EUROPEAN JOURNAL OF PHARMACOLOGY
·
2025
·
IF 4.7
Yi-ni Lu,Xiao-han Wang,Bo-cheng Xu,Yufen Zheng,Xiong Zhang,Yi Liu,Tian-yin Wang,Feng Yu
Septic cardiomyopathy (SCM) represents one of the life-threatening manifestations of sepsis, for which specific therapies are currently lacking. The mechanism of action of urinary trypsin inhibitor (UTI), widely employed in treating acute inflammatory disorders, on SCM has remained elusive. Here, we observed that UTI alleviated cardiac function, reduced myocardial injury, and improved cardiac permeability in a cecum ligation and puncture (CLP)-induced sepsis mouse model. In addition, UTI significantly decreased serum and myocardial angiopoietin 2 (ANG2) levels in CLP mice, which is a biomarker associated with sepsis prognosis. In vitro experiments utilizing mouse myocardial microvascular endothelial cells (MMVECs) were conducted to explore the molecular mechanisms underlying UTI-mediated regulation of ANG2 expression, secretion, and its impacts on endothelial permeability and neutrophil adhesion. UTI mitigated endothelial barrier dysfunction and reduced neutrophil adherence following thrombin stimulation and ANG2 overexpression. The experiments further demonstrated that UTI-induced inhibition of ANG2 expression was related to the blockade of forkhead box O1 (FOXO1) and phosphorylated signal transducer and activator of transcription 3 (p-STAT3) nuclear transcription and that UTI disrupted ANG2 exocytosis by reducing intracellular Ca 2+ concentration, a mechanism distinct from other serine protease inhibitors. In addition, UTI ameliorated barrier impairment and neutrophil adhesion in ANG2 overexpression cells by restoring tyrosine kinase receptor 2 Tie2)/protein kinase B (AKT) pathway phosphorylation. Docking predictions and surface plasmon resonance (SPR) further corroborated that UTI is strongly bound to ANG2. Altogether, our findings revealed a novel perspective wherein UTI functions as a multifaceted ANG2 inhibitor in SCM. Thus, ANG2 constitutes a promising therapeutic target for septic cardiomyopathy.
研究领域:
生物学
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医学
·
药理学
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细胞生物学
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DOI
PubMed
高影响因子
JOURNAL OF THE CHINESE CHEMICAL SOCIETY
·
2026
·
IF 2
Jing Chen,Xiyan He,Yuzhi Wang,Xueqin Ding
Five deep eutectic solvent (DES)-functionalized magnetic polydopamine (MPDA) composites were designed and synthesized via an ultrasound-assisted method, and subsequently integrated with magnetic solid-phase adsorption (MSPA) for the efficient adsorption of trypsin. Among them, MPDA@[ChCl][EG], functionalized with a DES composed of choline chloride and ethylene glycol, exhibited the highest adsorption efficiency. The morphology, magnetic responsiveness, and functional group distribution of MPDA@[ChCl][EG] were characterized by transmission electron microscopy, vibrating sample magnetometry, and fourier-transform infrared spectroscopy. Single-factor optimization experiments were conducted to investigate the key parameters affecting trypsin adsorption. Under optimal conditions, MPDA@[ChCl][EG] achieved an adsorption capacity of 338.99 mg·g −1 for trypsin, with an equilibrium time of 60 min. The adsorption behavior was well described by the Langmuir isotherm and pseudo-second-order kinetic models, indicating monolayer chemisorption. Furthermore, MPDA@[ChCl][EG] was successfully applied to the adsorption and separation of trypsin from a complex biological sample (bovine pancreas crude extract). This strategy of constructing magnetic adsorbents using green DESs and MPDA offers new possibilities for protein adsorption under mild conditions.
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DOI
高影响因子
JOURNAL OF BIOMATERIALS SCIENCE-POLYMER EDITION
·
2015
·
IF 1.648
Zhenqian Gu,Fangjie Wang,Haoxiang Lu,Xinling Wang,Zhen Zheng
A new kind of biodegradable poly(urethane-urea)s based on poly-lysine oligomer used as the soft segment was synthesized and characterized. In vitro degradation behavior of poly (urethane-urea)s was investigated, and was assessed by 1H NMR and mass loss. The results indicated that the peptide bonds in poly(urethane-urea)s were effectively cleaved in simulated pancreatic juice containing trypsin, while those in buffer solution without trypsin remained unaffected. The degradability was obviously improved by introducing poly-lysine oligomer into the main chain of poly(urethane-urea)s. The results of cells viability test indicated that the poly (urethane-urea)s showed a good biocompatibility on endothelial cells. The thermostability and hydrophilicity of poly(urethane-urea)s increased with increase in poly-lysine oligomer content.
