Your browser doesn't support javascript.
loading
Montrer: 20 | 50 | 100
Résultats 1 - 20 de 1.206
Filtrer
1.
Biopharm Drug Dispos ; 45(3): 149-158, 2024 Jun.
Article de Anglais | MEDLINE | ID: mdl-38886878

RÉSUMÉ

Icaritin is a prenylflavonoid derivative of the genus Epimedium (Berberidaceae) and has a variety of pharmacological actions. Icaritin is approved by the National Medical Products Administration as an anticancer drug that exhibits efficacy and safety advantages in patients with hepatocellular carcinoma cells. This study aimed to evaluate the inhibitory effects of icaritin on UDP-glucuronosyltransferase (UGT) isoforms. 4-Methylumbelliferone (4-MU) was employed as a probe drug for all the tested UGT isoforms using in vitro human liver microsomes (HLM). The inhibition potentials of UGT1A1 and 1A9 in HLM were further tested by employing 17ß-estradiol (E2) and propofol (PRO) as probe substrates, respectively. The results showed that icaritin inhibits UGT1A1, 1A3, 1A4, 1A7, 1A8, 1A10, 2B7, and 2B15. Furthermore, icaritin exhibited a mixed inhibition of UGT1A1, 1A3, and 1A9, and the inhibition kinetic parameters (Ki) were calculated to be 3.538, 2.117, and 0.306 (µM), respectively. The inhibition of human liver microsomal UGT1A1 and 1A9 both followed mixed mechanism, with Ki values of 2.694 and 1.431 (µM). This study provides supporting information for understanding the drug-drug interaction (DDI) potential of the flavonoid icaritin and other UGT-metabolized drugs in clinical settings. In addition, the findings provide safety evidence for DDI when liver cancer patients receive a combination therapy including icaritin.


Sujet(s)
Interactions médicamenteuses , Flavonoïdes , Glucuronosyltransferase , Microsomes du foie , Glucuronosyltransferase/antagonistes et inhibiteurs , Glucuronosyltransferase/métabolisme , Humains , Flavonoïdes/pharmacologie , Microsomes du foie/métabolisme , Oestradiol/pharmacologie , Hymécromone/pharmacologie , Propofol/pharmacologie , Antienzymes/pharmacologie
2.
ACS Appl Mater Interfaces ; 16(22): 28222-28229, 2024 Jun 05.
Article de Anglais | MEDLINE | ID: mdl-38779815

RÉSUMÉ

ß-Glucosidase (EC 3.2.1.21) from sweet almond was encapsulated into pH-responsive alginate-polyethylenimine (alginate-PEI) hydrogel. Then, electrochemically controlled cyclic local pH changes resulting from ascorbate oxidation (acidification) and oxygen reduction (basification) were used for the pulsatile release of the enzyme from the composite hydrogel. Activation of the enzyme was controlled by the very same pH changes used for ß-glucosidase release, separating these two processes in time. Importantly, the activity of the enzyme, which had not been released yet, was inhibited due to the buffering effect of PEI present in the gel. Thus, only a portion of the released enzyme was activated. Both enzymatic activity and release were monitored by confocal fluorescence microscopy and regular fluorescent spectroscopy. Namely, commercially available very little or nonfluorescent substrate 4-methylumbelliferyl-ß-d-glucopyranoside was hydrolyzed by ß-glucosidase to produce a highly fluorescent product 4-methylumbelliferone during the activation phase. At the same time, labeling of the enzyme with rhodamine B isothiocyanate was used for release observation. The proposed work represents an interesting smart release-activation system with potential applications in biomedical field.


Sujet(s)
Alginates , Hydrogels , Polyéthylèneimine , bêta-Glucosidase , Alginates/composition chimique , Hydrogels/composition chimique , Polyéthylèneimine/composition chimique , Concentration en ions d'hydrogène , bêta-Glucosidase/métabolisme , bêta-Glucosidase/composition chimique , Rhodamines/composition chimique , Enzymes immobilisées/composition chimique , Enzymes immobilisées/métabolisme , Hymécromone/composition chimique , Activation enzymatique/effets des médicaments et des substances chimiques , Prunus/enzymologie , Prunus/composition chimique , Acide glucuronique/composition chimique , Techniques électrochimiques
3.
Int J Biol Macromol ; 268(Pt 2): 131697, 2024 May.
Article de Anglais | MEDLINE | ID: mdl-38688333

