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Visualizing Nitric Oxide in Mitochondria and Lysosomes of Living Cells with N-Nitrosation of BODIPY-based Fluorescent Probes
摘要： Nitric oxide (NO), a ubiquitous gasotransmitter which plays critical roles in cardiovascular, nervous, and immune systems related diseases, is closely related in the physiological and pathological processes of mitochondria and lysosomes. Thus, monitoring NO in mitochondria or lysosomes is very meaningful for NO related chemical biology. Herein, we rationally designed four NO probes, BDP-NO, Mito-NO-T, Mito-NO and Lyso-NO, based on BODIPY dye substituted at meso position with 5-amino-2-methoxy-phenyl scaffold. These four probes all showed fast fluorescence off-on response toward NO with excellent selectivity and high sensitivity with the detection limit of BDP-NO to reach 5.7 nM. We introduced triphenylphosphonium and morpholine moieties onto BODIPY scaffold respectively to enable organelle-targetability. MTT and flow cytometry assay demonstrated that the probes exhibited low cytotoxicity, which was beneficial to the biological application in living cells. Confocal fluorescence microscopy experiments confirmed excellent mitochondria targeting for Mito-NO and lysosome-targeting with Lyso-NO for the detection of NO in living cells.
关键词： Fluorescent probes,Mitochondria-targeted,Lysosomes-targeted,BODIPY,Nitric oxide
更新于2025-11-21 11:08:12
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Primary and secondary photochemical transformations of biologically active precursor - Nitro-Nitrosyl ruthenium complex
摘要： Photolysis of [Ru(II)NOPy2(NO2)2OH] (A) complex was studied under blue light (445 nm) irradiation. The primary photo-process results in the formation of nitric oxide (NO) and a paramagnetic RuIII compound, the latter was detected by EPR technique. The quantum yield (6–11%) of primary photolytic process was determined from the evolution of UV–vis spectra in different solvents (water, ethanol, dimethyl sulfoxide and acetonitrile). The secondary processes compete with the NO-release and lead to a variety of ruthenium forms including new nitrosyl forms. Namely, processes of hydroxyl protonation, pyridine photocleavage and nitro – nitrito bond isomerization take a place. HPLC combined with IR and 15N – NMR spectroscopies clarified the composition of the new forms and their transformations during the photolysis. Supporting EPR and IR DFT calculations confirmed the structure of photoproducts. The oxidation of NO in the secondary processes significantly influences on the total amount of nitric oxide release and reaction routes.
关键词： Nitrosyl ruthenium,Photocleavage,Photochemistry,Nitric oxide release
更新于2025-09-23 15:23:52
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A molecular hybrid producing simultaneously singlet oxygen and nitric oxide by single photon excitation with green light
摘要： Combination of photosensitizers (PS) for photodynamic therapy with NO photodonors (NOPD) is opening intriguing horizons towards new and still underexplored multimodal anticancer and antibacterial treatments not based on “conventional” drugs and entirely controlled by light stimuli. In this contribution, we report an intriguing molecular hybrid based on a BODIPY light-harvesting antenna that acts simultaneously as PS and NOPD upon single photon excitation with the highly biocompatible green light. The presented hybrid offers a combination of superior advantages with respect to the other rare cases reported to date, meeting most of the key criteria for both PSs and NOPDs in the same molecular entity such as: (i) capability to generate 1O2 and NO with single photon excitation of biocompatible visible light, (ii) excellent 1O2 quantum yield and NO quantum efficiency, (iii) photogeneration of NO independent from the presence of oxygen, (iv) large light harvesting properties in the green region. Furthermore, this compound together with its stable photoproduct, is well tolerated by both normal and cancer cells in the dark and exhibits bimodal photomortality of cancer cells under green light excitation due to the combined action of the cytotoxic 1O2 and NO.
关键词： Green light,Nitric oxide,Multimodal therapy,Singlet oxygen,BODIPY,Photodynamic therapy
更新于2025-09-23 15:22:29
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Fluorescent Calixarene Scaffolds for NO Detection in Protic Media
摘要： Here, we present the first fluorescent water-soluble conjugated calixarene scaffolds that are capable of detection of NO gas. Two different scaffolds, one based on a 5,5’-Bicalixarene structure and its isomer bearing two distantly conjugated calixarene moieties, have been synthesized. While the fluorescence of both isomers is quenched upon either passing of NO gas or generating it in-situ from diethylamine NONOate, the bicalixarene-based scaffold showed a significantly stronger response. We also present an example of a dye encapsulation strategy to achieve the detection of NO at longer wavelengths than in the parent calixarene host. Finally, a conjugated polymer bearing 5,5’-Bicalixarene scaffold has also been prepared and demonstrated enhanced sensitivity compared with the monomer due to the molecular wire effect.