研究领域:
材料科学
·
生物医学工程
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2020
·
IF 10.257
Qiong Hu,Yu Bao,Shiyu Gan,Yuwei Zhang,Dongxue Han,Li Niu
As one of the most important proteolytic enzymes , trypsin is useful as a reliable and specific biomarker for the diagnosis of pancreatitis and other pathological conditions. In this paper, a novel signal-on electrochemical biosensor based on the use of electrochemically controlled grafting of polymers as an amplification strategy is described for the ultrasensitive assay of trypsin activity. The carboxyl-group-free peptide, serving as the substrate for the recognition of trypsin, is first immobilized via its N-terminus. The tryptic cleavage of peptide substrate can generate a free carboxyl group at the C-terminus of the truncated peptide, to which through the carboxylate-Zr(IV)-carboxylate linkage the carboxyl-group-containing initiator for atom transfer radical polymerization (ATRP) can be conjugated. The subsequent surface-initiated grafting of polymers (SI-GOP) based on electrochemically controlled ATRP (eATRP), with ferrocenylmethyl methacrylate (FcMMA) as the monomer , can bring a large amount of Fc tags to electrode surface , resulting in the generation of a very high detection signal. The eATRP-based SI-GOP is easy to operate and low-cost as an amplification strategy. Under optimal conditions, the detection limit for trypsin activity can be down to 0.016 mU mL −1 (~2.68 pM or ~0.064 ng mL −1 ). As the current signal increases with trypsin activity, this trypsin biosensor is less susceptible to false positives due to the signal-on mode. Moreover, it is highly selective and applicable to inhibitor screening and the assay of trypsin activity in the presence of complex biological matrices. Taking together, this electrochemical trypsin biosensor may hold great potential in diagnostic applications.
研究领域:
分析化学
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生物化学
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生物传感器
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诊断学
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电化学
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产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
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2016
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IF 7.476
Hong Dai,Sihong Chen,Yilin Li,Baoshan Zeng,Shupei Zhang,Zhengsheng Hong,Yanyu Lin
Herein, a delicate photoelectrochemical biosensor for quantitative detection of trypsin was successfully established by virtue of polyethylenimine-sensitized TiO 2 mesocrystal as the photoactive matrix integrated with Boron-doped carbon quantum dots labeled peptide as the signal amplification tags. Specifically, polyethylenimine with fine photo-stability was introduced here as the electron transporting layer to reduce the energy barrier of TiO 2 mesocrystal, thereby facilitating the carriers transfer and improving the photocurrent response. Moreover, the Boron-doped carbon dots-peptide bioconjugates could noticeably decrease the photocurrent due to the competitively light harvesting by Boron-doped carbon dots and the steric hindrance of peptide chains, leading to less light energy arriving at the TiO 2 mesocrystal and hindering the electrons transfer between the electrolyte and electrode. The anchored conjugates synergistically promoted the decline of photocurrent signal, evidently enhancing the sensitivity of this detection protocol. When trypsin was incubated, the photoelectric signal was obviously re-promoting because arginine-containing peptide chains could be specifically cleaved by trypsin and the Boron-doped carbon quantum dots was affranchised from the electrode, making the most of the previous suppression effects released. Therefore, the intensity of photocurrent signal was proportional to the trypsin concentration in a wide linger range from 1×10 −7 mg/mL to 1.0 mg/mL. This practical and elegant “on-off-on” biosensor with high sensitivity offered a promising scheme to monitor various proteases and the inhibitors screening for early diagnoses of different diseases.
研究领域:
生物传感器技术
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分析化学
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生物医学工程
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DOI
PubMed
高影响因子
LUMINESCENCE
·
2013
·Vol. 29
·期 1
·79-86
·
IF 1.273
Li Shuai,Zhi Chen,Peng Fei,Qiang Wang,Tianming Yang
The interaction of trypsin with tetramethylpyrazine (TMP) and ferulic acid (FA) was studied using fluorescence, synchronous fluorescence, UV–vis absorption, circular dichroism (CD) and three-dimensional (3D) fluorescence spectra techniques. Using fluorescence quenching calculations, the bimolecular quenching constant ( k q ), apparent quenching constant ( K SV ), effective binding constant ( K a ) and binding site number ( n ) were obtained. The distance r between donor and acceptor was found to be 2.049 and 1.281 nm for TMP–trypsin and FA–trypsin complexes. TMP and FA can quench the fluorescence intensity of trypsin by a static quenching procedure. Thermodynamic parameters calculated on the basis of different temperatures revealed that the binding of trypsin to TMP/FA mainly depended on van der Waals' forces and hydrogen bonds. The effect of TMP and FA on the conformation of trypsin was analyzed using synchronous fluorescence, CD, 3D fluorescence spectra and molecular docking studies. Copyright © 2013 John Wiley & Sons, Ltd.
