Exogenous Cys Research Articles

Novel Colorimetric and Near-Infrared Ratiometric Fluorescent Probe for Sensing Cysteine in Food Samples, Plants, and Living Cells.

Cysteine (Cys) is a crucial biothiol that acts a significant function in food samples and biological systems, including plant roots and living cells. Hence, we developed a novel colorimetric and near-infrared ratiometric fluorescent probe (CT), composed of coumarin and tetrahydroacridine-conjugated indole salt, for the detection of Cys. Upon reaction with Cys, the probe undergoes a specific N-substitution reaction, resulting in a notable colorimetric change and a significant ratiometric fluorescent response in both visible and near-infrared emission channels. These dual-channel ratiometric fluorescence changes are completely independent, enabling the probe to obtain great selectivity, sensitivity, and exceptional detection accuracy. Leveraging these attributes, the probe was employed to provide accurate quantitative analysis of Cys in food samples. Furthermore, confocal imaging demonstrated that the probe could monitor both exogenous and endogenous Cys levels in living cells and track Cys changes in plant roots under heavy metal stress. This work presents a dependable and accurate imaging solution for tracking and identifying Cys of real food, plants, and living cells.

Preparation and characterization of cysteine-rich collagen peptide and its antagonistic effect on microplastic induced damage to HK-2 cells

Cysteine (Cys) plays a significant role in the stability and antioxidant capacity of peptide. This study was to prepare Cys-rich collagen peptide and evaluate its preventive effect on microparticle (MPs)-induced HK-2 cell damage. The results showed that the Cys-rich peptides (AMP-C) were successfully prepared from collagen hydrolysate by plastein reaction in the presence of exogenous Cys. The scavenging activities of 2,2-Diphenyl-1-picrylhydrazyl and hydroxyl radical of AMP-C were significantly higher than those of original peptides without Cys. The fraction with molecular weight < 3 kDa (AMP-CA) significantly improved MPs-induced damage and apoptosis of HK-2 cells. Compared with the same fractions lacking Cys (AMP-A), AMP-CA showed better performance in reducing reactive oxygen species and increasing the total superoxide dismutase and glutathione peroxidase levels, which may be related to its down-regulation of Kelch-like ECH-associated protein 1 (Keap1) expression. In addition, AMP-CA showed strong effects on inhibiting interleukin-6, tumor necrosis factor-α, monocyte chemoattractant protein-1, kidney injury molecule-1, and nuclear factor kappa-B levels. Finally, 330 Cys-containing peptides were isolated from AMP-CA, among which Leucine-Glycine-Asparagine-Glycine-Cysteine-Proline (LGNGCP) showed the lowest binding energy with Keap1 protein. In conclusion, the Cys-rich collagen peptide with higher antioxidant activity was successfully prepared by plastein reaction, which alleviated the HK-2 cells damage induced by MPs through antioxidant and anti-inflammatory. Our findings suggested that Cys-rich collagen peptide might be used as a functional food to mitigate health hazards associated with MPs accumulation.

Dual-Channel fluorescent detection of Biothiols: A novel probe for Distinguishing Cysteine, Homocysteine, Glutathione, and N-Acetylcysteine in cellular environments

Biothiols, including cysteine (Cys), homocysteine (Hcy), glutathione (GSH), and N-acetylcysteine (NAC), possess similar chemical structures and properties but play crucial, distinct roles in biological cells and blood serum. Imbalances in the concentrations of these biothiols are associated with various diseases, highlighting the importance of precise discrimination, especially between Cys and other biothiols. Owing to the similarity of the chemical properties of Cys, Hcy, GSH, and NAC, developing an effective methodology to differentiate these thiol compounds is challenging. In this study, we designed and synthesized a novel dual-channel fluorescent probe, hereafter referred to as CNTC, by integrating coumarin and acrylonitrile. This probe enabled the simultaneous discrimination of Cys from Hcy, GSH, and NAC, producing distinct fluorescent signals: blue for Cys and green for Hcy, GSH, and NAC. CNTC exhibited rapid response kinetics (within 30 min) and impressive detection limits of 0.31, 0.11, 0.029, and 0.032 μM for Cys, Hcy, GSH, and NAC, respectively. Furthermore, CNTC was successfully applied in the fluorescence imaging of both exogenous and endogenous Cys, Hcy, GSH, and NAC in living cells. The remarkable analytical and bioimaging capabilities of CNTCin vivo establish it as a promising tool for elucidating the pathophysiological roles of biothiols, particularly Cys, Hcy, GSH, and NAC.

