CHEMOSPHERE
·
2022
·
IF 8.943
Xupeng Wang,Bing Gong,Erkai He,Willie J.G.M. Peijnenburg,Hao Qiu
Awareness of toxicokinetics at the subcellular level is crucial to deciphering the underlying intoxication processes of metal(loid)s, although this information is often lacking. Here, the toxicokinetics of two non-essential metal(loid)s (Cd and As) and one essential metal (Zn) in both the whole body and subcellular fractions of earthworm ( Eisenia fetida ) were assessed. Earthworms were exposed to natural soils originating from a gradient of metal(loid) pollution for 14 days followed by a 14-day elimination phase in clean soil. Clearly distinct toxicokinetic patterns were found in the earthworms according to the metal(loid) considered. An obvious concentration-dependent increase was observed in earthworms or subcellular compartments where no equilibrium was reached (with slow or no elimination) for Cd and As throughout the experiment. As for Zn, the earthworms were able to retain a steady-state concentration of Zn in its body or each fraction without a clear intake behavior via the dynamic trade-off between uptake and elimination at different pollution levels. These differences in toxicokinetics at the subcellular level supported the observed differences in bioaccumulation patterns and were indicative of the strategy by which non-essential and essential elements are handled by earthworms. Notably, the concentration of Cd and As in subcellular compartments showed the same pattern as for Zn in the order of cellular cytosol > cellular debris > metal-rich granules, which might be associated with the binding of non-essential/essential elements with metallothionein enriched in the cytosol. Our findings enhance the understanding of the underlying mechanisms for metal(loid) accumulation kinetics in earthworms from the perspective of subcellular partitioning, and will be beneficial for accurate risk assessment of Cd, As, and Zn.
研究领域:
环境科学
·
毒理学
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DOI
PubMed
高影响因子
Acupuncture and Herbal Medicine
·
2024
·Vol. 4
·期 2
·243
·
IF 0
Wu Jianzhi,Ma Zhi,Zhang Yinhao,Duan Shuni,Sun Rong,Liu Runping,Li Yijie,Li Xiaojiaoyang
Objective: Drug-induced liver injury (DILI) is the leading cause of acute liver failure and poses a significant challenge to human health. Rhubarb ( Rheum officinale Baill . DaHuang) has been clinically used for its heat-clearing and diuresis-promoting effects. However, its toxic effects on different organelles in the liver require further validation. Methods: We analyzed the potential targets affecting hepatotoxicity in rhubarb and the potential damage relationship with five major organelles, including microsomes, mitochondria, endoplasmic reticulum (ER), Golgi apparatus (GA), and lysosomes through Integrated Traditional Chinese Medicine (ITCM)/HERB databases and network pharmacology. We isolated and purified different organelles, incubated them with different fractions and monomers of rhubarb in an adenosine triphosphate (ATP) culture system and examined the structural and functional changes in the organelles using particle size analysis and molecular biological experiments to investigate whether rhubarb affects the damage and rupture of major organelles in the liver. Results: By combining virtual predictions and experimental verification, our research confirmed that emodin isolated from the anthraquinone of rhubarb, catechin in the tannins of rhubarb, and palmitic acid in the organic acids of rhubarb caused the most significant functional and structural damage to the representative organelles. Among all the monomeric compounds, emodin caused the most damage to the microsome, mitochondria, ER, and lysosome; catechin induced microsome and GA damage; and palmitic acid caused the most damage to microsomes and GA in the liver, suggesting that rhubarb components may exert hepatotoxicity through multi-organelle injury. Conclusions: Our findings revealed that rhubarb has varying degrees of damaging effects on different organelles, which in turn affects cellular life activities by impairing organelle morphology and function. This study provides a theoretical basis and technical support for a refined analysis of the toxic components and targets of rhubarb. Export
研究领域:
中药学
·
药理学
·
细胞生物学
·
生物化学
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DOI
JOURNAL OF FLUORESCENCE
·
2022
·Vol. 33
·期 3
·923-932
·
IF 2.525
Zhang Zhi-Hao,Yu Qing,Zhang Gang,Peng Hai-Yan,Sun Ru,Xu Yu-Jie,Ge Jian-Feng