研究领域:
生物化学
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分子生物学
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光谱学
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2016
·
IF 4.712
Jie Feng,Yonglei Chen,Yangxia Han,Juanjuan Liu,Cuiling Ren,Xingguo Chen
In recent years, extensive researches are focused on the fluorescent carbon nanoparticles (CNPs) due to their excellent photochemical, biocompatible and water-soluble properties. However, these synthesis methods are generally suffered from tedious processes. In this paper, fluorescent carbon nanoparticles are synthesized by a facile, one-pot, low-temperature method with trypsin and dopamine as precursors. The synthesis process avoids any heating operation and organic solvent , which provides a “green” and effective preparation route. The obtained CNPs exhibit excellent water-solubility, salt-tolerance and photostability. Based on the synergistic action of the inner filter effect and static quenching mechanism, the CNPs are exploited as a “turn-off” fluorescence sensor for sensitive and selective detection of Fe 3+ ions . The probe shows a wide linear range from 0.1 to 500 μM, with a limit of detection of 30 nM. Furthermore, the as-fabricated fluorescent sensing system is successfully applied to the analysis of Fe 3+ in biological samples such as human urine and serum samples with satisfactory recoveries (92.8–113.3%).
研究领域:
材料科学
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分析化学
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纳米技术
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环境科学
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DOI
PubMed
高影响因子
Nanoscale Advances
·
2018
·Vol. 1
·期 1
·162-167
·
IF 0
Liping Song,Lei Zhang,Kai Xu,Youju Huang,Pan Gao,Haiyan Fang,Jiawei Zhang,Zhihong Nie,Tao Chen
Current technologies still face a big challenge to achieve simultaneous rapid qualitative and quantitative detection of trypsin. In our present study, we developed a simple and effective strategy to sensitively, qualitatively and quantitatively analyze the activity of trypsin using a fluorescent polystyrene (PS) microsphere probe. PS spheres were first functionalized by the surface coating of polyethylene glycol (PEG), which could significantly decrease the possibility of nonspecific physical adsorption of the fluorescein isothiocyanate isomer-modified peptide (peptide-FITC). Then, the obtained PS-PEG spheres were chemically interacted with peptide-FITC, which were then employed to monitor the real-time activity of trypsin. The peptide used in our work contained rich lysine and arginine residues, which were the recognition sites of trypsin. When trypsin interacted with the PS-FITC-peptide microspheres, the peptide-FITC rapidly decomposed into free small fragments in solution, resulting in a gradual decrease in the fluorescence of the PS spheres. By taking advantage of the fluorescence changes using confocal microscopy imaging and fluorescence spectrum intensity, it is easy to achieve the qualitative and quantitative detection of trypsin, with a highly sensitive detection limit as low as 0.5 ng mL−1 and high selectivity. Thus, the designed fluorescent PS microsphere probe would be very promising in various applications such as food safety inspection, personal healthcare and on-site environmental monitoring.
研究领域:
生物化学
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分析化学
·
医学检测
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DOI
PubMed
ANALYTICAL CHEMISTRY
·
2021
·
IF 6.986
Qiong Hu,Luofeng Su,Zhuohua Chen,Yanyu Huang,Dongdong Qin,Li Niu
Trypsin is a key proteolytic enzyme in the digestive system and its abnormal levels are indicative of some pancreatic diseases. Taking advantage of the coenzyme-mediated electrografting of ferrocenyl polymers as a novel strategy for signal amplification, herein, a signal-on cleavage-based electrochemical biosensor is reported for the highly selective interrogation of trypsin activity at ultralow levels. The construction of the trypsin biosensor involves (i) the immobilization of peptide substrates (without free carboxyl groups) via the N-terminus, (ii) the tryptic cleavage of peptide substrates, (iii) the site-specific labeling of the reversible addition–fragmentation chain transfer (RAFT) agents, and (iv) the grafting of ferrocenyl polymers through the electro-RAFT (eRAFT) polymerization, which is mediated by potentiostatic reduction of nicotinamide adenine dinucleotide (NAD+) coenzymes. Through the NAD+-mediated eRAFT (NAD+-eRAFT) polymerization of ferrocenylmethyl methacrylate (FcMMA), the presence of a few tryptic cleavage events can eventually result in the recruitment of a considerable amount of ferrocene redox tags. Obviously, the NAD+-eRAFT polymerization is low-cost and easy to operate as a highly efficient strategy for signal amplification. As expected, the as-constructed biosensor is highly selective and sensitive toward the signal-on interrogation of trypsin activity. Under optimal conditions, the detection limit can be as low as 18.2 μU/mL (∼72.8 pg/mL). The results also demonstrate that the as-constructed electrochemical trypsin biosensor is applicable to inhibitor screening and the interrogation of enzyme activity in the presence of complex sample matrices. Moreover, it is low-cost, less susceptible to false-positive results, and relatively easy to fabricate, thus holding great potential in diagnostic and therapeutic applications.
