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1.
Proc Natl Acad Sci U S A ; 119(33): e2207200119, 2022 08 16.
Artigo em Inglês | MEDLINE | ID: mdl-35858375

RESUMO

The ability to produce folded and functional proteins is a necessity for structural biology and many other biological sciences. This task is particularly challenging for numerous biomedically important targets in human cells, including membrane proteins and large macromolecular assemblies, hampering mechanistic studies and drug development efforts. Here we describe a method combining CRISPR-Cas gene editing and fluorescence-activated cell sorting to rapidly tag and purify endogenous proteins in HEK cells for structural characterization. We applied this approach to study the human proteasome from HEK cells and rapidly determined cryogenic electron microscopy structures of major proteasomal complexes, including a high-resolution structure of intact human PA28αß-20S. Our structures reveal that PA28 with a subunit stoichiometry of 3α/4ß engages tightly with the 20S proteasome. Addition of a hydrophilic peptide shows that polypeptides entering through PA28 are held in the antechamber of 20S prior to degradation in the proteolytic chamber. This study provides critical insights into an important proteasome complex and demonstrates key methodologies for the tagging of proteins from endogenous sources.


Assuntos
Citometria de Fluxo , Edição de Genes , Proteínas Musculares , Complexo de Endopeptidases do Proteassoma , Sistemas CRISPR-Cas , Microscopia Crioeletrônica , Citometria de Fluxo/métodos , Edição de Genes/métodos , Células HEK293 , Humanos , Proteínas Musculares/química , Proteínas Musculares/genética , Proteínas Musculares/isolamento & purificação , Complexo de Endopeptidases do Proteassoma/química , Complexo de Endopeptidases do Proteassoma/genética , Complexo de Endopeptidases do Proteassoma/isolamento & purificação , Proteólise
2.
Lab Invest ; 104(3): 100326, 2024 03.
Artigo em Inglês | MEDLINE | ID: mdl-38237739

RESUMO

To better understand the pathogenesis of acute type A aortic dissection, high-sensitivity liquid chromatography-tandem mass spectrometry/mass spectrometry (LC-MS/MS)-based proteomics and phosphoproteomics approaches were used to identify differential proteins. Heat shock protein family B (small) member 6 (HSPB6) in aortic dissection was significantly reduced in human and mouse aortic dissection samples by real-time PCR, western blotting, and immunohistochemical staining techniques. Using an HSPB6-knockout mouse, we investigated the potential role of HSPB6 in ß-aminopropionitrile monofumarate-induced aortic dissection. We found increased mortality and increased probability of ascending aortic dissection after HSPB6 knockout compared with wild-type mice. Mechanistically, our data suggest that HSPB6 deletion promoted vascular smooth muscle cell apoptosis. More importantly, HSPB6 deletion attenuated cofilin activity, leading to excessive smooth muscle cell stiffness and eventually resulting in the development of aortic dissection and rupture. Our data suggest that excessive stiffness of vascular smooth muscle cells caused by HSPB6 deficiency is a new pathogenetic mechanism leading to aortic dissection.


Assuntos
Dissecção Aórtica , Espectrometria de Massas em Tandem , Camundongos , Humanos , Animais , Cromatografia Líquida , Dissecção Aórtica/genética , Miócitos de Músculo Liso/metabolismo , Camundongos Knockout , Modelos Animais de Doenças , Proteínas de Choque Térmico HSP20/metabolismo
3.
Mikrochim Acta ; 191(5): 282, 2024 04 23.
Artigo em Inglês | MEDLINE | ID: mdl-38652326