RÉSUMÉ

Immobilization technology plays an important role in enhancing enzyme stability and environmental adaptability. Despite its rapid development, this technology still encounters many challenges such as enzyme leakage, difficulties in large-scale implementation, and limited reusability. Drawing inspiration from natural paired molecules, this study aimed to establish a method for immobilized α-glucosidase using artificial antibody-antigen interaction. The proposed method consists of three main parts: synthesis of artificial antibodies, synthesis of artificial antigens, and assembly of the artificial antibody-antigen complex. The critical step in this method involves selecting a pair of structurally similar compounds: catechol as a template for preparing artificial antibodies and protocatechualdehyde for modifying the enzyme to create the artificial antigens. By utilizing the same functional groups in these compounds, specific recognition of the antigen by the artificial antibody can be achieved, thereby immobilizing the enzymes. The results demonstrated that the immobilization amount, specific activity, and enzyme activity of the immobilized α-glucosidase were 25.09 ± 0.10 mg/g, 5.71 ± 0.17 U/mgprotein and 143.25 ± 1.71 U/gcarrier, respectively. The immobilized α-glucosidase not only exhibited excellent reusability but also demonstrated remarkable performance in catalyzing the hydrolysis of 4-methylumbelliferyl-α-D-glucopyranoside.


Sujet(s)
Enzymes immobilisées , Hymécromone , alpha-Glucosidase , Enzymes immobilisées/composition chimique , alpha-Glucosidase/composition chimique , alpha-Glucosidase/immunologie , Hymécromone/composition chimique , Hymécromone/analogues et dérivés , Biocatalyse , Stabilité enzymatique , Hydrolyse , Biomimétique/méthodes , Cinétique , Anticorps/composition chimique , Anticorps/immunologie , Matériaux biomimétiques/composition chimique , Complexe antigène-anticorps/composition chimique , Concentration en ions d'hydrogène
4.
Sci Rep ; 14(1): 2797, 2024 02 02.
Article de Anglais | MEDLINE | ID: mdl-38307876

RÉSUMÉ

Hepatic fibrosis remains a significant clinical challenge due to ineffective treatments. 4-methylumbelliferone (4MU), a hyaluronic acid (HA) synthesis inhibitor, has proven safe in phase one clinical trials. In this study, we aimed to ameliorate liver fibrosis by inhibiting HA synthesis. We compared two groups of mice with CCl4-induced fibrosis, treated with 4-methylumbelliferone (4MU) and hyaluronan synthase 2 (HAS2) targeting siRNA (siHAS2). The administration of 4MU and siHAS2 significantly reduced collagen and HA deposition, as well as biochemical markers of hepatic damage induced by repeated CCl4 injections. The transcriptomic analysis revealed converging pathways associated with downstream HA signalling. 4MU- and siHAS2-treated fibrotic livers shared 405 upregulated and 628 downregulated genes. These genes were associated with xenobiotic and cholesterol metabolism, mitosis, endoplasmic reticulum stress, RNA processing, and myeloid cell migration. The functional annotation of differentially expressed genes (DEGs) in siHAS2-treated mice revealed attenuation of extracellular matrix-associated pathways. In comparison, in the 4MU-treated group, DEGs were related to lipid and bile metabolism pathways and cell cycle. These findings confirm that HAS2 is an important pharmacological target for suppressing hepatic fibrosis using siRNA.


Sujet(s)
Acide hyaluronique , Hymécromone , Animaux , Souris , Analyse de profil d'expression de gènes , Hyaluronan synthases/génétique , Hyaluronan synthases/métabolisme , Acide hyaluronique/métabolisme , Hymécromone/pharmacologie , Cirrhose du foie/induit chimiquement , Cirrhose du foie/traitement médicamenteux , Cirrhose du foie/génétique , Petit ARN interférent
5.
Molecules ; 29(2)2024 Jan 10.
Article de Anglais | MEDLINE | ID: mdl-38257254

RÉSUMÉ

A representative naturally occurring coumarin, 4-methylumbelliferone (5), was exposed to 50 kGy of gamma ray, resulting in four newly generated dihydrocoumarin products 1-4 induced by the gamma irradiation. The structures of these new products were elucidated by interpretation of spectroscopic data (NMR, MS, [α]D, and UV). The unusual bisdihydrocoumarin 4 exhibited improved tyrosinase inhibitory capacity toward mushroom tyrosinase with IC50 values of 19.8 ± 0.5 µM as compared to the original 4-methylumbelliferone (5). A kinetic analysis also exhibited that the potent metabolite 4 had non-competitive modes of action. Linkage of the hydroxymethyl group in the C-3 and C-4 positions on the lactone ring probably enhances the tyrosinase inhibitory effect of 4-methylumbelliferone (5). Thus, the novel coumarin analog 4 is an interesting new class of tyrosinase inhibitory candidates that requires further examination.