研究领域:
分析化学
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生物化学
·
生物技术
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电化学
·
酶学
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DOI
PubMed
高影响因子
RSC Advances
·
2017
·Vol. 7
·期 43
·26930-26934
·
IF 3.108
Wei Liu,Huanhuan Li,Yanli Wei,Chuan Dong
A label free phosphorescence sensing platform for trypsin was developed employing Mn doped ZnS quantum dots (Mn-ZnS QDs) as the phosphorescence probe. Mn-ZnS QDs can emit strong room temperature phosphorescence. Upon addition of Cytochrome c (Cyt c), a hybrid complex between Mn-ZnS QDs and Cyt c was formed, and consequently, quenched the phosphorescence of Mn-ZnS QDs. In the presence of trypsin, Cyt c was hydrolyzed into small fragments, and accordingly, the quenched phosphorescence of Mn-ZnS QDs went back and high phosphorescence intensity was observed. As such, based on the phosphorescence signal changes of the system, a label free turn-on phosphorescence sensing platform was fabricated. Factors affecting the determination of trypsin including the concentration of Mn-ZnS QDs, reaction time, temperature and pH effect were comprehensively explored. Under the optimal experimental conditions the restored phosphorescence of the sensing system displayed a response for trypsin in the range of 0.88–15.6 μg mL−1 with a detection limit of 42 ng mL−1. Additionally, the sensing platform was successfully applied for the determination of trypsin in human serum samples with satisfactory recovery.
研究领域:
分析化学
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生物传感器
·
纳米技术
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DOI
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2013
·
IF 2.906
Dong-Fang Li,Hao-Chen Ding,Tao Zhou
A commercially available trypsin–chymotrypsin mixture was covalently immobilized onto modified polyvinyl chloride (PVC) microspheres, which were activated by the subsequent treatment of PVC microspheres with ethylenediamine and glutaraldehyde. The immobilized mixed protease was characterized by FT-IR and SEM analyses. Immobilization conditions were optimized by Box–Behnken design and the response surface method. The activity of the immobilized mixed protease prepared under optimal conditions (pH 6.6, 23 °C, 2 h) reached 1341 U/g. Compared with the free form, the immobilized enzyme possesses a slightly higher optimal pH value and a wider pH-activity profile, superior thermal stability, and a higher Km value. Reusability of the immobilized mixed protease indicated that >70% of the original activity was retained after having been recycled six times.
研究领域:
生物化学
·
酶工程
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DOI
PubMed
高影响因子
JOURNAL OF LUMINESCENCE
·
2013
·
IF 2.144
Ruiyong Wang,Xiaohui Kang,Ruiqiang Wang,Rui Wang,Huanjing Dou,Jing Wu,Chuanjun Song,Junbiao Chang
The interactions between trypsin and caffeine/theophylline were investigated by fluorescence spectroscopy , UV–visible absorption spectroscopy , resonance light scattering and synchronous fluorescence spectroscopy under mimic physiological conditions . The results revealed that the fluorescence quenching of trypsin by caffeine and theophylline was the result of the formed complex of caffeine–trypsin and theophylline–trypsin. The binding constants and thermodynamic parameters at three different temperatures were obtained. The hydrophobic interaction was the predominant intermolecular forces to stabilize the complex. Results showed that caffeine was the stronger quencher and bound to trypsin with higher affinity than theophylline .
研究领域:
生物化学
·
分子药理学
·
药物代谢
·
蛋白质-配体相互作用
·
光谱学
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DOI
高影响因子
危险品化学品经营许可证（带存储）