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Recent advancements in single-cell metabolic analysis for pharmacological research

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Journal of Pharmaceutical Analysis 2023年第10期13卷 1102-1116页

作者： Ying Hou Hongren Yao jin-ming lin Beijing Key Laboratory of Microanalytical Methods and Instrumentation Key Laboratory of Bioorganic Phosphorus Chemistry&Chemical Biology(Ministry of Education)Department of ChemistryTsinghua UniversityBeijing100084China

Cellular heterogeneity is crucial for understanding tissue biology and disease *** research is being advanced by single-cell metabolic analysis,which offers a technique to identify variations in RNA,proteins,metabolites,and drug molecules in *** this review,the recent advancement of single-cell metabolic analysis techniques and their applications in drug metabolism and drug response are ***-precision and controlled single-cell isolation and manipulation are provided by microfluidics-based methods,such as droplet microfluidics,microchamber,open microfluidic probe,and digital *** are used in tandem with variety of detection techniques,including optical imaging,Raman spectroscopy,electrochemical detection,RNA sequencing,and mass spectrometry,to evaluate single-cell metabolic changes in response to drug *** advantages and disadvantages of different techniques are discussed along with the challenges and future directions for single-cell *** techniques are employed in pharmaceutical analysis for studying drug response and resistance pathway,therapeutic targets discovery,and in vitro disease model evaluation.

关键词： Single-cell Metabolism Pharmacology Drug response Microfluidics

Self-catalytic degradation of ortho-chlorophenol with Fenton's reagent studied by chemiluminescence

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Journal of Environmental Sciences 2012年第3期24卷 550-557页

作者： Zhen lin Hui Chen Yun Zhou Nobuaki Ogawa jin-ming lin Beijing Key Laboratory of Microanalytical Methods and Instrumentation Department of ChemistryTsinghua UniversityBeijing 100084China. Faculty of Engineering and Resource Science Akita UniversityTegata GakuenchoAkita010-8502Japan

The degradation of ortho-chlorophenol using Fenton＇s reagent was studied by chemiluminescence （CL）. Without a special CL reagent, a weak CL emission from the mixture of ferrous ion and hydrogen peroxide was observed at room temperature. The CL intensity was increased by the addition of ortho-chlorophenol into the mixed solution. When the temperature was raised to 65°C, the CL intensity was enhanced strongly. The CL mechanisms for the system H2O2-Fe2＋ with and without ortho-chlorophenol were studied by examining the CL spectrum, gas chromatography-mass spectrometry and electron spin resonance spectrum. The effects of various free radical scavengers, surfactants and fluorescence compounds on the CL intensity were also investigated. A self-catalytic oxidation mechanism was proposed. The results showed that singlet oxygen was the main emitter for the system H2O2-Fe2＋. The strong CL from the system H2O2-Fe2＋-ortho-chlorophenol was due to singlet oxygen and electronically excited quinone. The benzenediol-like intermediate product formed during the phenol oxidation process greatly promoted the Fenton＇s reaction and led to higher CL intensity. Chemiluninescence is a novel approach for the investigation of the oxidation of some organic pollutants by Fenton＇s reagent.

关键词： ortho-chlorophenol Fenton’ reagent chemiluminescence singlet oxygen

Inhibition of anaerobic probiotics on colorectal cancer cells using intestinal microfluidic systems

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Science China Chemistry 2018年第8期61卷 1034-1042页

作者： lin Zhou Sifeng Mao Qiushi Huang Xiangwei He jin-ming lin College of Biological Sciences and Biotechnology Beijing Forestry University Beijing Key Laboratory of Microanalytical Methods and Instrumentation Department of Chemistry Tsinghua University

Intestinal flora play an important role in human's immune system. Many bacteria adhere to the wall of the testinal wall. These Intestinal flora help digestion, and also stop their disease-causing counterparts from invading. Most of the current researches focused on the interaction between cells and the construction of organs, but few researches studied on the role of microorganisms and cells. Here, we developed an in vitro living cell systems to simulate the structure, absorption, transport and pathophysiological characteristics of the human intestinal tract and the key microbial symbiosis. The co-culture of Clostridium butyricum(***) and colon cancer cells showed a different immune effect. C. butyricum could inhibit the proliferation of HCT116 cells, cause cell cycle arrest and promote apoptosis. But it had no significant effect on Caco-2 cells. Thus, basic functional characteristics of the gut were successfully simulated in a controlled microfluidic system. This approach is suggested as a powerful method in the investigation on drug metabolism and intestinal diseases.