Sujet(s)
Agaricales , Monophenol monooxygenase , Hymécromone , Cinétique , Coumarines/pharmacologie
6.
Sci Rep ; 13(1): 19183, 2023 11 06.
Article de Anglais | MEDLINE | ID: mdl-37932336

RÉSUMÉ

Spinal cord injury (SCI) induces the upregulation of chondroitin sulfate proteoglycans (CSPGs) at the glial scar and inhibits neuroregeneration. Under normal physiological condition, CSPGs interact with hyaluronan (HA) and other extracellular matrix on the neuronal surface forming a macromolecular structure called perineuronal nets (PNNs) which regulate neuroplasticity. 4-methylumbelliferone (4-MU) is a known inhibitor for HA synthesis but has not been tested in SCI. We first tested the effect of 4-MU in HA reduction in uninjured rats. After 8 weeks of 4-MU administration at a dose of 1.2 g/kg/day, we have not only observed a reduction of HA in the uninjured spinal cords but also a down-regulation of CS glycosaminoglycans (CS-GAGs). In order to assess the effect of 4-MU in chronic SCI, six weeks after Th8 spinal contusion injury, rats were fed with 4-MU or placebo for 8 weeks in combination with daily treadmill rehabilitation for 16 weeks to promote neuroplasticity. 4-MU treatment reduced the HA synthesis by astrocytes around the lesion site and increased sprouting of 5-hydroxytryptamine fibres into ventral horns. However, the current dose was not sufficient to suppress CS-GAG up-regulation induced by SCI. Further adjustment on the dosage will be required to benefit functional recovery after SCI.


Sujet(s)
Gliose , Traumatismes de la moelle épinière , Animaux , Rats , Protéoglycanes à chondroïtine sulfate , Gliose/anatomopathologie , Acide hyaluronique , Hymécromone/usage thérapeutique , Moelle spinale/anatomopathologie
7.
Matrix Biol ; 123: 34-47, 2023 Nov.
Article de Anglais | MEDLINE | ID: mdl-37783236

RÉSUMÉ

Pancreatic ß-cell dysfunction and death are central to the pathogenesis of type 2 diabetes (T2D). We identified a novel role for the inflammatory extracellular matrix polymer hyaluronan (HA) in this pathophysiology. Low concentrations of HA were present in healthy pancreatic islets. However, HA substantially accumulated in cadaveric islets of T2D patients and islets of the db/db mouse model of T2D in response to hyperglycemia. Treatment with 4-methylumbelliferone (4-MU), an inhibitor of HA synthesis, or the deletion of the main HA receptor CD44, preserved glycemic control and insulin concentrations in db/db mice despite ongoing weight gain, indicating a critical role for this pathway in T2D pathogenesis. 4-MU treatment and the deletion of CD44 likewise preserved glycemic control in other settings of ß-cell injury including streptozotocin treatment and islet transplantation. Mechanistically, we found that 4-MU increased the expression of the apoptosis inhibitor survivin, a downstream transcriptional target of CD44 dependent on HA/CD44 signaling, on ß-cells such that caspase 3 activation did not result in ß-cell apoptosis. These data indicated a role for HA accumulation in diabetes pathogenesis and suggested that it may be a viable target to ameliorate ß-cell loss in T2D. These data are particularly exciting, because 4-MU is already an approved drug (also known as hymecromone), which could accelerate translation of these findings to clinical studies.


Sujet(s)
Diabète de type 2 , Ilots pancréatiques , Souris , Animaux , Humains , Acide hyaluronique/métabolisme , Diabète de type 2/génétique , Hymécromone/pharmacologie , Ilots pancréatiques/métabolisme , Obésité/génétique , Antigènes CD44/génétique , Antigènes CD44/métabolisme
8.
Sci Rep ; 13(1): 9356, 2023 06 08.
Article de Anglais | MEDLINE | ID: mdl-37291120

RÉSUMÉ

Glioblastoma (GBM) is the most frequent malignant primary tumor of the CNS in adults, with a median survival of 14.6 months after diagnosis. The effectiveness of GBM therapies remains poor, highlighting the need for new therapeutic alternatives. In this work, we evaluated the effect of 4-methylumbelliferone (4MU), a coumarin derivative without adverse effects reported, in combination with temozolomide (TMZ) or vincristine (VCR) on U251, LN229, U251-TMZ resistant (U251-R) and LN229-TMZ resistant (LN229-R) human GBM cells. We determined cell proliferation by BrdU incorporation, migration through wound healing assay, metabolic and MMP activity by XTT and zymography assays, respectively, and cell death by PI staining and flow cytometry. 4MU sensitizes GBM cell lines to the effect of TMZ and VCR and inhibits metabolic activity and cell proliferation on U251-R cells. Interestingly, the lowest doses of TMZ enhance U251-R and LN229-R cell proliferation, while 4MU reverts this and even sensitizes both cell lines to TMZ and VCR effects. We showed a marked antitumor effect of 4MU on GBM cells alone and in combination with chemotherapy and proved, for the first time, the effect of 4MU on TMZ-resistant models, demonstrating that 4MU would be a potential therapeutic alternative for improving GBM therapy even on TMZ-refractory patients.


