2. Academic Validation
  3. A facile fluorescence-coupling approach to visualizing leonurine uptake and distribution in living cells and Caenorhabditis elegans

A facile fluorescence-coupling approach to visualizing leonurine uptake and distribution in living cells and Caenorhabditis elegans

  • Phytomedicine. 2024 May 14:130:155737. doi: 10.1016/j.phymed.2024.155737.
Hao Shi 1,  Jinrong Yang 2,  Jiajie Lin 1,  Xiaobing Hong 3,  Ziyuan Zhou 4,  Jiamin Zhao 1,  Yiwen Li 1,  Junjie Li 1,  Chaofeng Wu 1,  Jinwu Yan 5,  Nai-Kei Wong 6,  Lei Gao 7
Affiliations

Affiliations

  • 1 School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, Guangdong 510515, China.
  • 2 School of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, China.
  • 3 Department of Pharmacy, The Second Affiliated Hospital, Shantou University Medical College, Shantou, China.
  • 4 National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital & Shenzhen Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Shenzhen 518116, China.
  • 5 School of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, China. Electronic address: yjw@scut.edu.cn.
  • 6 Clinical Pharmacology Section, Department of Pharmacology, Shantou University Medical College, Shantou, Guangdong 515041, China. Electronic address: wongnk@stu.edu.cn.
  • 7 School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, Guangdong 510515, China; Guangdong Provincial Key Laboratory of Chinese Medicine Pharmaceutics, Southern Medical University, Guangzhou, Guangdong 510515, China; Guangdong Basic Research Center of Excellence for Integrated Traditional and Western Medicine for Qingzhi Diseases, Guangzhou, Guangdong 510515, China. Electronic address: raygaolei@smu.edu.cn.
PMID: 38772183 DOI: 10.1016/j.phymed.2024.155737
Abstract

Background: Caenorhabditis elegans (C. elegans) has been recognized for being a useful model organism in small-molecule drug screens and drug efficacy investigation. However, there remain bottlenecks in evaluating such processes as drug uptake and distribution due to a lack of appropriate chemical tools.

Purpose: This study aims to prepare fluorescence-labeled leonurine as an example to monitor drug uptake and distribution of small molecule in C. elegans and living cells.

Methods: FITC-conjugated leonurine (leonurine-P) was synthesized and characterized by LC/MS, NMR, UV absorption and fluorescence intensity. Leonurine-P was used to stain C. elegans and various mammalian cell lines. Different concentrations of leonurine were tested in conjunction with a competing parent molecule to determine whether leonurine-P and leonurine shared the same biological targets. Drug distribution was analyzed by imaging. Fluorometry in microplates and flow cytometry were performed for quantitative measurements of drug uptake.

Results: The UV absorption peak of leonurine-P was 490∼495 nm and emission peak was 520 nm. Leonurine-P specifically bound to endogenous protein targets in C. elegans and mammalian cells, which was competitively blocked by leonurine. The highest enrichment levels of leonurine-P were observed around 72 h following exposure in C. elegans. Leonurine-P can be used in a variety of cells to observe drug distribution dynamics. Flow cytometry of stained cells can be facilely carried out to quantitatively detect probe signals.

Conclusions: The strategy of fluorescein-labeled drugs reported herein allows quantification of drug enrichment and visualization of drug distribution, thus illustrates a convenient approach to study phytodrugs in pharmacological contexts.

Keywords

Caenorhabditis elegans; Drug uptake; Fluorescence labelling; Leonurine; Plant-derived natural product.

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