关键词： microfluidics intestinal microorganisms co-culture metabolism

Imitation of drug metabolism in cell co-culture microcapsule model using a microfluidic chip platform coupled to mass spectrometry

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Chinese Chemical Letters 2020年第2期31卷 451-454页

作者： Yajing Zheng Zengnan Wu jin-ming lin ling lin Beijing Key Laboratory of Microanalytical Methods and Instrumentation Department of ChemistryTsinghua UniversityBeijing 100084China State Key Laboratory of Chemical Resource Engineering Beijing University of Chemical TechnologyBeijing 100029China CAS Key Laboratory of Standardization and Measurement for Nanotechnology CAS Center for Excellence in NanoscienceNational Center for Nanoscience and TechnologyBeijing 100190China

In this work,a multi-functional analysis platform by coupling a microfluidic chip to a mass spectrometry(MS) detector was *** constructed a three-dimensional tumor-endothelial co-culture model for simulating drug resistance during tumor *** this specially designed integrated platform,the first step was to prepare heterogeneous cell-encapsulated alginate microcapsules for threedimensional co-culture,and the second step was to achieve on-line perfusion culture and continuous drug stimulation on *** facilitates cell proliferation analysis and the collection of metabolism *** micro solid phase extraction column(SPE) pretreatment,subsequent mass spectrometry could detect drug *** high activity of two kinds of cells(A549 and HUVEC) shows the biocompatibility of the *** was used as a model drug,the distinctions of drug absorption between the mono-culture group and co-culture group were clearly observed by electrospray ionization quadrupole time-of-flight mass spectrometry(ESI-Q-TOF MS).Therefore,the integrated platform has shown promise as a high throughput,low cost for cell metabolism research and drug screening processes.

关键词： Microfluidic chip ESI-Q-TOF MS Co-culture Drug metabolism Droplet technique Hydrogel microcapsule

Di-4-ANEPPDHQ probes the response of lipid packing to the membrane tension change in living cells

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Chinese Chemical Letters 2022年第3期33卷 1377-1380页

作者： Nan Li Weifei Zhang Haifeng lin jin-ming lin Beijing Key Laboratory of Microanalytical Methods and Instrumentation MOE Key Laboratory of Bioorganic Phosphorus Chemistry&Chemical BiologyDepartment of ChemistryTsinghua UniversityBeijing 100084China Division of Chemical Metrology and Analytical Science National Institute of MetrologyBeijing 100029China

Membrane tension plays a significant role in many cellular processes including cell adhesion, migration and spreading. Despite the importance of membrane tension, it remains difficult to measure in vivo. Recently, the development of non-invasive fluorescent probes have made great progress, especially excitedstate deplanarization in molecular rotors has been applied to image membrane tension in living ***, an intrinsic limitation of such kind of probe is that they depend on the lipid packing, and how the lipid packing responds to the membrane tension change remains unclear. Therefore, in this work,we used a polarity-sensitive membrane probe to investigate the possible response mechanism of lipid packing to the change of membrane tension that was regulated by osmotic shocks. The results showed that an increase in membrane tension could stretch the lipids apart with large displacements, and this change was not homogeneous on the whole membrane, instead, increase of membrane tension induced phase separation.

关键词： Lipid packing Membrane tension Osmotic shocks Polarity-sensitive probe Phase separation

Development of LC–MS method for analysis of paclitaxel-inhibited growth and enhanced therapeutic response in human glioblastoma cells

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Chinese Chemical Letters 2015年第10期26卷 1225-1230页

作者： Cai-Hou lin Xue-Xia lin ling lin Jun-ming Wang Zhi-Xiong lin jin-ming lin Department of Neurosurgery The First Affiliated Hospital of Fujian Medical University Beijing Key Laboratory of Microanalytical Methods and Instrumentation Department of Chemistry Tsinghua University Department of Bioengineering School of Engineering The University of Tokyo

Glioma stem cells are considered responsible for drug resistance and glioma relapse resulting in poor prognosis in glioblastoma multiforme. SU3 glioma cell is a highly invasive glioma stem cell line from the patients with glioblastoma multifrome. It is of great significance to study the efficacy and molecular mechanism for anticancer drug effects on SU3 glioma cells. In this work, we develop a liquid chromatography–mass spectrometry（LC–MS） method for direct analysis of the role of drugs（paclitaxel）on SU3 glioma cells at the molecular level. We use the specific fluorescence dyes to evaluate cell viability,the levels of ROS and GSH when the cells were treated with drugs. In addition, the LC–MS platform was successfully employed to detect the amount of 6-O-methylguanine, demonstrating that it is effective to induce cell apoptosis and enhance the cytotoxic response of SU3 glioma cells. The analytical linear equals are Y = 9.49 ？ 105 X ＋ 2.42 ？ 104 for 6-O-methylguanine（R2= 0.9998） and Y = 4.72 ？ 104 X ＋ 2.21 ？ 103（R2= 0.9996） for 7-methylguanine. Thus, the combination of cell-specific fluorescence dyes and LC–MS method enables us to reveal the molecular mechanism of paclitaxel-inhibited growth and enhanced therapeutic response in the chemotherapy for glioma multiforme.