Sujet(s)
Tumeurs du cerveau , Glioblastome , Humains , Témozolomide/usage thérapeutique , Glioblastome/anatomopathologie , Hymécromone/pharmacologie , Résistance aux médicaments antinéoplasiques , Lignée cellulaire tumorale , Prolifération cellulaire , Tumeurs du cerveau/métabolisme , Antinéoplasiques alcoylants/pharmacologie , Antinéoplasiques alcoylants/usage thérapeutique , Apoptose , Tests d'activité antitumorale sur modèle de xénogreffe
9.
Biomacromolecules ; 24(5): 2278-2290, 2023 05 08.
Article de Anglais | MEDLINE | ID: mdl-37071718

RÉSUMÉ

Advanced multifunctional biomaterials are increasingly relying on clinically dictated patterns of selectivity against various biological targets. Integration of these frequently conflicting features into a single material surface may be best achieved by combining various complementary methodologies. Herein, a drug with a broad spectrum of activity, i.e., 4-methylumbelliferone (4-MU), is synthetically multimerized into water-soluble anionic macromolecules with the polyphosphazene backbone. The polymer structure, composition, and solution behavior are studied by 1H and 31P NMR spectroscopy, size-exclusion chromatography, dynamic light scattering, and UV and fluorescence spectrophotometry. To take advantage of the clinically proven hemocompatibility of fluorophosphazene surfaces, the drug-bearing macromolecule was then nanoassembled onto the surface of selected substrates in an aqueous solution with fluorinated polyphosphazene of the opposite charge using the layer-by-layer (LbL) technique. Nanostructured 4-MU-functionalized fluoro-coatings exhibited a strong antiproliferative effect on vascular smooth muscle cells (VSMCs) and fibroblasts with no cytotoxicity against endothelial cells. This selectivity pattern potentially provides the opportunity for highly desirable fast tissue healing while preventing the overgrowth of VSMCs and fibrosis. Taken together with the established in vitro hemocompatibility and anticoagulant activity, 4-MU-functionalized fluoro-coatings demonstrate potential for applications as restenosis-resistant coronary stents and artificial joints.


Sujet(s)
Cellules endothéliales , Hymécromone , Hymécromone/pharmacologie , Propriétés de surface , Polymères/pharmacologie , Matériaux revêtus, biocompatibles/composition chimique
10.
Int J Mol Sci ; 24(4)2023 Feb 14.
Article de Anglais | MEDLINE | ID: mdl-36835210

RÉSUMÉ

4-methylumbelliferone (4MU) has been suggested as a potential therapeutic agent for a wide range of neurological diseases. The current study aimed to evaluate the physiological changes and potential side effects after 10 weeks of 4MU treatment at a dose of 1.2 g/kg/day in healthy rats, and after 2 months of a wash-out period. Our findings revealed downregulation of hyaluronan (HA) and chondroitin sulphate proteoglycans throughout the body, significantly increased bile acids in blood samples in weeks 4 and 7 of the 4MU treatment, as well as increased blood sugars and proteins a few weeks after 4MU administration, and significantly increased interleukins IL10, IL12p70 and IFN gamma after 10 weeks of 4MU treatment. These effects, however, were reversed and no significant difference was observed between control treated and 4MU-treated animals after a 9-week wash-out period.


Sujet(s)
Acide hyaluronique , Hymécromone , Animaux , Rats , Acide hyaluronique/métabolisme , Hymécromone/effets indésirables , Hymécromone/usage thérapeutique , Interleukine-12
11.
Int J Mol Sci ; 24(3)2023 Jan 21.
Article de Anglais | MEDLINE | ID: mdl-36768453

RÉSUMÉ

4-methylumbelliferone (4MU) is a well-known hyaluronic acid synthesis inhibitor and an approved drug for the treatment of cholestasis. In animal models, 4MU decreases inflammation, reduces fibrosis, and lowers body weight, serum cholesterol, and insulin resistance. It also inhibits tumor progression and metastasis. The broad spectrum of effects suggests multiple and yet unknown targets of 4MU. Aiming at 4MU target deconvolution, we have analyzed publicly available data bases, including: 1. Small molecule library Bio Assay screening (PubChemBioAssay); 2. GO pathway databases screening; 3. Protein Atlas Database. We also performed comparative liver transcriptome analysis of mice on normal diet and mice fed with 4MU for two weeks. Potential targets of 4MU public data base analysis fall into two big groups, enzymes and transcription factors (TFs), including 13 members of the nuclear receptor superfamily regulating lipid and carbohydrate metabolism. Transcriptome analysis revealed changes in the expression of genes involved in bile acid metabolism, gluconeogenesis, and immune response. It was found that 4MU feeding decreased the accumulation of the glycogen granules in the liver. Thus, 4MU has multiple targets and can regulate cell metabolism by modulating signaling via nuclear receptors.