A responsive aggregation-induced emission fluorescent probe for the detection of cysteine in food, serum samples and oxidative stress environments

Heavy metal ions tend to accumulate through the food chain and may cause many serious diseases. As an antioxidant, cysteine (Cys) plays a crucial role in regulating redox homeostasis. In addition, Cys is also widely used in the food industry. Therefore, the development of a convenient, sensitive, and practical tool for detecting Cys is of great significance for investigating the physiological and pathological functions of Cys induced by heavy metal ions. In this paper, a fluorescent probe NYVB with aggregation-induced emission (AIE) activities has been designed and synthesized, which featured good advantages in Cys detection, including favorable selectivity, desirable sensitivity, wide linear range, and low detection limit. The prepared solid-state sensor enabled the ratiometric visual detection of Cys. Probe NYVB has been successfully implemented for testing Cys in real food samples and serum samples with satisfactory recoveries. In addition, probe NYVB has been successfully employed for detecting endogenous and exogenous Cys in SKOV3 cells. More excitingly, probe NYVB has also been used to track the detection of Cys in H2O2, Hg2+, or Cu2+-induced redox imbalance. Overall, probe NYVB will open up a new avenue for studying Cys in the amelioration of heavy metal toxicity and has practical applications.

Exogenous Application of Amino Acids Alleviates Toxicity in Two Chinese Cabbage Cultivars by Modulating Cadmium Distribution and Reducing Its Translocation.

Plants communicate underground by secreting multiple amino acids (AAs) through their roots, triggering defense mechanisms against cadmium (Cd) stress. However, the specific roles of the individual AAs in Cd translocation and detoxification remain unclear. This study investigated how exogenous AAs influence Cd movement from the roots to the shoots in Cd-resistant and Cd-sensitive Chinese cabbage cultivars (Jingcui 60 and 16-7 cultivars). The results showed that methionine (Met) and cysteine (Cys) reduced Cd concentrations in the shoots of Jingcui 60 by approximately 44% and 52%, and in 16-7 by approximately 43% and 32%, respectively, compared to plants treated with Cd alone. However, threonine (Thr) and aspartic acid (Asp) did not show similar effects. Subcellular Cd distribution analysis revealed that AA supplementation increased Cd uptake in the roots, with Jingcui 60 preferentially storing more Cd in the cell wall, whereas the 16-7 cultivar exhibited higher Cd concentrations in the organelles. Moreover, Met and Cys promoted the formation of Cd-phosphate in the roots of Jingcui 60 and Cd-oxalate in the 16-7 cultivar, respectively. Further analysis showed that exogenous Cys inhibited Cd transport to the xylem by downregulating the expression of HMA2 in the roots of both cultivars, and HMA4 in the 16-7 cultivar. These findings provide insights into the influence of exogenous AAs on Cd partitioning and detoxification in Chinese cabbage plants.

Visualizing detection of cysteine level under LPS-induced oxidative stress process using benzopyrylium-based mitochondrial-targeted NIR fluorescent probe

As an important scavenger of reactive oxygen species, cysteine (Cys) plays a crucial role in mitochondrial oxidative stress. Abnormal intracellular Cys concentration under oxidative stress leads to various diseases. Cys is also widely used in the food industry. Many vegetables and fruits are rich in Cys. Abnormal levels of Cys in the body can lead to a variety of diseases. Therefore, the design of fluorescent probe that can detect Cys quickly is particularly important in living organisms. In this work, a novel turn-on mitochondrial-targeted NIR fluorescent probe (SXNU-6) was designed for the high sensitivity and specificity of Cys based on benzopyrylium. Among them, acrylate group acted as recognition site and fluorescence quencher. With the addition of Cys, the fluorescence of SXNU-6 was gradually enhanced at 660 nm. The probe had good selectivity and high sensitivity, which could realize the detection of Cys in food samples (onion, garlic, tomato, lettuce, apple, pear, milk and bread). Also, SXNU-6 could detect both endogenous and exogenous Cys under LPS-induced oxidative stress process. This work provide a powerful tool for detecting Cys in food samples and biological systems.

