RESUMO
Fluorescent rotors with aggregation-induced emission (AIE) and organelle-targeting properties have attracted great attention for sensing subcellular viscosity changes, which could help understand the relationships of abnormal fluctuations with many associated diseases. Despite the numerous efforts spent, it remains rare and urgent to explore the dual-organelle targeting probes and their structural relationships with viscosity-responsive and AIE properties. Therefore, in this work, we reported four meso-five-membered heterocycle-substituted BODIPY-based fluorescent probes, explored their viscosity-responsive and AIE properties, and further investigated their subcellular localization and viscosity-sensing applications in living cells. Interestingly, the meso-thiazole probe 1 showed both good viscosity-responsive and AIE (in pure water) properties and could successfully target both mitochondria and lysosomes, further imaging cellular viscosity changes by treating lipopolysaccharide and nystatin, attributing to the free rotation and potential dual-organelle targeting ability of the meso-thiazole group. The meso-benzothiophene probe 3 with a saturated sulfur only showed good viscosity-responsive properties in living cells with the aggregation-caused quenching effect and no subcellular localization. The meso-imidazole probe 2 showed the AIE phenomenon without an obvious viscosity-responsive property with a CâN bond, while the meso-benzopyrrole probe 4 displayed fluorescence quenching in polar solvents. Therefore, for the first time, we investigated the structure-property relationships of four meso-five-membered heterocycle-substituted BODIPY-based fluorescent rotors with viscosity-responsive and AIE properties, and among these, 1 with a CâN bond and a saturated sulfur on the meso-thiazole, potentially contributing to their corresponding AIE and viscosity-responsive properties, served as a sensitive AIE fluorescent rotor for imaging dual-organelle viscosity in both mitochondria and lysosomes.
Assuntos
Corantes Fluorescentes , Organelas , Corantes Fluorescentes/química , Viscosidade , Diagnóstico por ImagemRESUMO
Meso-substituted boron dipyrromethenes (BODIPYs) provide a potential and innovative strategy for the synergistic construction of aggregation-induced emission (AIE) probes and fluorescent rotors for monitoring cellular viscosity changes, which play critical roles in understanding the function of viscosity in its closely associated diseases. Therefore, for the first time, a BODIPY-based fluorescent probe (1) with a rotatable meso-benzothiazole group was rationally designed and synthesized, showing both good viscosity-responsive and AIE properties. Probe 1 through direct linkage with the thiazole group, showed nearly no emission in low viscous solvents; however, a strong emission at 534 nm appeared and increased gradually with the increase in viscosity, attributing to the efficient restriction of the rotatable meso-benzothiazole group. The intensity (log I534) displayed a good linear relationship with viscosity (log η) in the viscous range of 0.59-945 cP in methanol/glycerol mixtures. Interestingly, 1 showed enhanced emission at 534 nm in 70% water compared to pure acetonitrile due to the aggregation-induced inhibited rotations. Cellular imaging suggested that 1 could successfully sense lysosomal viscosity changes induced by lipopolysaccharide, nystatin, low temperature, and dexamethasone in living cells, which could be further applied in autophagy monitoring by tracing viscosity changes. As a comparison, its analogue 2 directly linking with the phenyl group showed no viscosity-responsive or AIE properties. Therefore, for the first time, we reported a meso-benzothiazole-BODIPY-based fluorescent rotor with AIE and lysosomal viscosity-responsive properties in nervous cells, which was further applied in monitoring autophagy, and this work thus could provide an innovative strategy for the design of potential AIE and viscosity-responsive probes.