RESUMO

A novel dual-mode fluorometric and colorimetric sensing platform is reported for determining glutathione S-transferase (GST) by utilizing polyethyleneimine-capped silver nanoclusters (PEI-AgNCs) and cobalt-manganese oxide nanosheets (CoMn-ONSs) with oxidase-like activity. Abundant active oxygen species (O2•-) can be produced through the CoMn-ONSs interacting with dissolved oxygen. Afterward, the pink oxDPD was generated through the oxidation of colorless N,N-diethyl-p-phenylenediamine (DPD) by O2•-, and two absorption peaks at 510 and 551 nm could be observed. Simultaneously, oxDPD could quench the fluorescence of PEI-AgNCs at 504 nm via the inner filter effect (IFE). However, in the presence of glutathione (GSH), GSH prevents the oxidation of DPD due to the reducibility of GSH, leading to the absorbance decrease at 510 and 551 nm. Furthermore, the fluorescence at 504 nm was restored due to the quenching effect of oxDPD on decreased PEI-AgNCs. Under the catalysis of GST, GSH and1-chloro-2,4-dinitrobenzo (CDNB) conjugate to generate an adduct, initiating the occurrence of the oxidation of the chromogenic substrate DPD, thereby inducing a distinct colorimetric response again and the significant quenching of PEI-AgNCs. The detection limits for GST determination were 0.04 and 0.21 U/L for fluorometric and colorimetric modes, respectively. The sensing platform illustrated reliable applicability in detecting GST in real samples.


Assuntos
Cobalto , Colorimetria , Glutationa Transferase , Compostos de Manganês , Nanopartículas Metálicas , Óxidos , Polietilenoimina , Prata , Polietilenoimina/química , Prata/química , Cobalto/química , Óxidos/química , Compostos de Manganês/química , Nanopartículas Metálicas/química , Colorimetria/métodos , Glutationa Transferase/metabolismo , Glutationa Transferase/química , Limite de Detecção , Oxirredutases/química , Oxirredutases/metabolismo , Humanos , Glutationa/química , Oxirredução , Técnicas Biossensoriais/métodos , Fenilenodiaminas/química , Nanoestruturas/química
4.
J Am Chem Soc ; 145(43): 23639-23650, 2023 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-37850865

RESUMO

Photoanodes used in a water-splitting photoelectrochemical cell are almost always paired with an oxygen evolution catalyst (OEC) to efficiently utilize photon-generated holes for water oxidation because the surfaces of photoanodes are typically not catalytic for the water oxidation reaction. Suppressing electron-hole recombination at the photoanode/OEC interface is critical for the OEC to maximally utilize the holes reaching the interface for water oxidation. In order to explicitly demonstrate and investigate how the detailed features of the photoanode/OEC interface affect interfacial charge transfer and photocurrent generation for water oxidation, we prepared two BiVO4(010)/FeOOH photoanodes with different Bi:V ratios at the outermost layer of the BiVO4 interface (close to stoichiometric vs Bi-rich) while keeping all other factors in the bulk BiVO4 and FeOOH layers identical. The resulting two photoanodes show striking differences in the photocurrent onset potential and photocurrent density for water oxidation. The ambient pressure X-ray photoelectron spectroscopy results show that these two BiVO4(010)/FeOOH photoanodes show drastically different Fe2+:Fe3+ ratios in FeOOH both in the dark and under illumination with water, demonstrating the immense impact of the interfacial composition and structure on interfacial charge transfer. Using computational studies, we reveal the effect of the surface Bi:V ratio on the hydration of the BiVO4 surface and bonding with the FeOOH layer, which in turn affect the band alignments between BiVO4 and FeOOH. These results explain the atomic origin of the experimentally observed differences in electron and hole transfer and solar water oxidation performance of the two photoanodes having different interfacial compositions.