Sujet(s)
Hymécromone , Transcriptome , Souris , Animaux , Hymécromone/pharmacologie , Foie/métabolisme , Inflammation/métabolisme , Transduction du signal , Métabolisme lipidique
12.
Anal Chim Acta ; 1241: 340778, 2023 Feb 08.
Article de Anglais | MEDLINE | ID: mdl-36657871

RÉSUMÉ

Endogenous peroxynitrite plays a very important role in the regulation of life activities. However, validated tools for ONOO- tests are currently insufficient. We designed a fluorescent probe TPA-F-NO2 with a low fluorescence background in water based on the D-π-A structure for the imaging of endogenous ONOO- in living cells. TPA-F-NO2 can realize the naked eye detection of ONOO- due to the obvious color change. TPA-F-NO2 has the advantages of large stokes shift, high signal-to-noise ratio, high selectivity and sensitivity. The quantitative detection can be achieved in the range of 0-14 µM ONOO-. Due to its solvatochromic characteristics, TPA-F-NO2 has the potential to be used in OLEDs and other fields. In addition, 4-methylumbelliferone has a wide range of anticancer effects as an inhibitor of hyaluronic acid. We prepared TPA-MU-NPs by assembling TPA-F-NO2 and 4-methylumbelliferone. It also endows TPA-MU-NPs with ONOO- imaging function and anti-proliferation effect on breast cancer cells and other cells. This 'probe-drug' assembly strategy provides ideas for the design and optimization of dual-functional probes.


Sujet(s)
Colorants fluorescents , Acide peroxynitreux , Humains , Colorants fluorescents/toxicité , Colorants fluorescents/composition chimique , Acide peroxynitreux/composition chimique , Hymécromone , Dioxyde d'azote , Imagerie optique
13.
Cells ; 11(23)2022 Nov 25.
Article de Anglais | MEDLINE | ID: mdl-36497040

RÉSUMÉ

Radioresistant (RR) cells are poor prognostic factors for tumor recurrence and metastasis after radiotherapy. The hyaluronan (HA) synthesis inhibitor, 4-methylumbelliferone (4-MU), shows anti-tumor and anti-metastatic effects through suppressing HA synthase (HAS) expression in various cancer cells. We previously reported that the administration of 4-MU with X-ray irradiation enhanced radiosensitization. However, an effective sensitizer for radioresistant (RR) cells is yet to be established, and it is unknown whether 4-MU exerts radiosensitizing effects on RR cells. We investigated the radiosensitizing effects of 4-MU in RR cell models. This study revealed that 4-MU enhanced intracellular oxidative stress and suppressed the expression of cluster-of-differentiation (CD)-44 and cancer stem cell (CSC)-like phenotypes. Interestingly, eliminating extracellular HA using HA-degrading enzymes did not cause radiosensitization, whereas HAS3 knockdown using siRNA showed similar effects as 4-MU treatment. These results suggest that 4-MU treatment enhances radiosensitization of RR cells through enhancing oxidative stress and suppressing the CSC-like phenotype. Furthermore, the radiosensitizing mechanisms of 4-MU may involve HAS3 or intracellular HA synthesized by HAS3.


Sujet(s)
Hyaluronan synthases , Hymécromone , Tumeurs de la bouche , Radiosensibilisants , Carcinome épidermoïde de la tête et du cou , Humains , Lignée cellulaire tumorale , Mouvement cellulaire , Prolifération cellulaire , Hyaluronan synthases/génétique , Tumeurs de la bouche/radiothérapie , Récidive tumorale locale , Radiosensibilisants/pharmacologie , Carcinome épidermoïde de la tête et du cou/radiothérapie , Radiotolérance , Hymécromone/pharmacologie
14.
Analyst ; 147(20): 4489-4499, 2022 Oct 10.
Article de Anglais | MEDLINE | ID: mdl-36069248