Assuntos
Boro , Corantes Fluorescentes , Metanol , Glicerol , Lipopolissacarídeos , Nistatina , Lisossomos , Benzotiazóis , Acetonitrilas , Solventes , Autofagia , Água , DexametasonaRESUMO
Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) has become a very efficient and sensitive trace, ultratrace, and surface analytical technique for the in situ study of the concentration and distribution of the elements in life sciences with high spatial resolution. It is being used more and more frequently in biological, medical materials and protein research, which will lead to a better understanding of physiology and pathology process in cells and tissues. The present review mainly introduces the strategies of combination of gel electrophoresis (GE) with LA-ICP-MS for the quantification of trace elements in proteins, including the proteins separation, elements detection and calibration methods. The paper emphasizes the basic conditions of the proteins separation, focusing on the stability of proteins during GE and the treatment methods of staining and drying of the gel to enable successful detection of the elements by LA-ICP-MS. In addition, the application of GE-LA-ICP-MS in phosphoproteins, selenoproteins and metal-binding proteins is introduced in detail. The prospects and challenge for this technique are discussed as well for further study.
Assuntos
Eletroforese , Espectrometria de Massas , Proteínas/química , Oligoelementos/química , CalibragemRESUMO
Two novel meso-CF3 BODIPY-based fluorescent rotors have been rationally prepared and found to sensitively respond to viscosity in living cells with a fluorescence "turn-on" effect, attributed to the special restricted rotation of meso-CF3 group in viscous environments. Interestingly, a monostyryl probe with one cationic group exhibits good mitochondrial localization and AIE property.
Assuntos
Compostos de Boro/química , Corantes Fluorescentes/química , Mitocôndrias/metabolismo , Viscosidade , Cátions , Células HeLa , Humanos , Estrutura Molecular , Espectrometria de FluorescênciaRESUMO
Hypochlorous acid/hypochlorite (HOCl/ClO-) plays important roles in killing bacterial and causing damage to living tissues, and its abnormal levels could lead to many diseases. Although great efforts have been devoted, fluorescent probes for HOCl/ClO- with near-infrared fluorescence, good selectivity/sensitivity, and low background are still important and urgent. In this work, a novel double-bond-linked TCF-aza-BODIPY-based near-infrared fluorescent probe (3) was rationally designed, successfully prepared, and applied for sensing HOCl/ClO- in both solutions and living RAW264.7 cells, showing good selectivity and fluorescence "turn-on" phenomenon at 670 nm with low background. The limit of detection towards ClO- was determined to be 0.36 µM through the linear fluorescence changes at 670 nm in a broad ClO--concentration range of 0-150 µM. Furthermore, the sensing mechanism was investigated by mass spectrometry and compared with 1, suggesting that the remarkable spectroscopic changes could be ascribed to the oxidization of the double bond to the aldehyde group, accompanied with the leaving of the TCF group. Confocal imaging experiments also confirmed the remarkable intracellular fluorescence enhancements through incubation of ClO- and phorbol ester 12-myristate 13-acetate (PMA) in RAW264.7 cells. Therefore, for the first time, we reported a near-infrared TCF-aza-BODIPY-based fluorescent probe for highly sensitive and fluorescence "turn-on" detection of both exogenous and endogenous HOCl in living RAW264.7 cells through the quick oxidation of a conjugated double bond.
Assuntos
Corantes Fluorescentes , Ácido Hipocloroso , Compostos de Boro/química , Corantes Fluorescentes/química , Ácido Hipocloroso/análise , Microscopia de Fluorescência/métodosRESUMO
Hydrogen peroxide (H2O2) plays vital roles in oxidative stress and signal transduction in living organisms, and its abnormal levels could be linked to many diseases. Despite numerous efforts spent, it is still urgent and of high importance to develop better H2O2 probes with good selectivity, high sensitivity and low backgrounds. To this end, a novel boron dipyrromethene (BODIPY)-based fluorescent probe with an electron-withdrawing methylenemalononitrile at the meso position has been rationally designed, successfully synthesized and investigated for detection of H2O2 in aqueous solutions and living cells, which exhibited high selectivity and sensitivity, fluorescent "turn-on" phenomenon at 540 nm, and ratiometric changes from 506 to 540 nm. Upon exposure to H2O2, a strong fluorescent emission at 540 nm appeared and the corresponding quantum yields changed from 0.009 to 0.13. The detection limit towards H2O2 was calculated to be 31 nM by the linear fluorescence change at 540 nm in the H2O2-concentration ranging from 2 to 10 µM. This probe was applicable in a pH range from 6 to 10. Meanwhile, the sensing mechanism was also confirmed by the 1H NMR and mass spectrometry, suggesting that the above changes might be ascribed to the quick addition and oxidization of the double bond. Furthermore, confocal imaging results also showed great enhancement of intracellular fluorescence upon exposure to H2O2 and PMA in RAW264.7 cells, unambiguously confirming its great potentials as a fluorescent probe for highly sensitive detection of both exogenous and endogenous H2O2 in living cells.