A two-photon fluorescent probe for highly selective detection of Cys over GSH and Hcy based on the Michael addition and transcyclization mechanism and its application in bioimaging and protein straining in SDS-PAGE

BackgroundCysteine (Cys), glutathione (GSH), and homocysteine (Hcy), as three major biothiols are involved in a variety of physiological processes and play a crucial role in plant growth. Abnormal levels of Cys can cause plants to fail to grow properly. To date, although a very large number of fluorescent probes have been reported for the detection of biothiols, very few of them can be used for the selective discrimination of Cys from GSH and Hcy due to their structural similarity, and only a few of them can be used for plant imaging. ResultsHere, three fluorescent probes (o-/m-/p-TMA) based on TMN fluorophore and the ortho-/meta-/para-substituted maleimide recognition groups were constructed to investigate the selective response effect of Cys. Compared to the o-/m-TMA, p-TMA can selectively detect Cys over GSH and Hcy with a rapid response time (10 min) and a low detection limit (0.26 μM). The theoretical calculation confirmed that the intermediate p-TMA-Cys-int has shorter interatomic reaction distances (3.827 Å) compared to o-/m-TMA-Cys (5.533/5.287 Å), making it more suitable for further transcyclization reactions. Additionally, p-TMA has been employed for selective tracking of exogenous and endogenous Cys in Arabidopsis thaliana using both single-/two-photon fluorescence imaging. Furthermore, single cell walls produced obvious two-photon fluorescence signals, indicating that p-TMA can be used for high-concentration Cys analysis in single cells. Surprisingly, p-TMA can be used as a fluorescent dye for protein staining in SDS-PAGE with higher sensitivity (7.49 μg/mL) than classical Coomassie brilliant blue (14.11 μg/mL). SignificanceThe outstanding properties of p-TMA make it a promising multifunctional molecular tool for the highly selective detection of Cys over GSH and Hcy in various complex environments, including water solutions, zebrafish, and plants. Additionally, it has the potential to be developed as a fluorescent dye for a simple and fast SDS-PAGE fluorescence staining method.

Lysosomal-targeted fluorescent probe based pH regulating reactivity for tracking cysteine dynamics under oxidative stress.

The ability to detect and visualize cellular events and associated biological analytes is essential for the understanding of their physiological and pathological functions. Cysteine (Cys) plays a crucial role in biological systems and lysosomal homeostasis. This puts forward higher requirements on the performance of the probe. Herein, we rationally designed a coumarin-based probe for the reversible, specific, sensitive, and rapid detection of Cys based on pH regulating reactivity. The obtained probe (ECMA) introduces a morpholine moiety to target lysosomes, and α,β-unsaturated-ketone with an electron-withdrawing CN group served as a reversible reaction site for Cys. Importantly, ECMA was successfully applied to the real-time monitoring of Cys dynamics in living cells. Furthermore, cell imaging clearly revealed that exogenous Cys could induce the up-regulation of lysosomal ROS, which provided a powerful tool for investigating the relationship between oxidative stress and lysosomal Cys.

High-selective two-site fluorescent probe for Cys/SO2 detection and cell imaging

A fluorescent probe has been designed using cyanine phenothiazine and 7-nitro-1,2,3-benzoxadiazole (NBD) for selective detection of Cys-SO2 components. The probe utilizes the NBD structure to achieve specificity towards Cys and employs a reaction mechanism between the double bond of cyanine and phenothiazine with SO32− to achieve selectivity towards SO2. Importantly, the NBPI phenothiazine structure incorporates a large C-O bond energy attached to NBD, effectively eliminating interference from Hcy and ensuring highly selective response to Cys. The optimized design of the probe enables excellent linearity and extremely low detection limits for Cys-SO2 components in vitro experiments. The probe NBPI allows separate detection of Cys-SO2 in the presence of both components. Furthermore, the probe NBPI demonstrated successful imaging of endogenous and exogenous Cys and SO2 in cell studies.

Near-infrared colorimetric and ratiometric fluorescent probe for dual-channel imaging of mitochondria and cysteine in the oxidative stress model

Mitochondria, known as the “cellular powerhouse”, play a variety of fundamental roles in various physiological and pathological processes, whose oxidative damage is usually emerging as a crucial induction factor that not only induces apoptosis but may also contribute to epilepsy and some other related diseases. Therefore, it is of notable significance to construct a high-discrimination method for informing the diagnosis and treatment of related diseases. Employing Cys-triggered conjugate addition cyclization-elimination strategy, we rationally constructed a mitochondria-targeted colorimetric and ratiometric near-infrared (NIR) fluorescent probe, named Mito-Pa, for Cys-triggered dual-channel fluorescence imaging of mitochondrial oxidative damage during apoptosis. Taking advantage of good membrane permeability and low cytotoxicity, probe Mito-Pa was successfully applied to visualize the mitochondria during apoptotic and the fluctuation of endogenous and exogenous Cys, or generated upon lipopolysaccharide-induced oxidative stress, in various living systems, including cancer cells, zebrafish, and mice. Thus, our proposed probe Mito-Pa would provide fresh insight for a better understanding of the ultimate relationship of Cys with mitochondrial oxidative damage, enabling easy access to accurately monitor the occurrence and progress of related diseases.