关键词： conjugated systems,nitric oxide,host-guest chemistry,fluorescent probes,Calixarene
更新于2025-09-23 15:22:29
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A rhodamine-deoxylactam based fluorescent probe for fast and selective detection of nitric oxide in living cells
摘要： Nitric oxide (NO) plays vital roles in many physiological process and is closely related to many diseases. So far, a number of fluorescent probes have been constructed for the detection of NO successfully. However, the probes still suffer from long-time reaction and limited selectivity. Herein, a fluorescent probe named dRB-OPD is synthesized and used to recognize NO. The probe contains a deoxy-rhodamine B as fluorophore and o-phenylenediamino as reaction site. dRB-OPD shows fast response to NO within 40 seconds with 170-fold fluorescence enhancement. Moreover, the probe shows high selectivity towards NO over dehydroascorbic acid (DHA), ascorbic acid (AA), and methylglyoxal (MGO). Particularly, the probe can avoid the serious interference from cysteine (Cys) found in the rhodamine lactam-based fluorescent NO probes (RB-OPD). In addition, the probe is applied for the detection of exogenous and endogenous NO in the HepG2 and RAW 264.7 cells with satisfactory results.
关键词： Cell imaging,Fluorescent probe,Nitric oxide,High selectivity,Fast response
更新于2025-09-23 15:22:29
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Evaluation of nitric oxide laser-induced fluorescence thermometry techniques in a hypersonic boundary layer
摘要： Nitric oxide planar laser-induced fluorescence was performed to measure the wall-normal distribution of static temperature through a hypersonic boundary layer. A 10-degree half-angle wedge model was oriented at a 5-degree angle of attack in the NASA Langley 31-in Mach 10 facility, resulting in a 5-degree flow turning angle and an edge Mach number of 7.6. Nitric oxide was seeded through a spanwise slot into the boundary layer upstream of the imaging region and was excited with a pulsed ultraviolet planar laser sheet. The laser was spectrally scanned across six fluorescence transitions in the (0, 0) band of the A2Σ+–X2Π system. Eighteen thermometry methods were assessed through comparison to predictions of the temperature field from computational fluid dynamics simulations. The effect of spectral resolution and laser linewidth on measurement uncertainty was also investigated. The most accurate technique was spectral peak thermometry, which achieved an accuracy of ± 31.6 K (12.6% error relative to CFD temperature). The spectral peak thermometry technique required a minimum spectral resolution between 0.074 and 0.102 cm?1 to extract meaningful temperature information from the spectra and a maximum laser linewidth of 0.49 cm?1.
关键词： Laser linewidth,Thermometry,Spectral resolution,Planar laser-induced fluorescence,Hypersonic boundary layer,Nitric oxide
更新于2025-09-23 15:21:01
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Nitric oxide sensors using nanospiral ZnO thin film deposited by GLAD for application to exhaled human breath
摘要： ZnO is a promising gas sensing material for its excellent gas sensing response characteristics and long-term stability. Moreover, the improvement in the sensitivity and response speed of ZnO gas sensors can be achieved by the nanostructure fabrication. This paper proposes a facile method to deposit ZnO nanospirals using glancing angle deposition (GLAD) for application in nitric oxide (NO) sensors. ZnO nanospirals with porous characteristics have larger relative surface area and more active surfaces, compared with dense ZnO thin film. A sensor using nanospiral ZnO film shows a response factor of 16.9 to 100 ppb NO at 150 °C in 40% RH, which is 3 times larger than that of the sensor using dense ZnO film. Such a ZnO nanospiral sensor system can detect NO as low as 10 ppb which is below the NO concentration (>30 ppb) in exhaled breath of patients with asthma. The effects of working temperature and humidity on the sensor performance were investigated systematically in this work. Moreover, the sensor response showed a good selectivity to NO and high stability as the time increased up to 24 days. NO gas sensing mechanism was discussed in detail and nanospiral ZnO film sensors are promisingly applicable for exhaled human breath application compared with some other NO sensors.
关键词： exhaled breath,nitric oxide,GLAD,nanospiral,gas sensor,ZnO
更新于2025-09-23 15:19:57
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Organic semiconducting nanoprobe with redox-activatable NIR-II fluorescence for <i>in vivo</i> real-time monitoring of drug toxicity
摘要： An activatable organic semiconducting nanoprobe that specifically turns on its second near-infrared window fluorescence upon being exposed to nitric oxide stimuli was developed for in vivo, in situ, real-time and high-spatial-resolution mapping of drug-induced hepatotoxicity.