Assuntos
Compostos de Boro/química , Corantes Fluorescentes/química , Peróxido de Hidrogênio/análise , Nitrilas/química , Imagem Óptica , Animais , Relação Dose-Resposta a Droga , Corantes Fluorescentes/síntese química , Células HEK293 , Humanos , Camundongos , Estrutura Molecular , Células RAW 264.7 , Relação Estrutura-AtividadeRESUMO
A novel near-infrared-emitting aza-BODIPY-based fluorescent probe with two tellurium atoms at two upper benzyl rings has been prepared and explored for its fluorescent sensing properties towards hypochlorous acid/hypochorite (HClO/ClO-), which showed high selectivity and absolutely fluorescent "turn-on" phenomenon at 738 nm. The fluorescence of this probe was sufficiently quenched due to photoindued electron transfer by two tellurium atoms. Upon exposure to HClO/ClO-, a strong near-infrared emission at 738 nm appeared with fluorescence quantum yields changing from 0 to 0.11. This remarkable fluorescence change was ascribed to the oxidation of both electron-rich tellurium atoms. The detection limit of this probe towards HClO/ClO- was calculated to 0.09 µM in acetonitrile aqueous solution by the linear fluorescence change at 738 nm in the HClO/ClO--concentration range of 0-30 µM. Interestingly, this probe was found to be applicable in a broad pH range (2-10). Meanwhile, the oxidized probe could be further responsive to biothiols with substantial fluorescence disappearance. The bioimaging experiments in RAW264.7 cells showed the appearance of intracellular near-infrared fluorescence after addition of HClO/ClO- and PMA, and the fluorescence could also be reversed to be silenced by further introduction of GSH, confirming its potential application for exogenous and endogenous detection of HClO/ClO- in living cells.
Assuntos
Corantes Fluorescentes , Ácido Hipocloroso , Compostos de Boro , Microscopia de FluorescênciaRESUMO
A new near-infrared (NIR)-emitting aza-boron-dipyrromethene dye with two electron-donating amino groups at 1- and 7-positions has been prepared via several steps of reactions. This probe showed a NIR absorption at 748 nm with an obvious shoulder peak at 634 nm in CH3CN/H2O. Interestingly, a NIR fluorescence emission at 843 nm was observed with a large Stokes shift of 95 nm. This novel NIR-emitting aza-boron-dipyrromethene dye was further investigated as a Hg2+-sensing fluorescent probe, which selectively bound to Hg2+, showing a blue-shifted and sharp absorption band at 695 nm with the disappearance of the shoulder peak at 634 nm. Correspondingly, the color change could be easily seen from blue to green. Interestingly, the emission exhibited an absolutely "turn-on" peak at 725 nm with a significant blue shift by 118 nm (from 843 to 725 nm), due to the efficient inhibition of the intramolecular-charge-transfer process arising from two amino groups. This probe was finally introduced to Hela cells, showing a "OFF-ON" NIR emission upon exposure to Hg2+. The overall results confirmed that this novel NIR-emitting aza-boron-dipyrromethene fluorescent probe with a large Stokes shift could serve as a colorimetric and fluorescent "turn-on" sensor for Hg2+ in both solutions and living cells.