An extra-large Stokes shift near-infrared fluorescent probe for specific detection and imaging of cysteine

Cysteine (Cys) plays a crucial role in numerous physiological and pathological processes. Therefore, it is imperative to design a highly selective and sensitive near-infrared (NIR) fluorescent probe to monitor Cys. In this study, we have developed a novel NIR fluorescent probe XA based on Xanthene hybrid tetrahydro-acridine salt dye for specifically tracking of Cys, where a chlorine-substituted tetrahydro-acridine acts as a high Cys-reactive site and water-soluble group. Probe XA exhibits a remarkable "turn-on" NIR emission (830 nm) with an extra-large Stokes shift (305 nm) for monitoring Cys. It also has a high selectivity, rapid response time (6 min) and high sensitivity (LOD as 0.5 μM). We fully characterized and discussed the sensing mechanism of XA toward Cys using HPLC and MS spectrums, as well as quantum theory calculations. Furthermore, the excellent properties of NIR fluorescent detection allow this novel probe to successfully monitor fluctuations of exogenous and endogenous Cys concentration levels in living cells and in vivo.

Real-time monitoring of endogenous cysteine in LPS-induced oxidative stress process with a novel lysosome-targeted fluorescent probe

Cysteine (Cys), one of essential small-molecule-based biothiols in the human body, contributes to the regulation of redox reactions and is closely associated with many physiological and pathological metabolic processes. Herein, a novel fluorescent probe, hydroxyphenyl-conjugated benzothiazole (HBT-Cys) capable of detecting Cys was constructed, where acrylate served as the recognition group and hydroxyphenyl-linked benzothiazole acted as the fluorophore. The fluorescence of the probe was negligible in the absence of Cys, and an intense blue fluorescence was observed upon addition of Cys. The Cys-sensing mechanism could be ascribed to the Cys-involved hydrolysis reaction with acrylate, leading to light up the emission at 430 nm with about 80-fold enhancement. In addition, HBT-Cys exhibited a fast response time, remarkable selectivity and low detection limit. HBT-Cys also worked well in real-time monitoring of Cys in three different food samples (wolfberry, hawthorn, and red dates). Importantly, our probe had an excellent lysosomes-targeted ability, which was successfully employed to real-time visualize the fluctuation of both exogenous and endogenous Cys in living cells and zebrafish under lipopolysaccharide (LPS)-induced oxidative stress. Hopefully, the work shown here provides a potent candidate for the real-time tracking of Cys fluctuations in various biological samples.

A new lysosome-targeted Cys probe and its application in biology and food samples

Cysteine (Cys) is a sulfur-containing amino acid that plays an important role in living systems. The most common way to supplement the body with exogenous Cys is through the consumption of Cys-rich foods. Therefore, it is important to detect and analyze Cys in living systems and food samples. However, most of the Cys fluorescent probes developed so far are limited to the detection of the cellular environment only, and very few probes can take into account the detection of Cys in plant roots and food samples. In this paper, a novel fluorescent probe LN-NCS targeting the detection of Cys in lysosomes was designed and synthesized by modifying the naphthalimide fluorophore. The probe LN-NCS has a large Stokes shift (140 nm), low cytotoxicity, low detection limit (16.3 nM), and high selectivity, and probe LN-NCS reacts with Cys to produce the compound LN-NH2 with good fluorescence quantum yield (Ф = 0.81). Probe LN-NCS can be used to detect Cys in cells, zebrafish, plant roots, food samples, and environmental water samples. In addition, by modeling cellular inflammation, we have demonstrated that probe LN-NCS can detect changes in Cys concentration induced by cellular inflammation, providing a potential tool to better study the cellular inflammatory environment.

A New Lysosome-Targeted NIR Fluorescent Probe for Specific Detection of Cysteine over Homocysteine and Glutathione.

