ACS Applied Materials & Interfaces
·
2024
·
IF 8.3
Shengjie Liu,He Yang,Xingyu Heng,Lihua Yao,Wei Sun,Qing Zheng,Zhaoqiang Wu,Hong Chen
To effectively solve the problem of significant loss of transplanted cells caused by thrombosis during cell transplantation, this study simulates the human fibrinolytic system and combines metabolic oligosaccharide engineering with strain-promoted azide–alkyne cycloaddition (SPAAC) click chemistry to construct a cell surface with fibrinolytic activity. First, a copolymer (POL) of oligoethylene glycol methacrylate (OEGMA) and 6-amino-2-(2-methylamido)hexanoic acid (Lys) was synthesized by reversible addition–fragmentation chain transfer (RAFT) copolymerization, and the dibenzocyclooctyne (DBCO) functional group was introduced into the side chain of the copolymer through an active ester reaction, resulting in a functionalized copolymer DBCO-PEG4-POL with ε-lysine ligands. Then, azide functional groups were introduced onto the surface of HeLa model cells through metabolic oligosaccharide engineering, and DBCO-PEG4-POL was further specifically modified onto the surface of HeLa cells via the SPAAC “click” reaction. In vitro investigations revealed that compared with unmodified HeLa cells, modified cells not only resist the adsorption of nonspecific proteins such as fibrinogen and human serum albumin but also selectively bind to plasminogen in plasma while maintaining good cell viability and proliferative activity. More importantly, upon the activation of adsorbed plasminogen into plasmin, the modified cells exhibited remarkable fibrinolytic activity and were capable of promptly dissolving the primary thrombus formed on their surfaces. This research not only provides a novel approach for constructing transplantable cells with fibrinolytic activity but also offers a new perspective for effectively addressing the significant loss of transplanted cells caused by thrombosis.
研究领域:
生物化学
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细胞生物学
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材料科学
·
药学
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生物医学工程
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DOI
PubMed
高影响因子
Polymer Chemistry
·
2013
·Vol. 4
·期 22
·5597-5602
·
IF 5.231
Zengchao Tang,Xiaoli Liu,Yafei Luan,Wei Liu,Zhaoqiang Wu,Dan Li,Hong Chen
A new strategy to regulate the attachment of plasminogen and its activator, t-PA, to a polyurethane surface is reported. The method is based on the one-step graft copolymerization of a lysine-containing methacrylic monomer and hydroxyethyl methacrylate (HEMA). The copolymer-modified PU surfaces showed high capacity for plasminogen and t-PA binding. The release of pre-adsorbed t-PA from the lysine-containing PU surfaces in contact with plasma was also investigated. It was shown that the interactions of plasminogen and t-PA on this type of surface can be controlled simply by changing the monomer feed ratio, and thus the relative quantities of HEMA and lysine in the grafts. This approach may be useful for the development of fibrinolytic, blood-contacting materials and, more generally, for controlling protein-surface interactions for biocompatibility.
研究领域:
生物材料学
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医学工程
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DOI
高影响因子
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
·
2023
·
IF 3.1
Chunying Guo,Ting Wang,Dongmei Zhang,Xiaojing Ge,Jinan Li
Alzheimer's disease (AD) is the most common neurodegenerative disorder in the world. The aggregation of both amyloid beta (Aβ) peptides extracellularly and Tau proteins intracellularly plays key roles in the pathological consequences of AD, which lead to cholinergic neurodegeneration and eventually death. Currently, there are no effective methods to stop the progression of AD. Using ex vivo , in vivo and clinical approaches, we investigated the functional effects of plasminogen on the widely used FAD, Aβ42 oligomer or Tau intracranial injection-induced AD mouse model and explored its therapeutic effects on patients with AD. The results show that intravenously injected plasminogen rapidly crosses the blood‒brain barrier (BBB); increases plasmin activity in the brain; colocalizes with and effectively promotes the clearance of Aβ42 peptide and Tau protein deposits ex vivo and in vivo ; increases the choline acetyltransferase (ChAT) level and decreases the acetylcholinesterase (AChE) activity; and improves the memory functions. Clinically, when GMP-level plasminogen was administered to 6 AD patients for 1–2 weeks, their average scores on the Minimum Mental State Examination (MMSE), which is a standard scoring system used to measure the memory loss and cognitive deficits, were extremely significantly improved by 4.2 ± 2.23 points, e.g., an average increase from 15.5 ± 8.22 before treatment to 19.7 ± 7.09 after treatment. The preclinical study and pilot clinical study suggest that plasminogen is effective in treating AD and may be a promising drug candidate.