关键词： LC–MS Drug analysis SU3 glioma cell Paclitaxel Cell cycle arrest ROS assay

An on-chip intestine-liver model for multiple drugs absorption and metabolism behavior simulation

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Science China Chemistry 2018年第2期61卷 236-242页

作者： mingsha Jie Haifeng lin Ziyi He Hanyang Liu Haifang Li jin-ming lin State Key Laboratory of Chemical Resource Engineering Beijing University of Chemical Technology Department of Chemistry Beijing Key Laboratory of Microanalytical Methods and Instrumentation Key Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology Ministry of Education Tsinghua University

A double-layer microfluidic chip integrated with a hollow fiber(HF)was developed to reconstitute the intestine-liver functionality for studying the absorption and metabolism of combination ***-2 cells were inoculated in the HF cavity at the top of the serpentine channel to simulate the intestinal tissue for drug absorption and transport studied,and Hep G2 cells,seeded in the bottom chamber,were used to mimic the liver for metabolism-related *** and dacarbazine were selected for combination drug therapy and its effects on cell viability,hepatotoxicity,and cell cycle arrest under drug-conditioned culture were *** results suggested that the combined concentration below-100μg/m L had no significant inhibitory effect on Hep G2 cell viability,and therefore Hep G2 cells maintained their drug metabolism *** the drug concentration was increased above 250μg/m L,Hep G2 cells underwent *** of metabolites by mass spectrometry proved the effective metabolism in the microchip *** dynamic,co-culture microchip successfully provided a podium for long-term observation of absorption,transport,and metabolism of combination drugs,and could be an effective in vitro simulation model for further clinical research.

关键词： intestine-liver model combination drug microchip metabolite behavior

Effect of Dai-Bai-Jie on the proliferation and migration of the A549 cells

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Chinese Chemical Letters 2020年第2期31卷 476-478页

作者： Binxin lin Xinling Lu Nan Li Ning Xu jin-ming lin Beijing Key Laboratory of Microanalytical Methods and Instrumentation Department of ChemistryTsinghua UniversityBeijing 100084China Department of Chemistry School of ScienceTianjin UniversityTianjin 300000China

Lung cancer is the most malignant tumor disease with the highest diagnosis and mortality rate in *** development of therapeutic drugs is the current research ***-Bai-Jie is a traditional medicine of the Dai nationality,which is commonly used in the treatment of decreasing swelling,alleviating pain and *** of the current researches focused on the component analysis of Dai-Bai-Jie,but few researches studied on its antitumor and pharmacological *** this study,we incubated A549 cells with different concentrations of *** cell proliferation experiment showed that the DaiBai-Jie solution inhibited the proliferation of A549 cells and caused cell *** this work,we confirmed that Dai-Bai-Jie had an inhibitory effect on the proliferation and migration of non-small cell lung cancer A549,which may be used as a novel candidate of anti-tumor therapy for lung cancer patients.

关键词： "Dai-Bai-Jie" Apoptosis Proliferation Migration Antineoplastic agents

Development of cell metabolite analysis on microfluidic platform

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Journal of Pharmaceutical Analysis 2015年第6期5卷 337-347页

作者： Luyao lin jin-ming lin Department of Chemistry Beijing Key Laboratory of Microanalytical Methods and Instrumentation Tsinghua University

Cell metabolite analysis is of great interest to analytical chemists and physiologists, with some metabolites having been identified as important indicators of major diseases such as cancer. A highthroughput and sensitive method for drug metabolite analysis will largely promote the drug discovery industry. The basic barrier of metabolite analysis comes from the interference of complex components in cell biological system and low abundance of target substances. As a powerful tool in biosample analysis, microfluidic chip enhances the sensitivity and throughput by integrating multiple functional units into one chip. In this review, we discussed three critical steps of establishing functional microfluidic platform for cellular metabolism study. Cell in vitro culture model, on chip sample pretreatment, and microchip combined detectors were described in details and demonstrated by works in five years. And a brief summary was given to discuss the advantages as well as challenges of applying microchip method in cell metabolite and biosample analysis.

关键词： Microfluidic Cell analysis Cellular metabolism Chip-mass spectrometry