RÉSUMÉ

A novel type of fluorescent and electrochemical dual-signal sensor was constructed for the sensitive and selective detection of iron ions (Fe3+) based on a fluorescent material (Chi-FITC-4MU), which was synthesized by combining the organic dye 4-methylumbelliferone (4-MU), chitosan, and fluorescein isothiocyanate (FITC) in a simple step process. The 4-MU could bind to Fe3+ to form a complex, and clearly improved the selectivity of Chi-FITC-4MU for Fe3+ detection. FITC showed excellent fluorescence performance and chitosan was beneficial to the curing of the material. By solidifying the fluorescent material on an ITO surface, the dual-signal detection of Fe3+ could be realized with excellent selectivity, stability, and anti-interference ability. Based on the unique fluorescence properties of this sensor, the concentration of Fe3+ could be visualized in the linear range of 0.1-100 µM based on the degree of fluorescence quenching. Moreover, the highly sensitive and rapid analysis of low concentrations of Fe3+ was achieved through the electrochemical properties of the ITO sensor. The limit of detection (LOD) and the corresponding linear range were 0.0184 nM and 0.1-500 nM, respectively. Furthermore, this dual-signal sensor was effectively used for the detection of Fe3+ in actual water.


Sujet(s)
Chitosane , Fer , Fluorescéine-5-isothiocyanate , Hymécromone , Ions , Fer/analyse , Spectrométrie de fluorescence , Eau
15.
Biol Reprod ; 107(4): 1097-1112, 2022 10 11.
Article de Anglais | MEDLINE | ID: mdl-35810327

RÉSUMÉ

Meiotic maturation and cumulus expansion are essential for the generation of a developmentally competent gamete, and both processes can be recapitulated in vitro. We used a closed time-lapse incubator (EmbryoScope+™) to establish morphokinetic parameters of meiotic progression and cumulus expansion in mice and correlated these outcomes with egg ploidy. The average time to germinal vesicle breakdown (GVBD), time to first polar body extrusion (PBE), and duration of meiosis I were 0.91 ± 0.01, 8.82 ± 0.06, and 7.93 ± 0.06 h, respectively. The overall rate of cumulus layer expansion was 0.091 ± 0.002 µm/min, and the velocity of expansion peaked during the first 8 h of in vitro maturation (IVM) and then slowed. IVM of oocytes exposed to Nocodazole, a microtubule disrupting agent, and cumulus oocyte complexes (COCs) to 4-methylumbelliferone, a hyaluronan synthesis inhibitor, resulted in a dose-dependent perturbation of morphokinetics, thereby validating the system. The incidence of euploidy following IVM was >90% for both denuded oocytes and intact COCs. No differences were observed between euploid and aneuploid eggs with respect to time to GVBD (0.90 ± 0.22 vs. 0.97 ± 0.19 h), time to PBE (8.89 ± 0.98 vs. 9.10 ± 1.42 h), duration of meiosis I (8.01 ± 0.91 vs. 8.13 ± 1.38 h), and overall rate and kinetics of cumulus expansion (0.089 ± 0.02 vs 0.088 ± 0.03 µm/min) (P > 0.05). These morphokinetic parameters provide novel quantitative and non-invasive metrics for the evaluation of meiotic maturation and cumulus expansion and will enable screening compounds that modulate these processes.


Sujet(s)
Cellules du cumulus , Techniques de maturation in vitro des ovocytes , Animaux , Cellules du cumulus/métabolisme , Femelle , Acide hyaluronique/métabolisme , Hymécromone/métabolisme , Techniques de maturation in vitro des ovocytes/méthodes , Méiose , Souris , Nocodazole , Ovocytes/métabolisme
16.
Am J Respir Cell Mol Biol ; 67(3): 360-374, 2022 09.
Article de Anglais | MEDLINE | ID: mdl-35679095

RÉSUMÉ

Allergic rhinitis (AR) is a multifactorial airway disease characterized by basal and goblet cell hyperplasia. Hyaluronic acid (HA) is a major component of extracellular matrix and a critical contributor to tissue repair and remodeling after injury. We previously demonstrated that the intermediate progenitor cell (IPC) surface marker CD44v3 is upregulated in the basal and suprabasal layers of well-differentiated primary human nasal epithelial (HNE) cells after stimulation with the Th2 (T-helper cell type 2) cytokine IL-4, and an antibody blocking the CD44v3-HA interaction suppressed IL-4-induced goblet cell hyperplasia. We now show that the expression of HA and two HA synthases, HAS2 and HAS3, was upregulated in both the nasal surface epithelium of subjects with AR and IL-4-stimulated HNE cells. Inhibition of HA synthesis by 4-methylumbelliferone suppressed IL-4-induced goblet cell hyperplasia. Moreover, HAS2 and HAS3 were expressed in IPCs depending on the differentiation events, as follows: the rapid, transient upregulation of HAS2 induced basal IPC proliferation and basal-to-suprabasal transition, whereas the delayed upregulation of HAS3 promoted the transition of suprabasal IPCs to a goblet cell fate. 4-methylumbelliferone treatment in a house dust mite-induced murine AR model attenuated goblet cell metaplasia. Last, HA concentrations in nasal epithelial lining fluids from patients with AR positively correlated with the concentrations of mediators causing allergic inflammation. These data suggest that HA produced after the sequential upregulation of HAS2 and HAS3 contributes to goblet cell hyperplasia in allergic airway inflammation and modulates disease progression.