Two chromenoquinoline-based fluorescent probes 1a-b have been synthesized and investigated. Photofading behaviors of compounds 1a-b showed that at least 89% absorption remained after 6 h irradiating, meanwhile, many of ions and amino acids had negligible impacts on their fluorescence intensity, which meant they had excellent photostability and selectivity. Probes 1a-b exhibited strong absorption and emission in organic solvents with large fluorescence quantum yields, even in water probe 1a still had a relatively large fluorescence quantum yield (20%). Combined with DFT calculation, the influence of alkylation on optical properties of 1b was elucidated. In addition, the fluorescence intensity of probe 1b with red emission enhanced by 5.4-fold and 5.3-fold after DNA and RNA added, and the fluorescence quantum yield increased from 3% to 17% and 14%, respectively, but the neutral molecule 1a had no response to nucleic acid. Furthermore, confocal microscopy imaging of probes 1a-b showed that 1a targeted lipid droplets while the methylated probe 1b to nucleus in living HeLa cells. The results indicated that the subcellular targeting zone could be changed by alkylation of nitrogen atom on chromenoquinoline-based conveniently, which provided a new idea for designing and synthesizing new subcellular labeled probes.
研究领域:
生物化学
·
分子生物学
·
药物化学
✓ 已验证
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2018
·
IF 6.042
Ruo-Can Qian,Jian Lv,Yi-Tao Long
Recently, a variety of strategies have been developed for single-cell detection. However, the precise probing of the given area at single-cell level is still a challenge. Here, we put forward a rapid and targeted imaging approach for the mapping of subcelluar domains, which realizes the precise injection of multifluorescence into a single living cell via an ultrasmall quartz capillary nanopipette (∼100 nm) and can successfully transport different fluorescent probe molecules to the pointing subcellullar area around the tip in the cytoplasm within 20 s. This method is also applied for monitoring the loss of intracellular mitochondrial membrane potential under the treatment of metformin in a single MCF-7 breast cancer cell. The major driven force in the nanopipette, electroosmotic flow, is evaluated by a theory calculation method and finite element simulations, and the solution indicates a confined solute distribution profile around the tip within the working range. Overall, the nanopipette approach realizes the precise and simultaneous delivery of multiple probe molecules into the single living cell through the electroosmotically modulated, nondestructive, and one-step injection, which is especially powerful and convenient for multichannel single-cell imaging and monitoring, indicating favorable potential for understanding, monitoring, and controlling the biological processes from the single cell to subcellular organelles.
研究领域:
细胞生物学
·
生物物理学
·
分析化学
✓ 已验证
DOI
PubMed
高影响因子
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY
·
2019
·
IF 4.527
Wenli Huang,Zhenqing Bai,Jie Jiao,Honglin Yuan,Zhian Bao,Shaoning Chen,Meihai Ding,Zongsuo Liang
Coptis chinensis Franch., is a widely used medicinal plant in China. This plant is often contaminated by cadmium (Cd) and render health risk to human consumers. Understanding distribution of Cd and its chemical forms is important to evaluate accumulation of the metal and its detoxification mechanisms in this plant. Since few studies have focused on this aspect, we used laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) to spatially locate Cd in rhizome cross-sections, and ICP-MS to analyze the Cd subcellular distribution and the chemical forms of Cd in different tissues. Rhizome bioimaging results showed that Cd was distributed predominantly within the periderm, cortex, pith, and root trace vascular bundle. The LA-ICP-MS results suggested that Ca 2+ channels might be a pathway for Cd entry into the plant. Subcellular distribution data indicated that most of Cd was associated with the cell wall (41.8–77.1%) and the soluble fraction (14.4–52.7%) in all tissues. Analysis of chemical forms revealed that majority Cd existed in less mobile and less toxic forms in all tissues, and P could convert to insoluble phosphate with Cd to moderate Cd toxicity. The new understanding of Cd accumulation and detoxification might provide novel strategies for reducing the levels of Cd in C. chinensis Franch., thereby mitigating its potential transfer to humans and providing a theoretical basis for evaluating the Cd status in other medicinal plants. Further, our findings might provide a basis for establishing a reasonable Cd limit level of traditional Chinese medicinal materials.