关键词： drug toxicity,nitric oxide,NIR-II fluorescence,hepatotoxicity,redox-activatable,in vivo monitoring,Organic semiconducting nanoprobe
更新于2025-09-19 17:15:36
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Assessment of tissue perfusion and vascular function in mice by scanning laser Doppler perfusion imaging
摘要： Post-occlusive reactive hyperemia (PORH) is a key feature of physiological vasomotion to appropriately match the supply/demand ratio of tissues. This adaptive mechanism is severely disturbed in endothelial dysfunction with a reduced flow-mediated dilation (FMD). Reduced PORH and FMD are powerful prognostic risk factors in cardiovascular disease. While these parameters are frequently determined in human beings, comparable methods applicable to mouse models are sparse. We aimed to evaluate the applicability and accuracy of scanning laser Doppler perfusion imaging (LDPI) to measure PORH in the mouse hindlimb. Changes in mean perfusion in response to vasoactive drugs and PORH (assessed by scanning LDPI) were compared with changes in diameter and blood flow in the femoral artery, as assessed by high-resolution ultrasound. We found that the measured LDPI signal significantly correlated with changes of inflow into the femoral artery. Vasodilation induced by administration of nitroglycerine and acetylcholine increased vessel diameter, blood flow and mean perfusion, while vasoconstriction following administration of epinephrine decreased all three parameters. PORH was induced by temporal occlusion of the femoral artery with an external cuff. During occlusion, mean perfusion decreased to a condition of zero-perfusion and release of the cuff induced an immediate increase in blood flow that was followed by femoral artery dilation driving PORH/ perfusion. Surgical removal of the femoral artery decreased mean perfusion to a zero-perfusion level and fully abolished PORH. Importantly, the measurement of the PORH response by scanning LDPI is highly reproducible as determined by repeated measurements and intra/interobserver variation analysis. Last, we found that the PORH response was dependent on nitric oxide synthase and cyclooxygenase and declined with age. Thus, we here provide novel and robust non-invasive methods to serially measure tissue perfusion at baseline and during physiological and pharmacological modulation of vasomotor tone in the hindlimb of mice. The application of these LDPI scanning and ultrasound-based methods may be useful for testing the effects of drugs affecting vasomotor function or future elucidation of mechanisms leading to vasomotor dysfunction in mice in vivo.
关键词： age-dependent endothelial dysfunction,Laser Doppler perfusion imaging,nitric oxide synthase,hindlimb,reactive hyperemia
更新于2025-09-19 17:13:59
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A ruthenium nitrosyl-functionalized magnetic nanoplatform with near-infrared light-controlled nitric oxide delivery and photothermal effect for enhanced antitumor and antibacterial therapy
摘要： Developing spatiotemporal-controlled nitric oxide (NO) delivery nanoplatform is highly desirable for its biological applications as tumor inhibitor and antibacterial agent. In this study, a novel multifunctional magnetic nanoplatform {Fe3O4@PDA@Ru-NO@FA} (1) was developed for the near-infrared (NIR) light-controlled release of NO, in which a ruthenium nitrosyl (Ru-NO) donor and folic acid (FA)-directing group were covalently functionalized onto Fe3O4@PDA. Nanoplatform 1 preferentially accumulated in folate receptor-overexpressing cancer cell lines and magnetic field-guided tumor tissue, instantly released NO, and simultaneously produced a prominent photothermal effect upon 808 nm NIR light irradiation, leading to remarkable in vitro and in vivo antitumor efficacy. When nanoplatform 1 was treated only once, the potential MRI contrast agent was sufficient to significantly inhibit or eliminate the tumor tissues in living mice, thus offering opportunities for future NO-involved multimodal cancer therapy. In addition, a NO delivery nanoplatform {Fe3O4@PDA@Ru-NO} was imbedded in the matrix of chitosan (CS)-poly(vinyl alcohol) (PVA) material to develop a hybrid thermosensitive CS-PVA/NO hydrogels. The CS-PVA/NO hydrogels demonstrated mild (<150 mW cm-2) NIR light-controlled NO delivery and thus produced efficient antibacterial effect for both Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus. Therefore, these hydrogels feature potential as antibacterial dressings for treating wound bacterial infection.
关键词： nitric oxide delivery,magnetic nanoplatform,ruthenium nitrosyl donor,antitumor and antibacterial therapy,thermosensitive hydrogel
更新于2025-09-12 10:27:22