Sujet(s)
Cellules caliciformes , Hyaluronan synthases , Rhinite allergique , Animaux , Cellules caliciformes/effets des médicaments et des substances chimiques , Cellules caliciformes/enzymologie , Cellules caliciformes/anatomopathologie , Humains , Hyaluronan synthases/métabolisme , Acide hyaluronique/métabolisme , Hymécromone/pharmacologie , Hymécromone/usage thérapeutique , Hyperplasie/génétique , Hyperplasie/anatomopathologie , Interleukine-4/métabolisme , Souris , Rhinite allergique/traitement médicamenteux , Rhinite allergique/enzymologie , Rhinite allergique/anatomopathologie
17.
J Clin Invest ; 132(9)2022 05 02.
Article de Anglais | MEDLINE | ID: mdl-35499083

RÉSUMÉ

BACKGROUNDHyaluronan (HA), an extracellular matrix glycosaminoglycan, has been implicated in the pathophysiology of COVID-19 infection, pulmonary hypertension, pulmonary fibrosis, and other diseases, but is not targeted by any approved drugs. We asked whether hymecromone (4-methylumbelliferone [4-MU]), an oral drug approved in Europe for biliary spasm treatment that also inhibits HA in vitro and in animal models, could be repurposed as an inhibitor of HA synthesis in humans.METHODSWe conducted an open-label, single-center, dose-response study of hymecromone in healthy adults. Subjects received hymecromone at 1200 (n = 8), 2400 (n = 9), or 3600 (n = 9) mg/d divided into 3 doses daily, administered orally for 4 days. We assessed safety and tolerability of hymecromone and analyzed HA, 4-MU, and 4-methylumbelliferyl glucuronide (4-MUG; the main metabolite of 4-MU) concentrations in sputum and serum.RESULTSHymecromone was well tolerated up to doses of 3600 mg/d. Both sputum and serum drug concentrations increased in a dose-dependent manner, indicating that higher doses lead to greater exposures. Across all dose arms combined, we observed a significant decrease in sputum HA from baseline after 4 days of treatment. We also observed a decrease in serum HA. Additionally, higher baseline sputum HA levels were associated with a greater decrease in sputum HA.CONCLUSIONAfter 4 days of exposure to oral hymecromone, healthy human subjects experienced a significant reduction in sputum HA levels, indicating this oral therapy may have potential in pulmonary diseases where HA is implicated in pathogenesis.TRIAL REGISTRATIONClinicalTrials.gov NCT02780752.FUNDINGStanford Medicine Catalyst, Stanford SPARK, Stanford Innovative Medicines Accelerator program, NIH training grants 5T32AI052073-14 and T32HL129970.


Sujet(s)
Acide hyaluronique , Hymécromone , Administration par voie orale , COVID-19 , Europe , Matrice extracellulaire/métabolisme , Humains , Acide hyaluronique/métabolisme , Hymécromone/administration et posologie , Hymécromone/effets indésirables
18.
J Mater Chem B ; 10(20): 3916-3926, 2022 05 25.
Article de Anglais | MEDLINE | ID: mdl-35485215

RÉSUMÉ

Glucuronidation is a metabolic pathway that inactivates many drugs including hymecromone. Adverse effects of glucuronide metabolites include a reduction of half-life circulation times and rapid elimination from the body. Herein, we developed synthetic catalytic nanocompartments able to cleave the glucuronide moiety from the metabolized form of hymecromone in order to convert it to the active drug. By shielding enzymes from their surroundings, catalytic nanocompartments favor prolonged activity and lower immunogenicity as key aspects to improve the therapeutic solution. The catalytic nanocompartments (CNCs) consist of self-assembled poly(dimethylsiloxane)-block-poly(2-methyl-2-oxazoline) diblock copolymer polymersomes encapsulating ß-glucuronidase. Insertion of melittin in the synthetic membrane of these polymersomes provided pores for the diffusion of the hydrophilic hymecromone-glucuronide conjugate to the compartment inside where the encapsulated ß-glucuronidase catalyzed its conversion to hymecromone. Our system successfully produced hymecromone from its glucuronide conjugate in both phosphate buffered solution and cell culture medium. CNCs were non-cytotoxic when incubated with HepG2 cells. After being taken up by cells, CNCs produced the drug in situ over 24 hours. Such catalytic platforms, which locally revert a drug metabolite into its active form, open new avenues in the design of therapeutics that aim at prolonging the residence time of a drug.