5.
Nanotechnology ; 34(12)2023 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-36538812

RESUMO

Discovery of structure-property relationships in thin film alloys of complex metal oxides enabled by high-throughput materials synthesis and characterization facilities is demonstrated here with a case-study. Thin films of binary transition metal oxides (Ti-Zn) are prepared by pulsed laser deposition with continuously varying Ti:Zn ratio, creating combinatorial samples for exploration of the properties of this material family. The atomic structure and electronic properties are probed by spatially resolved techniques including x-ray absorption near edge structures (XANES) and x-ray fluorescence (XRF) at the Ti and Zn K-edge, x-ray diffraction, and spectroscopic ellipsometry. The observed properties as a function of Ti:Zn ratio are resolved into mixtures of five distinguishable phases by deploying multivariate curve resolution analysis on the XANES spectral series, under constraints set by results from the other characterization techniques. First-principles computations based on density function theory connect the observed properties of each distinct phase with structural and spectral characteristics of crystalline polymorphs of Ti-Zn oxide. Continuous tuning of the optical absorption edge as a function of Ti:Zn ratio, including the unusual observation of negative optical bowing, exemplifies a functional property of the film correlated to the phase evolution.

6.
Mikrochim Acta ; 190(11): 444, 2023 10 18.
Artigo em Inglês | MEDLINE | ID: mdl-37851103

RESUMO

A multi-signal aptasensor for thrombin determination is proposed based on catalytically active gold nanoparticles (AuNPs) and fluorescent silicon quantum dots (SiQDs). Yellow 4-Nitrophenol (4-NP) could be converted to colorless 4-Aminophenol (4-AP) by catalytically active aptamer-modified AuNPs (S1-AuNPs). The SiQDs emitted strong blue fluorescence at 455 nm at the excitation wavelength of 367 nm. When thrombin was absent, S1-AuNPs could catalytically reduce yellow 4-NP to colorless 4-AP. When thrombin was added, the aptamer could be transformed into a G-quadruplex structure, which masked the surface-active catalytic sites of AuNPs and restrained the reduction of 4-NP. Thus, the fluorescence of SiQDs was greatly quenched by 4-NP through the inner filter effect (IFE), and the solution color remained yellow. As the concentration of thrombin increased, the catalytic activity of S1-AuNPs decreased. The concentration of 4-NP that was converted to 4-AP declined and the unconverted 4-NP increased. In this process, the absorption peak of 4-NP at 400 nm increased while the fluorescence emission of SiQDs at 455 nm decreased. The linear ranges of the fluorometric and colorimetric aptasensor were 0.5-30 nM and 0.3-30 nM, respectively. The limits of detection (LOD) for the two modes were 0.15 nM and 0.13 nM. Furthermore, a portable sensing platform was constructed by combining the smartphone-based device with the software ImageJ for the determination of thrombin. With the advantages of cost-effectiveness, simplicity of operation and broad applicability, this aptasensor provided a new perspective for on-site determination of thrombin in the clinical field.


Assuntos
Aptâmeros de Nucleotídeos , Nanopartículas Metálicas , Pontos Quânticos , Pontos Quânticos/química , Ouro/química , Trombina , Silício , Nanopartículas Metálicas/química , Aptâmeros de Nucleotídeos/química , Corantes
7.
Anal Chem ; 94(38): 13261-13268, 2022 09 27.
Artigo em Inglês | MEDLINE | ID: mdl-36103467

RESUMO

Inspired by the architecture of single-atom catalysts, where the monodispersed metal atoms are widely distributed but stabilized by various coordination circumstances, the biomimetic design and synthesis of metalloporphyrin-containing nanocages have been demonstrated in this study. The nanocages were fabricated through a coordination-driven self-assembly process, and the Mn(III) porphyrin-based one was found to have exclusively peroxidase-like activity at pH 6.0 with neither oxidase nor catalase-like activity under the routine conditions. Benefiting from this, we demonstrated the wide applicability and convenient usage of an Mn(III)-containing supramolecular nanocage (Mn-PC) in the one-step detection of H2O2, sarcosine, and glucose through various oxidase-involved reactions, with a satisfactory detection limit and eligible specificity. Real samples including H2O2 in lens care solution, sarcosine in human urine, and glucose in human serum were also assayed, showing an adequate recovery rate. Such a specific activity originates from the super-consistent microstructure of each catalytic unit, which means that the active site of manganese porphyrin was "protected" by the confinement of the nanocage. This also helps to sustain the super long-term activity even after 545 days of storage. Furthermore, the intrinsic electronic structure of the Mn(III)-containing supramolecular nanocage endows the ability in electrochemical detection of H2O2 and glucose. Our smart design toward the supramolecular nanocages with a defined structure and quantity contributes to the construction of the ingenious sensing platform and has guiding significance for architectural design of nanozymes.