研究领域:
神经科学
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药理学
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临床试验
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DOI
PubMed
高影响因子
ANALYST
·
2017
·Vol. 142
·期 24
·4773-4781
·
IF 3.885
Haijiao Zheng,Naizhong Song,Xiqian Li,Qiong Jia
Plasminogen (Plg) is a kind of glycoprotein which plays an important role in cell migration. The determination of Plg content can directly reflect the abnormal manifestation of fibrinolytic system dysfunctions. In the present work, lysine (Lys)-based adsorbents were prepared for the specific capture of Plg through the covalent binding of Lys with a polymer monolithic substrate. Lys was modified with β-cyclodextrin (β-CD) via a click reaction and anchored to the substrate with biotin by host–guest interaction. The biotin-Lys-CD based monolithic material was employed for the specific capture of Plg. Combining with mass spectrometry (MS) determinations, the method exhibited a low detection limit of 1.0 fmol with relative standard deviations below 10.0% for Plg. Considering the advantages of simplicity, sensitivity, and high specificity, the developed approach was successfully applied to the determination of Plg in human plasma samples and opened a gallery for testing Plg as a biomarker for the diagnosis of fibrinolytic system dysfunctions.
研究领域:
生物化学
·
医学诊断
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DOI
PubMed
高影响因子
Marine Drugs
·
2024
·Vol. 22
·期 2
·68
·
IF 4.9
Tingting Jiang,Bing Zhang,Haixing Zhang,Mingjun Wei,Yue Su,Tuo Song,Shijia Ye,Yuping Zhu,Wenhui Wu
Marine organisms are a rich source of enzymes that exhibit excellent biological activity and a wide range of applications. However, there has been limited research on the proteases found in marine mudflat organisms. Based on this background, the marine fibrinolytic enzyme FELP, which was isolated and purified from clamworm (Perinereis aibuhitensis), has exhibited excellent fibrinolytic activity. We demonstrated the FELP with a purification of 10.61-fold by precipitation with ammonium sulfate, ion-exchange chromatography, and gel-filtration chromatography. SDS-PAGE, fibrin plate method, and LC–MS/MS indicated that the molecular weight of FELP is 28.9 kDa and identified FELP as a fibrinolytic enzyme-like protease. FELP displayed the maximum fibrinolytic activity at pH 9 (407 ± 16 mm2) and 50 °C (724 ± 27 mm2) and had excellent stability at pH 7–11 (50%) or 30–60 °C (60%), respectively. The three-dimensional structure of some amino acid residues of FELP was predicted with the SWISS-MODEL. The fibrinolytic and fibrinogenolytic assays showed that the enzyme possessed direct fibrinolytic activity and indirect fibrinolysis via the activation of plasminogen; it could preferentially degrade Aα-chains of fibrinogen, followed by Bβ- and γ-chains. Overall, the fibrinolytic enzyme was successfully purified fromPerinereis aibuhitensis,a marine Annelida (phylum), with favorable stability that has strong fibrinolysis activity in vitro. Therefore, FELP appears to be a potent fibrinolytic enzyme with an application that deserves further investigation.Keywords:Perinereis aibuhitensis;plasmin-like protease;amino acid residue sequences;fibrinolysis in vitro
研究领域:
海洋生物学
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酶学
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生物化学
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DOI
PubMed
高影响因子
Food Chemistry-X
·
2023
·
IF 6.1
Jialin Wu,Guangqun Lan,Na He,Laping He,Cuiqin Li,Xiao Wang,Xuefeng Zeng
A producing-fibrinolytic enzyme strain was isolated with high yield. The strain was identified as Bacillus amyloliquefaciens . B. amyloliquefaciens GUTU06 fibrinolytic enzyme was purified by acetone precipitation and reverse micelle. Acetone precipitation condition and reverse micelle condition were examined. Results showed that the total reverse micelle extraction efficiency was 64.49 % ± 1.6 %. The purification fold of the entire process reached 13.38. The optimum pH of purified enzyme is 5, and the optimum temperature is 45 °C. Fe 3+ and K + can enhance the fibrinolytic activity of the enzyme. Compared to commercial fibrinolytic enzymes such as urokinase and lumbrukinase, GUTU06 fibrinolytic enzymes have a lower pH optimal range and higher temperature stability. The molecular weight of the enzyme was approximately 28 kDa. Reverse micelle extraction with cetyl trimethylammonium bromide as a surfactant combined with acetone precipitation is suitable for separating and purifying fibrinolytic enzymes and a promising technique for obtaining active proteins.