In this paper, by modifying the thioxanthene-benzothiozolium fluorophore, BCy-Cys, a lysosome-targeted near-infrared (NIR) fluorescent probe was synthesized for the detection of cysteine (Cys) from homocysteine (Hcy)/glutathione (GSH). As expected, BCy-Cys exhibited high selectivity and high sensitivity for detection of Cys over Hcy/GSH, with an extremely low limit of detection at 0.31 μM, marked by obvious color changes. HRMS was conducted to confirm that the fluorescence intensity at 795 nm was significantly enhanced by the enhancement of intramolecular charge transfer (ICT). Importantly, BCy-Cys could be used to visualize both exogenous and endogenous lysosomal Cys, signifying its potential application in complex organismal systems.

Transcriptomic analysis reveals the functions of H2S as a gasotransmitter independently of Cys in Arabidopsis.

Numerous studies have revealed the gasotransmitter functions of hydrogen sulfide (H2S) in various biological processes. However, the involvement of H2S in sulfur metabolism and/or Cys synthesis makes its role as a signaling molecule ambiguous. The generation of endogenous H2S in plants is closely related to the metabolism of Cys, which play roles in a variety of signaling pathway occurring in various cellular processes. Here, we found that exogenous H2S fumigation and Cys treatment modulated the production rate and content of endogenous H2S and Cys to various degrees. Furthermore, we provided comprehensive transcriptomic analysis to support the gasotransmitter role of H2S besides as a substrate for Cys synthesis. Comparison of the differentially expressed genes (DEGs) between H2S and Cys treated seedlings indicated that H2S fumigation and Cys treatment caused different influences on gene profiles during seedlings development. A total of 261 genes were identified to respond to H2S fumigation, among which 72 genes were co-regulated by Cys treatment. GO and KEGG enrichment analysis of the 189 genes, H2S but not Cys regulated DEGs, indicated that these genes mainly involved in plant hormone signal transduction, plant-pathogen interaction, phenylpropanoid biosynthesis, and MAPK signaling pathway. Most of these genes encoded proteins having DNA binding and transcription factor activities that play roles in a variety of plant developmental and environmental responses. Many stress-responsive genes and some Ca2+ signal associated genes were also included. Consequently, H2S regulated gene expression through its role as a gasotransmitter, rather than just as a substrate for Cys biogenesis, and these 189 genes were far more likely to function in H2S signal transduction independently of Cys. Our data will provide insights for revealing and enriching H2S signaling networks.

Novel V-shaped D-π-A-π-D two-photon absorption compounds with large Stokes shifts: Synthesis, optical properties, selective detection of cysteine, and imaging in living cells

The novel V-shaped donor-π-acceptor-π-donor compounds bearing an aldehyde group (1Pip, 2EtO, and 3TEG) were designed, synthesized, and evaluated as the two-photon excited fluorescence probes for the selective detection of cysteine (Cys). Their structures were fully characterized by 1H NMR, 13C NMR, FT-IR, and HRMS. The optical properties of the target compounds in various solvents were studied in details and the time-dependent density functional theory calculations were performed to shed light on their electronic structures. It is found that all of them exhibit large Stokes shifts (6331-10047 cm−1). 1Pip and 3TEG are capable of detecting Cys with high selectivity, good anti-interference ability, and favorable two-photon absorption performance. The limits of detection were determined to be equal to 0.15 μM for 1Pip and 0.73 μM for 3TEG. The possible recognition mechanism was also validated by 1H NMR and HRMS. One- and two-photon fluorescence imaging experiments indicate that 3TEG shows more satisfactory cellular membrane permeability than 1Pip, and it can successfully visualize the endogenous and exogenous Cys in living cells. These results imply that 3TEG has potential as a two-photon fluorescence probe for detecting Cys in biological samples.

Photoacoustic/fluorescence dual-modality cyanine-based probe for real-time imaging of endogenous cysteine and in situ diagnosis of cervical cancer in vivo

Cysteine (Cys), one of cellular biothiols in the organism, is associated with many diseases, such as Parkinson's disease, Alzheimer's disease, liver damage, rheumatoid arthritis, and cancer. However, activatable fluorescence/photoacoustic (FL/PA) probes for non-invasive, real-time, and deep imaging of Cys in vivo are still lacking, and this hinders the diagnosis of Cys-related diseases. Herein, we designed and synthesized a novel activated FL/PA dual-modality cyanine-base probe (FP700) for real-time detection of Cys. The probe FP700 was established with near-infrared emissive dye cyanin as the fluorophore, linking with 2, 4-dinitrobenzenesulfonyl group as the recognition moiety for Cys. Using the FP700, we found that the FP700 exhibited intensive “turn-on” FL/PA signals under the excitation of 700 nm, which realized noninvasive in vivo detection of exogenous Cys. Significantly, FP700 accurately detected endogenous Cys though the FL/PA dual-mode imaging technology in tumor-bearing mice and obtain 3D PA diagnostic images with deep penetration depth and spatial resolution. Thus, the new dual-modality probe FP700 has advantage of high potential for deep tumor diagnosis of Cys in vivo, which may provide a new approach for the detection of cervical cancer and identification of its potential therapeutic targets.