Assuntos
Metaloporfirinas , Porfirinas , Catalase , Domínio Catalítico , Glucose/química , Humanos , Peróxido de Hidrogênio/química , Manganês/química , Metaloporfirinas/química , Peroxidase/metabolismo , Peroxidases , Porfirinas/química , Sarcosina
8.
Exp Dermatol ; 31(9): 1421-1430, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35598279

RESUMO

Oral lichen planus (OLP) is a T cell-mediated, chronic inflammatory disease. CD4+ T-cell infiltration plays a crucial role in the pathogenesis of OLP. Fibroblasts are activated under pathological conditions and perform various functions. This study was designed to explore the immune activation and biological functions of OLP fibroblasts on CD4+ T cells. We detected the expression of fibroblast activation protein-alpha (FAP-α) in the oral tissues of patients with OLP and healthy controls using immunohistochemistry. Furthermore, expression of FAP-α and C-C motif chemokine ligand 5 (CCL5) in fibroblasts isolated from oral tissues of patients with OLP and healthy controls was assayed by quantitative PCR, Western blotting and enzyme-linked immunosorbent assay. Moreover, we assessed the effects of fibroblasts on CD4+ T-cell proliferation, apoptosis and migration using flow cytometry and Transwell assays. We found that FAP-α expression in the oral tissues of patients with OLP was significantly higher than that in healthy controls. FAP-α and CCL5 expression levels were significantly upregulated in OLP fibroblasts. Moreover, OLP fibroblasts promoted CD4+ T-cell proliferation and migration and inhibited CD4+ T cell apoptosis. In summary, our findings indicate that OLP fibroblasts are immunologically activated and induce CD4+ T-cell infiltration in OLP.


Assuntos
Líquen Plano Bucal , Linfócitos T CD4-Positivos/metabolismo , Quimiocina CCL5/metabolismo , Endopeptidases , Humanos , Líquen Plano Bucal/metabolismo , Proteínas de Membrana , Linfócitos T
9.
Mikrochim Acta ; 189(4): 135, 2022 03 07.
Artigo em Inglês | MEDLINE | ID: mdl-35257215

RESUMO

Iron-cobalt oxide nanosheets (FeCo-ONSs) were proved to have intrinsic peroxidase-like activity. Additionally, the peroxidase-like activity of FeCo-ONSs toward the oxidation of 3,3,5,5-tetramethylbenzidine (TMB) was dramatically enhanced after heparin addition due to the stronger affinity toward TMB. Protamine combines with heparin, so the promotion of peroxidase-like activity of FeCo-ONSs with heparin was suppressed. With the addition of trypsin, protamine was hydrolyzed and the enhancement effect of catalytic activity of FeCo-ONSs was recovered. Based on above process, a sensitive colorimetric platform for trypsin activity determination was constructed through measuring the absorbance of produced oxTMB at 652 nm, providing a linear detection range of 5 to 500 ng/mL and a low detection limit of 2.8 ng/mL. The method was applied to trypsin determination in real samples (human urine sample and multienzyme tablet sample) with satisfactory results, illustrating the potential application of this biosensor.