研究领域:
生物化学
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酶学
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微生物学
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生物技术
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DOI
PubMed
高影响因子
Bioengineering-Basel
·
2024
·Vol. 11
·期 10
·1030
·
IF 3.8
Tuo Song,Xiaozhen Diao,Jun Cheng,Yang Man,Boyu Chen,Haixing Zhang,Wenhui Wu
As an essential thrombolytic agent, the tissue plasminogen activator receives increasing attention due to its longer half-life, lower immunogenicity, and easier administration, which are superior to other thrombolytic agents. In this study, the isolated and purified plasminogen activator from the sandworm (Perinereis aibuhitensis) was expressed inE.coli(Escherichia coli) to investigate its potential for simplifying the development process. The sandworm plasminogen activator was previously successfully cloned and expressed inE.coliwith low yield and activity in the culture supernatant. This low yield and activity prompted us to optimize its DNA sequence. Furthermore, to raise the efficiency in the separation of the target protein, the protein’s solubility was enhanced by fusing it with maltose-binding protein (MBP) tags. Eventually, the fibrinolytic activity was successfully restored after digestion with tobacco etch virus (TEV) protease. This study provides an innovative method of efficiently expressing and purifying plasminogen activators from sandworm inE.coliand broadens its applications in therapeutic treatment of cardiovascular diseases, including thrombosis, stroke, and coronary atherosclerotic heart disease.
研究领域:
生物化学
·
分子生物学
·
心血管疾病治疗
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DOI
高影响因子
NEUROPHARMACOLOGY
·
2017
·
IF 5.012
Mei-Xue Dong,Chen-Meng Li,Peng Shen,Qing-Chuan Hu,You-Dong Wei,Yi-Fei Ren,Jia Yu,Si-Wen Gui,Yi-Yun Liu,Jun-Xi Pan,Peng Xie
Objectives Recombinant tissue plasminogen activator (rtPA) is widely used for patients with thromboembolic disease , and increasing evidence indicates that it can directly induce neurotoxicity independent of its thrombolysis property. Here, we aimed to confirm the long-term effect of rtPA on animal's behavior , and investigate the underlying pathogenesis. Methods and results Male Sprague-Dawley rats randomly received a dose of rtPA (10 mg/kg) or sterile saline. Three months later, the animals receiving rtPA displayed anxiety-like behaviors in the open field and novelty-suppressed feeding tests. To investigate the possible pathogenesis, gas chromatography-mass spectrometry-based metabolomics analysis was performed, with 18 differential metabolites identified in the hippocampus 24 h after the treatments. Based upon these differential metabolites, a metabolite-protein integrated network was generated, which indicated that ERK1/2-glutamic acid decarboxylase (GAD) 1-γ aminobutyric acid (GABA) cascade may be related to long-term anxiety-like behaviors. The GABA levels in hippocampus were decreased 24 h post-treatment and three months later, confirmed by a high performance liquid chromatography method. We also examined the expression of GAD1 and GAD2 using western blotting or immunohistochemical staining . Levels of GAD1 were persistently decreased after treatment, while GAD2 levels, GAD1-immunoreactive, and GAD2-immunoreactive neurons showed no significant differences. The underlying pathogenesis also involved activation of ERK1/2, confirmed by increased phospho-ERK1/2 24 h post-treatment. Conclusions RtPA can induce long-term anxiety-like behaviors after a clinical injected dose. The underlying pathogenesis involves the ERK1/2-GAD1-GABA cascade in the hippocampus. This pharmacological side effect of rtPA may further exacerbate post-stroke anxiety disorder for stroke patients.
研究领域:
神经科学
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药理学
·
行为科学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Pengpeng Guan,Caijie Yuan,Jiali Li,Kun Yu,Ruiqi Xie,Enling Hu,Weiwei Ding,Guangqian Lan,Fei Lu
In the treatment of thrombosis, conventional nanocarriers inevitably have problems, such as adverse reactions and difficulties in synthesis. Inspired by the concept of ‘medicine food homology,’ we extracted and purified natural exosomes from mulberry leaves as carriers for the delivery of urokinase-type plasminogen activator (uPA) for targeted therapy. The obtained mulberry leaf exosomes (MLE) possessed a desirable hydrodynamic particle size (119.4 nm), a uniform size distribution (polydispersity index = 0.174), and a negative surface charge (−23.3 mv). Before loading with uPA, MLE were grafted with cyclic RGD (cRGD) to selectively bind activated platelets for thrombus targeting. The cytometry studies revealed that MLE@cRGD has a high thrombus targeting ability about 74.3 %. Animal tests demonstrated that the delivered uPA could dissolve clots almost completely in femoral vein thrombosis models. In addition, MLE could remodel venous microenvironments by effectively eliminating reactive oxygen species (ROS) and promoting the phenotypic transformation of macrophages from M1 to M2 for venous tissue repair.
