG Y， LI L， et al. A selective fluorescent turn-on probe for imaging peroxynitrite in living cells and drug-damaged liver tissues［J］. Talanta， 2019， 204： 431-437.
24	DENG Y， WANG Y， JIA F， et al. Tailoring supramolecular prodrug nanoassemblies for reactive nitrogen species-potentiated chemotherapy of liver cancer［J］. ACS Nano， 2021， 15（5）： 8663-8675.
25	ZHAO M， LI B， WU Y， et al. A tumor-microenvironment-responsive lanthanide-cyanine FRET sensor for NIR-II luminescence-lifetime in situ imaging of hepatocellular carcinoma［J］. Adv Mater， 2020， 2001172.
26	XIAO L X， YU E D， YUE H L， et al. Enhanced liver targeting of camptothecin via conjugation with deoxycholic acid［J］. Molecules， 2019， 24 （6）： 1179.
27	ZHANG Z， CAI H X， LIU Z J， et al. Effective enhancement of hypoglycemic effect of insulin by liver-targeted nanoparticles containing cholic acid-modified chitosan derivative［J］. Mol Pharmaceutics， 2016， 13（7）： 2433-2442.
28	GONG T， CHENG R N， WANG X Y， et al. Supramolecular-interaction-mediated aggregation of anticarcinogens on triformyl cholic acid-functionalized Fe₃O₄ nanoparticles and their dual-targeting treatment for liver cancer［J］. New J Chem， 2021， 45（15）： 6880-6888.

[1]	Song-Tao ZHANG, Ying-Hui WANG, Hong-Jie ZHANG. Nd³⁺ Sensitized Fluorescent Nanoprobes for Vascular Imaging in the Second Near Infrared Window [J]. Chinese Journal of Applied Chemistry, 2022, 39(4): 685-693.
[2]	Si-Wei YU, Liang-Peng WANG, Ri-Zhe JIN, Chuan-Qing KANG. Recognition of ClO^－ and Cellular Imaging with Xanthene-based Fluorescent Probes [J]. Chinese Journal of Applied Chemistry, 2022, 39(12): 1903-1911.
[3]	YAO Qianfang,CHENG Wenyu,YIN Meizhen. Host-Guest Supramolecular Fluorescent Probes Based on Macrocyclic Molecules [J]. Chinese Journal of Applied Chemistry, 2017, 34(12): 1344-1354.
[4]	BU Lulu,WANG Qing,XIE Yongshu. Research Progress of Fluorescent Zinc Probes [J]. Chinese Journal of Applied Chemistry, 2017, 34(12): 1355-1369.
[5]	CHEN Jianhua,LI Dongyang,LIU Shenghua,TAN Ying,YIN Jun. Recent Advances in Phosgene and Nerve Agents Responsive Fluorescent Probes [J]. Chinese Journal of Applied Chemistry, 2017, 34(12): 1413-1432.
[6]	Jia ZHOU, Yun NI, Chengwu ZHANG, Xinghan QIU, Yanfei ZHAO, Lei BAI, Gaobin ZHANG, Lin LI. Design and Synthesis of Pyrimidine Based Two-Photon Fluorescence Probe and Its Application in Bioimaging [J]. Chinese Journal of Applied Chemistry, 2017, 34(12): 1450-1456.
[7]	Jia ZHOU, Yun NI, Chengwu ZHANG, Xinghan QIU, Yanfei ZHAO, Lei BAI, Gaobin ZHANG, Lin LI. Design and Synthesis of Pyrimidine Based Two-Photon Fluorescence Probe and Its Application in Bioimaging [J]. Chinese Journal of Applied Chemistry, 2017, 34(12): 0-0.
[8]	LI Haidong,YAO Qichao,FAN Jiangli,DU Jianjun,PENG Xiaojun. Research Advance on the Fluorescent Probe of Palladium Ions [J]. Chinese Journal of Applied Chemistry, 2016, 33(10): 1099-1114.

Liver-Targeted Fluorescent Probes for Specific Detection of ONOO^- in HepG2 Cells

RichHTML

PDF

Supplement

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 8

References 28

Related Articles 8

Recommended Articles

Metrics

Comments