Assuntos
Colorimetria , Peroxidase , Cobalto , Colorimetria/métodos , Heparina , Humanos , Ferro , Óxidos , Oxirredutases , Peroxidases , Protaminas , Tripsina
10.
Molecules ; 27(12)2022 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-35744798

RESUMO

Phillyrin is an effective lignan glycoside extracted from a traditional Chinese medicine Forsythia suspensa (Thunb.) Vahl (Oleaceae). It mainly exists in the roots, stems, leaves and fruits of the plant, with the highest content in the leaves. In terms of its medicinal application, there are a large number of experimental data proving its pharmacological effects in vitro and in animal models, such as anti-inflammatory, anti-obesity, anti-tumor, etc. Furthermore, pharmacokinetic experiments have also shown phillyrin's high effectiveness and low toxicity. Despite more than one thousand studies in the literature on phillyrin retrievable from Web of Science, PubMed, and CNKI, few reviews on its pharmacological activities have been presented conclusively. In this paper, we aimed to summarize the pharmacological and pharmacokinetic characteristics of phillyrin from the current literature, focusing on its anti-inflammatory, anti-aging, antiviral, antibacterial, hepatoprotective and anti-cancer effects, hoping to come up with new insights for its application as well as future studies.


Assuntos
Forsythia , Animais , Anti-Inflamatórios/farmacologia , Glucosídeos/farmacologia , Glicosídeos , Extratos Vegetais/farmacologia
12.
Talanta ; 277: 126400, 2024 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-38876031

RESUMO

Captopril (CP) is commonly used as an active enzyme inhibitor for the treatment of coronary heart disease, hypertension and angina pectoris. The development of sensitive and efficient method for CP analysis is of great importance in biomedical research. Herein, we fabricated a sensitive and robust hydrogel-assisted paper-based sensor based on fluorescence UiO-66-NH2@ZIF-8 and Co, N-doped carbon nanozymes with oxidase-mimicking activity for accurate monitoring of captopril. The hydrogel-assisted paper-based sensor appeared a visible pink signal due to the catalytic oxidation of colorless N,N-diethyl-p-phenylenediamine (DPD) to oxDPD by Co, N-doped carbon-based nanozymes, and resulted in the fluorescence quenching of UiO-66-NH2@ZIF-8. In the presence of captopril, the oxidation of chromogenic substrate DPD by Co, N-doped nanozymes in the hydrogel-assisted paper-based sensor was hindered and accompanied by a change in the visible color, leading to recovery of the fluorescence of UiO-66-NH2@ZIF-8, and the change in the fluorescence color could also be observed. Therefore, the quantitative detection of captopril is achieved by taking a smartphone photograph and converting the image parameters into data information using ImageJ software. The portable hydrogel-assisted paper sensor provided sensitive detection of captopril in two modes based on visible color change as well as fluorescence color change with limits of detection of 0.45 µM and 0.47 µM, respectively. This hydrogel-assisted paper-based sensor has been successfully applied to the accurate monitoring of captopril in human serum, providing a potential avenue for in situ detection of captopril.