研究领域:
植物学
·
环境科学
·
药用植物研究
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2023
·
IF 8.4
Ning Xu,Qinglong Qiao,Yi Tao,Pengjun Bao,Yinchan Zhang,Wei Zhou,Jin Li,Zhifeng Li,Zhaochao Xu
Subcellular pH plays a key role to modulate protein structures or specific active sites to impact diverse biological processes. The heterogeneity and high dynamics of intracellular pH require probes that can detect pH in the vicinity of the protein and have a wide pH response range. Here, we reported a protein proximity ligation fluorescent probe for super-resolution imaging of subcellular pH dynamics. Relying on SNAP-tag protein labeling technology, the pH-sensitive fluorophore BG-NDM based on photo-induced electron transfer mechanism was localized to the adjacent positions of the target protein in different subcellular organelles. The equilibrium between the non-fluorescent aggregate and the fluorogenic monomer enabled BG-NDM to have the ability of wash-free super-resolution imaging in live cells. And the wide pH response ranges from pH 6 to pH 12 made the probe have the ability of recognizing pH dynamics. The pH changes around functional proteins during mitophagy, cellular alkalosis and acidosis were revealed. Through long-term super-resolution imaging, we could further record the pH dynamics around mitochondrial proteins COX8A during mitochondrial contacts, fusion and fission at the resolution of a single mitochondria.
研究领域:
生物化学
·
细胞生物学
·
分子影像学
✓ 已验证
DOI
高影响因子
ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH
·
2020
·Vol. 28
·期 11
·13955-13969
·
IF 3.056
Wang Juncai,Chen Xunfeng,Chu Shaohua,Hayat Kashif,Chi Yaowei,Zhi Yuee,Zhang Dan,Zhou Pei
Solanum nigrum is a well-documented cadmium (Cd) hyperaccumulator; however, its Cd-induced tolerance capability and detoxification mechanism remain elusive. Hence, a short-term hydroponic experiment was performed in a multiplane glasshouse to determine the influence of Cd toxicity on subcellular distribution, chemical forms, and the physiological responses of cell wall towards Cd stress in a 4-week-old plant. The experiment was conducted following completely randomized design (CRD) with five treatments ( n = 4 replicates). The results showed that Cd stress showed dose-dependent response towards growth inhibition. The subcellular distribution of Cd in S. nigrum was in the order of cell wall > soluble fractions > organelles, and Cd was predominantly extracted by 1 M NaCl (29.87~43.66%). The Cd contents in different plant tissues and cell wall components including pectin, hemicellulose 1 (HC1), hemicellulose 2 (HC2), and cellulose were increased with the increase in Cd concentrations; however, the percentage of Cd concentration decreased in pectin and cellulose. Results of the polysaccharide components such as uronic acid, total sugar contents, and pectin methylesterase (PME) activity showed Cd-induced dose-dependent increase relative to exposure Cd stress. The pectin methylesterase (PME) activity was significantly ( p < 0.05) enhanced by 125.78% at 75 μM Cd in root, 105.78% and 73.63% at 100 μM Cd in stem and leaf, respectively. In addition, the esterification, amidation, and pectinase treatment of cell wall and Fourier transform infrared spectroscopy (FTIR) assay exhibited many functional groups that were involved in cell wall retention Cd, especially on carboxyl and hydroxyl groups of cell wall components that indicated that the –OH and –COOH groups of S. nigrum cell wall play a crucial role in Cd fixation. In summary, results of the current study will add a novel insight to understand mobilization/immobilization as well as detoxification mechanism of cadmium in S. nigrum .