研究领域:
药理学
·
纳米技术
·
治疗学
·
生物医学工程
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产品关联
DOI
PubMed
高影响因子
Food & Function
·
2017
·Vol. 8
·期 7
·2437-2443
·
IF 3.247
Guangpu Xue,Lihu Gong,Cai Yuan,Mingming Xu,Xu Wang,Longguang Jiang,Mingdong Huang
Quercetin is a member of the flavonoids and was previously demonstrated to inhibit trypsin-like serine proteases at micromolar potencies. Different molecular models were proposed to explain such inhibition. However, controversies remain on the molecular details of inhibition. Here, we report the X-ray crystal structure of quercetin in a complex with the urokinase-type plasminogen activator (uPA), an archetypical serine protease. The structure showed that quercetin binds to the specific substrate binding pocket (S1 pocket) of uPA mainly through its two neighboring phenolic hydroxyl groups. Our study thus provides unambiguous evidence to support quercetin binding to serine proteases and defines the molecular basis of the interaction. Our results further establish that natural products with two adjacent phenolic hydroxyl groups (or catechol) are likely to inhibit other trypsin-like serine proteases, a new mechanism formerly under-recognized.
研究领域:
生物化学
·
分子生物学
·
药理学
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2018
·
IF 5.123
Xing-Pei Liu,Jing-Shuai Chen,Chang-jie Mao,He-Lin Niu,Ji-Ming Song,Bao-Kang Jin
Herein, we established a novel ultrasensitive photoelectrochemical biosensor for detecting urokinase-type plasminogen activator (u-PA), based on a g-C 3 N 4 /CdS nanocomposite. The prepared nanocomposite was characterized by transmission electron microscopy, X-ray photoelectron spectroscopy, ultraviolet–visible absorption spectroscopy , and Fourier transform infrared spectroscopy , thus indicating that the nanocomposite was prepared successfully. In the typical process, the prepared nanocomposite was deposited on the surface of a bare FTO electrode. After being air-dried, the g-C 3 N 4 /CdS nanocomposite modified electrode was successively incubated with antibody against urokinase-type plasminogen activator and the blocking agent BSA to produce a photoelectrochemical biosensor for u-PA. In the presence of target u-PA antigen, the photocurrent response of the prepared biosensor electrode decreased significantly. The proposed novel photoelectrochemical biosensor exhibited good sensitivity, specificity, and reproducibility for u-PA detection, and a low detection limit of 33 fg mL −1 , ranging from 1 μg mL −1 –0.1 pg mL −1 . The proposed strategy should provide a promising method for detection of other biomarkers.
研究领域:
生物传感器
·
纳米技术
·
分析化学
·
生物医学工程
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF MATERIALS SCIENCE-MATERIALS IN MEDICINE
·
2025
·Vol. 36
·期 1
·1-18
·
IF 4.5
Fang Cuifu,Duan Qing,Tang Ying,Fu Lifeng,Chen Juzheng,Zhou Weimin,Liu Fengen
Venous thromboembolism ranks as the third most prevalent cardiovascular disease. Nanotechnology-based drug carriers show significant potential for thrombolytic therapy. However, existing systems face challenges, including environmental impact and limited accessibility. To address these, we developed a novel delivery system using human black hair-derived nanoparticles (HNPs). These HNPs were loaded with the thrombolytic agent urokinase plasminogen activator (uPA) and a minimal concentration of gold nanoparticles (AuNPs). These components were encapsulated within a polyvinyl alcohol (PVA) membrane to create the nanocapsule drug delivery system, uPA-Au@HNP@PVA. Upon intravenous injection into a rat model with venous thrombosis, the system leveraged targeted blood flow to reach thrombus sites. Under 1064 nm NIR irradiation, the PVA membrane dissolved, exposing and releasing the drugs for intelligent thrombolysis. This system demonstrated excellent thrombolytic efficacy both in vivo and in vitro, coupled with robust biocompatibility in various biological tests, suggesting a wide range of potential applications. Graphical Representation: a. Depiction of the uPA-Au@HNP@PVA synthesis process. b. Diagrammatic explanation of intelligent thrombolytic therapy for specific drug delivery in a rat venous thrombosis model: I. Intravenous administration of uPA-Au@HNP@PVA; II. Targeted delivery to thrombus sites, navigation of uPA-Au@HNP@PVA to the thrombus location through targeted blood circulation, whereupon exposure to 1064 nm NIR irradiation causes a temperature rise, resulting in PVA dissolution, and drug release; III. NIR-triggered drug release and thrombolysis. Intelligent thrombolysis facilitated by 1064 nm NIR irradiation.
研究领域:
纳米技术
·
生物医学工程
·
心血管医学
·
药学
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光响应治疗
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）