13.
Anal Chim Acta ; 1311: 342715, 2024 Jul 04.
Artigo em Inglês | MEDLINE | ID: mdl-38816154

RESUMO

BACKGROUND: Due to that the higher activity of nanozymes would bring outstanding performance for the nanozyme-based biosensing strategies, great efforts have been made by researchers to improve the catalytic activity of nanozymes, and novel nanozymes with high catalytic activity are desired. Considering the crucial role in controlling blood glucose level, strategies like colorimetric and chemiluminescence to monitor α-glucosidase are developed. However, multi-mode detection with higher sensitivity was insufficient. Therefore, developing triple-mode detection method for α-glucosidase based on great performance nanozyme is of great importance. RESULTS: In this work, a novel nanozyme Cu-BCN was synthesized by loading Cu on boron doped carbon substrate g-C3N4 and applied to the colorimetric-fluorescent-smartphone triple-mode detection of α-glucosidase. In the presence of H2O2, Cu-BCN catalyzed the generation of 1O2 from H2O2, 1O2 subsequently oxidized TMB to blue colored oxTMB. In the presence of hydroquinone (HQ), the ROS produced from H2O2 was consumed, inhibiting the oxidation of TMB, which endows the possibility of colorimetric and visual on-site detection of HQ. Further, due to that the fluorescence of Mg-CQDs at 444 nm could be quenched by oxTMB, HQ could also be quantified through fluorescent mode. Since α-glucosidase could efficiently hydrolyze α-arbutin into HQ, the sensitive detection of α-glucosidase was realized. Further, colorimetric paper-based device (c-PAD) was fabricated for on-site α-glucosidase detection. The LODs for α-glucosidase via three modes were 2.20, 1.62 and 2.83 U/L respectively, high sensitivities were realized. SIGNIFICANCE: The nanozyme Cu-BCN possesses higher peroxidase-like activity by doping boron to the substrate than non-doped Cu-CN. The proposed triple-mode detection of α-glucosidase is more sensitive than most previous reports, and is reliable when applied to practical sample. Further, the smartphone-based colorimetric paper-based analytical device (c-PAD) made of simple materials could also detect α-glucosidase sensitively. The smartphone-based on-site detection provided a convenient, instrument-free and sensitive sensing method for α-glucosidase.


Assuntos
Boro , Colorimetria , Cobre , Smartphone , alfa-Glucosidases , Colorimetria/métodos , Cobre/química , alfa-Glucosidases/metabolismo , alfa-Glucosidases/química , Boro/química , Compostos de Nitrogênio/química , Limite de Detecção , Técnicas Biossensoriais/métodos , Corantes Fluorescentes/química , Humanos , Grafite
14.
Food Chem ; 441: 138372, 2024 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-38219364

RESUMO

Herein, we synthesized a novel N-doped carbon layer encapsulated Fe/Co bimetallic nanoparticles (Fe/Co-NC), which exhibited superior oxidase-like activity due to the facilitation of electron penetration and the formation of metal-nitrogen active sites. Fe/Co-NC could catalyze the oxidation of 3,3,5,5-tetramethylbenzidine (TMB) to blue oxTMB. Acetylcholinesterase (AChE) could catalyze the hydrolysis of thioacetylcholine to produce reducing thiocholine, which prevented TMB from oxidation. Thus, a portable hydrogel colorimetric sensor was developed for on-site and visual monitoring of AChE with the detection limit of 0.36 U L-1, and successfully applied to detect AChE in human erythrocyte samples. Furthermore, this platform was used to investigate the inhibition of triazophos on AChE activity.


Assuntos
Benzidinas , Praguicidas , Humanos , Praguicidas/análise , Oxirredutases/química , Acetilcolinesterase , Colorimetria , Hidrogéis
15.
Int J Biol Macromol ; 260(Pt 2): 129580, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38246442

RESUMO

Adding bio-based flame retardants to improve the flame retardancy of polymer materials without sacrificing other properties is a great challenge. Herein, a novel flame-retardant CS-DOPA was prepared from chitosan and 10-hydroxy-9,10-dihydro-9-oza-10-phosphaphenanthrene-10-oxide by acid-base neutralization reaction and fully characterized. The 4 wt% CS-DOPA modified EP showed good flame retardancy in both gaseous and condensed phase. The peak heat release rate, total smoke production, CO production, and smoke production rate of EP composites containing 4 wt% CS-DOPA were reduced by 55 %, 34 %, 45 %, and 46 %, respectively, to pass the UL-94 V-1 rating with a limiting oxygen index of 34.1 %. The CS-DOPA contributes to the formation of the condensed phase of the thermo-oxidation-resistant high-quality char layer with non-flammable other and phosphorus-containing free radicals released in the gas phase. In addition, EP/4CS-DOPA has good water resistance, mechanical properties, and transparency, with tensile and flexural strength improved by 12.7 % and 13.9 %, respectively, and still has high strength even after water treatment. The present work provides a green and facile strategy to use chitosan as a main raw material to manufacture EP materials with high performance.