Assuntos
Corantes Fluorescentes , Mercúrio , Boro , Compostos de Boro , Células HeLa , Humanos , Porfobilinogênio/análogos & derivadosRESUMO
A simple, rapid and inexpensive fluorimetric method for simultaneously determining ASA and SA in a single scan of the co-existing specimen had been established. When the wavelength interval (delta lambda) is 80 nm for scaning, the amounts of ASA and AS could be quantitatively determined respectively. The fluorescence intensity was linearly related to the aspirin concentration in the range of 4.0 x 10(-6)-1.0 x 10(-4) mol x L(-1) with correlation coefficient 0.9949; the fluorescence intensity was linearly related to the salicyclic acid concentration in the range of 8.0 x 10(-7)-1.0 x 10(-4) mol x L(-1) with correlation coefficient 0.9975, the determination limit was 4.0 x 10(-7) mol x L(-1). The method is simple, rapid and inexpensive, it can be used for medicament analysis.
Assuntos
Aspirina/análise , Salicilatos/análise , Espectrometria de Fluorescência/métodos , Reprodutibilidade dos TestesRESUMO
The soyabean isoflavones genistein (GEN) and daidzein (DA) are popular presented in diet. Isoflavones have a variety of biological activities including antioxidant and anticancer properties. On account of its antioxidant activity, isoflavones might protect cancer cells from free radical damage in photodynamic (PDT) during which reactive oxygen species (ROS) production was stimulated leading to irreversible tumor cell injury. In this study, the influence of GEN and DA on K562 cells in 5-aminolevulinic acid (ALA)-based PDT was demonstrated. The results showed that GEN inhibited cell proliferation and enhance cell apoptosis, lipid peroxidation, and DNA damage in ALA-PDT on K562 cells. However, DA did not enhance cell apoptosis, lipid peroxidation, and DNA damage in ALA-PDT. In conclusion, the results suggested that soy consumption during PDT did not decrease the effectiveness of cancer therapy on malignant cells.
Assuntos
Ácido Aminolevulínico/uso terapêutico , Genisteína/farmacologia , Isoflavonas/farmacologia , Células K562/efeitos dos fármacos , Fotoquimioterapia , Antioxidantes/farmacologia , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Dano ao DNA/efeitos dos fármacos , Dieta , Humanos , Peroxidação de Lipídeos/efeitos dos fármacos , Fármacos Fotossensibilizantes/farmacologiaRESUMO
BACKGROUND: Histone deacetylase (HDAC) inhibitors represent a novel class of therapeutic agents with antitumor activity currently in clinical development. In this study, we tested the biological effects of the HDAC inhibitor MS-275 in various pre-clinical prostate cancer models both in'vitro and in vivo. METHODS: In vitro cell proliferation XTT assay and protein expression analysis by Western blot were performed. In vivo tumor growth assessment in subcutaneous, orthotopic, and transgenic mouse models were conducted. RESULTS: MS-275 significantly upregulated histone H3 acetylation and p21 gene expression in human prostate cancer cell lines. MS-275 exerted growth arrest in PC-3 and LNCaP cells, and induced cell death in DU-145 cells. Prostate specific antigen protein levels were increased by MS-275 in LAPC4 cell line. In vivo, MS-275 inhibited the growth of DU-145, LNCaP, and PC-3 in subcutaneous xenografts. MS-275 had also a significant inhibition of PC-3 cells growth in a mouse intratibial model. Molecular analysis showed increased histone acetylation and p21 expression in tumor samples from MS-275-treated mice. In transgenic adenocarcinoma of mouse prostate (TRAMP) mice, long-term treatment of MS-275 slowed the progression of prostate carcinomas with significant reduction in cell proliferation. CONCLUSIONS: Taken together, these data support the clinical testing of MS-275 for the treatment of prostate cancer.