Competitive Formation of 2,3-Butanedione and Pyrazines through Intervention of Added Cysteine during Thermal Processing of Alanine-Xylose Amadori Compounds.

The intervention of cysteine (Cys) on the formation of 2,3-butanedione and pyrazines was evaluated during the thermal processing of the alanine-xylose Amadori compound (AX-ARP). With the involvement of Cys, the competitive formation of 2,3-butanedione and pyrazines was induced. The formation of 2,3-butanedione in the AX-ARP/Cys model was suppressed due to the inhibitory effect of the precursors of 2,3-butanedione like deoxypentosones, while the added Cys in the AX-ARP/Cys model competed with the recovered alanine (Ala) to capture glyoxal and methylglyoxal to make up for the absence of pyrazines in the AX-ARP model at an initial pH value of 7. The content of pyrazines increased from 0 up to 16.48 μg/L (120 °C, 120 min). Exogenous Cys itself showed lower reactivity with 2,3-butanedione through the Strecker degradation reaction; while the pH was increased to 8, the degradative products of Cys were facilitated to consume the residual 2,3-butanedione giving rise to the formation of 2,4,5-trimethylthiazole at 120 °C. It was the degradative products of Cys that accelerated the reaction for consumption of 2,3-butanedione rather than Cys itself. Additionally, the inhibitory effect of Cys on 2,3-butanedione formation was weakened under a basic condition, while the promotional effect on the formation of pyrazines was further boosted. With more Cys participating in the process of AX-ARP thermal degradation, the formation of 2,3-butanedione was further inhibited, while the yields of pyrazines were increased.

A novel flavonol-based fluorescent probe for rapid detection of Cysteine in food samples and its applications in bioimaging systems

l-Cysteine (Cys) plays an important role in the food industry and the physiological processes. Herein, a new fluorescent turn-on probe 6-(4-(Diphenylamino)phenyl)-2-(furan-2-yl)-3-hydroxychromone acrylate (DDPF) with AIE property based on flavonol was developed for rapidly detecting Cys. This probe displayed a significantly enhanced fluorescence response to Cys over other various competing analytes such as amino acids, cations, and anions. Probe DDPF possesses some good advantages in the detection of Cys including large Stokes shift (195 nm), low detection limit (97.6 nM), wide pH range (3–8), and fast response time (50 s). In addition, DDPF was also employed for the quantitative determination of trace Cysteine in dairy product samples, such as milk, milk powder, cheese, and yogurt. Moreover, this probe had good biocompatibility and could visualize endogenous and exogenous Cys in the living cells. Furthermore, it could be successfully applied for fluorescence imaging Cys in zebrafish.

Cysteine-activated fluorescence/photoacoustic integrated probe for non-invasive diagnosis of drug-induced liver injury

Hepatotoxicity is a serious problem faced by clinical drugs, and long-term administration or overdose may lead to liver failure and even death of patients. Therefore, developing a reliable detection method for the early diagnosis and therapy of drug-induced liver injury (DILI) has significant meaning. Near-infrared fluorescence (NIRF) and photoacoustic (PA) dual-modality tomography probes can be used for imaging with high sensitivity and high-resolution of disease-related markers in deep tissues. Here, we developed a novel Cys-activated NIRF and PA dual-modality imaging probe (CDR) for early diagnosis of DILI, for the first time. The organic molecular probe CDR could respond rapidly to Cys, resulting in the absorption peak red-shifted from 560 nm to 725 nm, which also leads to the activation of the PA725 signal and NIRF765 signal. In addition, the new probe CDR could be used for NIRF/PA imaging of exogenous and endogenous Cys level in live cells and mice. More importantly, CDR has also been successfully used for in situ detection of Cys in early DILI mice and evaluate the therapeutic effect of NAC. Therefore, the CDR might become a powerful tool to research the physiological effect of Cys and evaluate the degree of DILI.