Assuntos
Quitosana , Retardadores de Chama , Resinas Epóxi , Gases , Di-Hidroxifenilalanina
16.
ACS Nano ; 18(1): 1126-1136, 2024 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-38147003

RESUMO

Recent advances in superconducting qubit technology have led to significant progress in quantum computing, but the challenge of achieving a long coherence time remains. Despite the excellent lifetime performance that tantalum (Ta) based qubits have demonstrated to date, the majority of superconducting qubit systems, including Ta-based qubits, are generally believed to have uncontrolled surface oxidation as the primary source of the two-level system loss in two-dimensional transmon qubits. Therefore, atomic-scale insight into the surface oxidation process is needed to make progress toward a practical quantum processor. In this study, the surface oxidation mechanism of native Ta films and its potential impact on the lifetime of superconducting qubits were investigated using advanced scanning transmission electron microscopy (STEM) techniques combined with density functional theory calculations. The results suggest an atomistic model of the oxidized Ta(110) surface, showing that oxygen atoms tend to penetrate the Ta surface and accumulate between the two outermost Ta atomic planes; oxygen accumulation at the level exceeding a 1:1 O/Ta ratio drives disordering and, eventually, the formation of an amorphous Ta2O5 phase. In addition, we discuss how the formation of a noninsulating ordered TaO1-δ (δ < 0.1) suboxide layer could further contribute to the losses of superconducting qubits. Subsurface oxidation leads to charge redistribution and electric polarization, potentially causing quasiparticle loss and decreased current-carrying capacity, thus affecting superconducting qubit coherence. The findings enhance the comprehension of the realistic factors that might influence the performance of superconducting qubits, thus providing valuable guidance for the development of future quantum computing hardware.

17.
Front Pharmacol ; 15: 1288584, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38500762

RESUMO

Objective: To evaluate the efficacy of the fruits of the medicinal plant Forsythia suspensa (Thunb.) Vahl (FS), in treating inflammation-associated diseases through a meta-analysis of animal models, and also probe deeply into the signaling pathways underlying the progression of inflammation. Materials and methods: All data analyses were performed using Review Manager 5.3 and the results are presented as flow diagrams, risk-of-bias summaries, forest plots, and funnel plots. Summary estimates were calculated using a random- or fixed-effect model, depending on the value of I2. Results: Of the 710 records identified in the initial search, 11 were selected for the final meta-analysis. Each study extracted data from the model and treatment groups for analysis, and the results showed that FS alleviated the inflammatory cytokine levels in serum; oxidant indicator: reactive oxygen species; enzymes of liver function; endotoxin and regulatory cells in blood; and improved the antioxidant enzyme superoxide dismutase. Conclusion: FS effectively reversed the change in acute or chronic inflammation indicators in animal models, and the regulation of multiple channel proteins in inflammatory signaling pathways suggests that FS is a good potential drug for inflammatory disease drug therapy.

18.
J Mol Model ; 30(3): 68, 2024 Feb 13.
Artigo em Inglês | MEDLINE | ID: mdl-38347278

RESUMO

CONTEXT: Adipose triglyceride lipase (ATGL), a key enzyme responsible for lipolysis, catalyzes the first step of lipolysis and converts triglycerides to diacylglycerols and free fatty acids (FFA). Our previous work suggested that phillyrin treatment improves insulin resistance in HFD-fed mice, which was associated with ATGL inhibition. In this study, using docking simulation, we explored the binding pose of phillyrin and atglistatin (a mouse ATGL inhibitor) to ATGL in mouse. From the docking results, the interactions with Ser47 and Asp166 were speculated to have caused phillyrin to inhibit ATGL in mice. Further, molecular dynamics simulation of 100 ns and MM-GBSA were conducted for the protein-ligand complex, which indicated that the system was stable and that phillyrin displayed a better affinity to ATGL than did atglistatin throughout the simulation period. Moreover, the results of pharmacological validation were consistent with those of the in silico simulations. In summary, our study illustrates the potential of molecular docking to accurately predict the binding protein produced by AlphaFold and suggests that phillyrin is a potential small molecule that targets and inhibits ATGL enzymatic activity. METHODS: The ATGL-predicted protein structure, verified by PROCHECK, was determined using AlphaFold. Molecular docking, molecular dynamics simulation, and prime molecular mechanic-generalized born surface area were performed using LigPrep, Desmond, and prime MM-GBSA modules of Schrödinger software release 2021-2, respectively. For pharmacological validation, immunoblotting was performed to assess ATGL protein expression. The fluorescence intensity and glycerol concentration were quantified to evaluate the efficiency of phillyrin in inhibiting ATGL.


