研究目的
To study the effect of stilbene derivative c-TAB on cardiomyocyte layers and voltage-gated ion channels in cardiac cells, focusing on its ability to reversibly and irreversibly inhibit excitation, and its lower toxicity compared to AzoTAB.
研究成果
c-TAB can reversibly inhibit excitability in cardiac cells in both trans- and cis- forms, with inhibition reversible by washout. UV irradiation of trans-c-TAB leads to permanent inhibition. The effect is due to modulation of voltage-gated ion channels, with reduced toxicity compared to AzoTAB, suggesting potential applications in cardiac ablation and anesthesia.
研究不足
The study is limited to in vitro experiments on neonatal rat cardiomyocytes; in vivo applications and delivery methods for c-TAB are not addressed. The irreversible inhibition requires UV light, which may have practical constraints in clinical settings. Toxicity, while lower than AzoTAB, is still present and needs further optimization.
1:Experimental Design and Method Selection:
The study involved optical mapping and patch-clamp techniques to monitor excitation waves and record ion currents in neonatal rat ventricular cardiomyocytes (NRVMs). Photosomerization was induced using UV and blue light.
2:Sample Selection and Data Sources:
NRVMs were isolated from 1-2-day-old Wistar rats using enzymatic dissociation. Cells were cultured and used for experiments.
3:List of Experimental Equipment and Materials:
Included CMOS camera, patch-clamp setup, spectrophotometer, NMR spectrometer, UV-LED modules, and various reagents such as c-TAB, Fluo-4 dye, and culture media.
4:Experimental Procedures and Operational Workflow:
Cells were treated with c-TAB, exposed to light for isomerization, and subjected to electrophysiological recordings and optical imaging. Data were collected on ion currents, action potentials, and excitation propagation.
5:Data Analysis Methods:
Data were analyzed using Clampfit, SigmaPlot, and ImageJ software, with statistical significance assessed using Student's t-test.
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CMOS camera
pco.1200hs
PCO AG
Acquiring images for optical mapping of excitation waves
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Image intensifier unit
C8600
Hamamatsu
Enhancing fluorescence signals for imaging
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Microscope
MVX10
Olympus
Viewing and imaging cell cultures
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UV-LED module
LC-L2
Hamamatsu
Inducing trans-to-cis isomerization with UV light
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UV-LED
LC-L1V3
Hamamatsu
Light source for photoisomerization
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LED head unit
L11921
Hamamatsu
Applying UV light to experimental chamber
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Spectrophotometer
U0080D
Hitachi
Recording absorption spectra of c-TAB isomers
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Patch pipettes
BF150-86-10
Sutter Instrument
Used in patch-clamp experiments for electrophysiological recordings
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Laser power meter
Nova II P/N 7Z01550
Ophir Spiricon Europe GmgH
Measuring power density of UV light
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NMR spectrometer
Varian Inova 500 WB
Varian
Recording NMR spectra for molecular analysis
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