研究领域:
植物学
·
环境科学
·
毒理学
✓ 已验证
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2020
·
IF 6.785
Shushu Ding,Meina Li,Haoyang Gong,Qing Zhu,Guoyue Shi,Anwei Zhu
Hydroxyl radical (•OH) is an essential reactive oxygen species involved in critical cell functions. However, the mechanisms controlling its subcellular localization and intracellular level during health and disease remain poorly understood. This is due to the challenge of detecting •OH that are highly reactive and consequently short-lived (in vivo half-life of ∼10–9 s). Herein, we present tungsten nanoelectrodes functionalized with stable 1-hexanethiol (HAT) for selective and sensitive detection of •OH at the subcellular level via the destruction of the self-assembled monolayer of HAT on the nanoelectrode tip. Taking advantage of the ultrasmall nanotip and the super mechanical toughness, the tungsten nanoelectrode could easily penetrate a single living cell without inducing any observable damage. Controlled by a high precision micromanipulator, the •OH level in RAW 264.7 murine macrophages under amyloid β (Aβ) induced oxidative stress were first investigated by the nanoelectrodes at the subcellular level. Moreover, the results revealed the cordycepin-mediated cytoprotection of macrophages through modulation of PI3K/Akt pathway activity and introduction of heme oxygenase-1 (HO-1). We believe that the developed nanoelectrochemical method has shown great capacities for the study of potential drugs for therapeutic intervention of Alzheimer’s disease.
研究领域:
生物化学
·
医学
·
纳米技术
✓ 已验证
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2022
·
IF 8.008
Jian Lv,Xiao-Yuan Wang,Xin-Yue Zhou,Da-Wei Li,Ruo-Can Qian
Targeted delivery and labeling of single living cells in heterogeneous cell populations are of great importance to understand the molecular biology and physiological functions of individual cells. However, it remains challenging to perfuse fluorescence markers into single living cells with high spatial and temporal resolution without interfering neighboring cells. Here, we report a single cell perfusion and fluorescence labeling strategy based on nanoscale glass nanopipettes. With the nanoscale tip hole of 100 nm, the use of nanopipettes allows special perfusion and high-resolution fluorescence labeling of different subcellular regions in single cells of interest. The dynamic of various fluorescent probes has been studied to exemplify the feasibility of nanopipette-dependent targeted delivery. According to experimental results, the cytoplasm labeling of Sulfo-Cyanine5 and fluorescein isothiocyanate is mainly based on the Brownian movement due to the dyes themselves and does not have a targeting ability, while the nucleus labeling of 4′,6-diamidino-2-phenylindole (DAPI) is originated from the adsorption between DAPI and DNA in the nucleus. From the finite element simulation, the precise manipulation of intracellular delivery is realized by controlling the electro-osmotic flow inside the nanopipettes, and the different delivery modes between nontargeting dyes and nucleus-targeting dyes were compared, showcasing the valuable ability of nanopipette-based method for the analysis of specially defined subcellular regions and the potential applications for single cell surgery, subcellular manipulation, and gene delivery.
研究领域:
细胞生物学
·
分子生物学
·
生物物理学
✓ 已验证
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2021
·
IF 6.953
Fei Li,Yunlu Wei,Jing Zhao,Guoyong Yu,Linlin Huang,Quanhong Li
Pumpkin polysaccharides with various bioactivities are mainly taken orally, thus detailed knowledge of the intestinal transport of which are essential for understanding its bioactivities. The Caco-2 cells monolayer model (mimic intestinal epithelium) was successfully constructed and Cucurbia moschata polysaccharides (PPc-F) were successfully conjugated with fluorescein isothiocyanate (FITC) to evaluate the transcellular transport mechanism and subcellular localization of PPc. The transport process of PPc-F was energy-dependent, and a moderately-absorbed biological macromolecule according to the apparent permeability coefficients (P app ) value. The endocytosis process of PPc-F in Caco-2 cells included the clathrin- and caveolae (or lipid draft)-medicated routes. And the translocation process was related to endoplasmic reticulum (ER), golgi apparatus (GA), tubulin and the acidification of endosomes. As for the intracellular location of PPc-F, it was mainly accumulated in ER. The study provided an understanding of the transmembrane transport of PPc-F, and could help studying the mechanisms of its effects.