Assuntos
Glucosídeos , Lipase , Lipólise , Compostos de Fenilureia , Camundongos , Animais , Simulação de Acoplamento Molecular , Lipase/metabolismo , Lipólise/fisiologia
19.
Heliyon ; 10(9): e30323, 2024 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-38711632

RESUMO

Background: Prolonged circulatory arrest time is an independent risk factor for postoperative adverse events of type A aortic dissection (TAAD) surgery. Further reduction of the circulatory arrest time is essential to improve surgical outcomes. This study aimed to evaluate the safety and effectiveness of the novel Sutureless Integrated Stented (SIS) graft prosthesis in an animal experiment. Materials and methods: Straight type of the SIS graft prosthesis was implanted into the descending aorta of 10 adult male sheep, and the use of the device was scored on a scale of 1-10. Aortic digital subtraction angiography (DSA) was performed at 4, 14, and 26 weeks to investigate the prostheses. After 26 weeks, the animals were sacrificed for histological analysis. Results: The immediate success rate of the surgery was 100 %, and the overall mean score of the use of the device was 9.65 ± 0.99. Three animals died from non-device-related causes during follow-up. Aortic DSA showed filling defects in 5 animals. Histological analysis revealed that all prostheses were intact. Except for 2 early deaths, the other 8 prostheses were endothelialized with mild inflammation, foreign body reactions, and intimal fibrosis. The mean cross-sectional area of the sutureless region was reduced by 26.4 % (range, 1.3-39.1 %). Conclusions: The safety and effectiveness of the novel SIS graft prosthesis were acceptable, and the delivery system exhibited a promising performance. Using the SIS graft prosthesis in TAAD surgery was expected to simplify the procedures and shorten the circulatory arrest time. Further large-scale clinical trials are required to verify these findings.

20.
Metabolites ; 14(4)2024 Apr 21.
Artigo em Inglês | MEDLINE | ID: mdl-38668372

RESUMO

Rice (Oryza sativa L.), a crucial food crop that sustains over half the world's population, is often hindered by salt stress during various growth stages, ultimately causing a decrease in yield. However, the specific mechanism of rice roots' response to salt stress remains largely unknown. In this study, transcriptomics and lipidomics were used to analyze the changes in the lipid metabolism and gene expression profiles of rice roots in response to salt stress. The results showed that salt stress significantly inhibited rice roots' growth and increased the roots' MDA content. Furthermore, 1286 differentially expressed genes including 526 upregulated and 760 downregulated, were identified as responding to salt stress in rice roots. The lipidomic analysis revealed that the composition and unsaturation of membrane lipids were significantly altered. In total, 249 lipid molecules were differentially accumulated in rice roots as a response to salt stress. And most of the major phospholipids, such as phosphatidic acid (PA), phosphatidylcholine (PC), and phosphatidylserine (PS), as well as major sphingolipids including ceramide (Cer), phytoceramide (CerP), monohexose ceramide (Hex1Cer), and sphingosine (SPH), were significantly increased, while the triglyceride (TG) molecules decreased. These results suggested that rice roots mitigate salt stress by altering the fluidity and integrity of cell membranes. This study enhances our comprehension of salt stress, offering valuable insights into changes in the lipids and adaptive lipid remodeling in rice's response to salt stress.

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