研究领域:
药理学
·
分子生物学
·
营养科学
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2017
·
IF 3.154
Lanying Wang,Yunfei Zhang,Delong Wang,Mei Wang,Yong Wang,Juntao Feng
Carabrone, a botanical bicyclic sesquiterpenic lactone, has broad-spectrum antifungal activity and is particularly efficient against the devastating phytopathogen Gaeumannomyces graminis var. tritici (Ggt). The antifungal mechanism of carabrone against Ggt, however, remains unclear. The main objective of this study was to investigate the subcellular localization of carabrone in Ggt to gain a better understanding of its mechanism of action. When Ggt was exposed to carabrone (EC50 value of 28.45 μg/mL) for 7 days, a decline in mitochondrial concentration together with some obvious alternations in mitochondrial structure, including hazy outlines, medullary transitions, excess accumulation of unclear settlings, and vacuolar degeneration, were observed, indicating that carbrone may act on the mitochondria directly. A fluorescent conjugate (TTY) was thus designed and synthesized as a surrogate of carabrone that possessed comparable antifungal activity against Ggt (EC50 of 33.68 μg/mL). Additionally, a polyclonal antibody specific to carabrone and with a high titer (256 000) was also prepared by immunizing mice. Subsequently, two imaging techniques, the use of the fluorescent conjugate (FC) and immunofluorescence (IF), were applied to determine the subcellular localization of carabrone. Both FC and IF fluorescent signals demonstrated its mitochondrial localization with a Pearson’s coefficient of 0.83 for FC and 0.86 for IF. These results imply that carabrone exerts its antifungal activity against Ggt by interfering with mitochondrial function.
研究领域:
植物病理学
·
真菌学
·
植物化学
·
细胞生物学
✓ 已验证
DOI
PubMed
高影响因子
ANALYTICAL LETTERS
·
2025
·
IF 1.8
Shancai Guo,Qizhong Hu,Xuye Zhang,Zewei Bao,Jinjun Zhao,Jianyuan Lin,Zhijian Jia,Haili Wang,Chunjie Bai,Jianghua Chen
Conventional fluorescent probes for cellular imaging face significant limitations, including photobleaching, cytotoxicity, and quenching upon aggregation (aggregation-caused quenching, ACQ). While carbon quantum dots (CQDs) offer advantages like low toxicity, their tendency toward ACQ restricts their utility. To address these challenges, novel aggregation-induced emission (AIE)-active carbon quantum dots (AIE-CQDs) were developed using the AIE principle. Synthesized hydrothermally using 2,2′-dithiobisbenzoic acid and melamine, the AIE-CQDs exhibit a unique hydrophobic structure and disulfide (S-S) bonding. Pronounced fluorescence enhancement was enabled upon aggregation due to this design. Spherical morphology (10–20 nm) and heteroatom doping (N, S) were confirmed through structural characterization. A high quantum yield (25.4%), bimodal emission (green: 484.6 nm; red: 608.8 nm), and excellent stability were exhibited by the AIE-CQDs. Minimal cytotoxicity in HeLa cells (> 90% viability at ≤ 8 μg/mL) was observed. Superior live-cell imaging was revealed by confocal laser scanning microscopy, with time-dependent intracellular aggregation, particularly near nuclear membranes, intensifying red fluorescence and achieving high-resolution imaging (0.2 μm). This AIE behavior, driven by restricted molecular motion via S-S bonding and hydrophobic interactions, resulted in an aggregated-to-dispersed fluorescence intensity ratio of approximately 3.5. AIE-CQDs were established as a promising, low-toxicity alternative with enhanced photostability and resolution for advanced cellular imaging.
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