Sujet(s)
Glucuronides , Hymécromone , Catalyse , Glucuronidase/métabolisme , Glucuronides/métabolisme , Hymécromone/métabolisme , Polymères
19.
Int J Mol Sci ; 23(5)2022 Feb 22.
Article de Anglais | MEDLINE | ID: mdl-35269567

RÉSUMÉ

Hyaluronan (HA), an essential component of the extracellular matrix of the skin, is synthesized by HA synthases (HAS1-3). To date, epidermal HA has been considered a major player in regulating cell proliferation and differentiation. However, a previous study reported that depletion of epidermal HA by Streptomyces hyaluronidase (St-HAase) has no influence on epidermal structure and function. In the present study, to further explore roles of epidermal HA, we examined effects of siRNA-mediated knockdown of HAS3, as well as conventional HA-depletion methods using St-HAase and 4-methylumbelliferone (4MU), on epidermal turnover and architecture in reconstructed skin or epidermal equivalents. Consistent with previous findings, HA depletion by St-HAase did not have a substantial influence on the epidermal architecture and turnover in skin equivalents. 4MU treatment resulted in reduced keratinocyte proliferation and epidermal thinning but did not seem to substantially decrease the abundance of extracellular HA. In contrast, siRNA-mediated knockdown of HAS3 in epidermal equivalents resulted in a significant reduction in epidermal HA content and thickness, accompanied by decreased keratinocyte proliferation and differentiation. These results suggest that HAS3-mediated HA production, rather than extracellularly deposited HA, may play a role in keratinocyte proliferation and differentiation, at least in the developing epidermis in reconstructed epidermal equivalents.


Sujet(s)
Hyaluronan synthases/génétique , Acide hyaluronique/métabolisme , Hyaluronoglucosaminidase/pharmacologie , Hymécromone/pharmacologie , Kératinocytes/cytologie , Protéines bactériennes/pharmacologie , Différenciation cellulaire/effets des médicaments et des substances chimiques , Prolifération cellulaire/effets des médicaments et des substances chimiques , Cellules cultivées , Épiderme/effets des médicaments et des substances chimiques , Épiderme/métabolisme , Régulation de l'expression des gènes/effets des médicaments et des substances chimiques , Techniques de knock-down de gènes , Humains , Kératinocytes/effets des médicaments et des substances chimiques , Kératinocytes/métabolisme , Streptomyces/enzymologie
20.
Signal Transduct Target Ther ; 7(1): 91, 2022 03 18.
Article de Anglais | MEDLINE | ID: mdl-35304437

RÉSUMÉ

Currently, there is no effective drugs for treating clinically COVID-19 except dexamethasone. We previously revealed that human identical sequences of SARS-CoV-2 promote the COVID-19 progression by upregulating hyaluronic acid (HA). As the inhibitor of HA synthesis, hymecromone is an approved prescription drug used for treating biliary spasm. Here, we aimed to investigate the relation between HA and COVID-19, and evaluate the therapeutic effects of hymecromone on COVID-19. Firstly, HA was closely relevant to clinical parameters, including lymphocytes (n = 158; r = -0.50; P < 0.0001), C-reactive protein (n = 156; r = 0.55; P < 0.0001), D-dimer (n = 154; r = 0.38; P < 0.0001), and fibrinogen (n = 152; r = 0.37; P < 0.0001), as well as the mass (n = 78; r = 0.43; P < 0.0001) and volume (n = 78; r = 0.41; P = 0.0002) of ground-glass opacity, the mass (n = 78; r = 0.48; P < 0.0001) and volume (n = 78; r = 0.47; P < 0.0001) of consolidation in patient with low level of hyaluronan (HA < 48.43 ng/mL). Furthermore, hyaluronan could directly cause mouse pulmonary lesions. Besides, hymecromone remarkably reduced HA via downregulating HAS2/HAS3 expression. Moreover, 89% patients with hymecromone treatment had pulmonary lesion absorption while only 42% patients in control group had pulmonary lesion absorption (P < 0.0001). In addition, lymphocytes recovered more quickly in hymecromone-treated patients (n = 8) than control group (n = 5) (P < 0.05). These findings suggest that hymecromone is a promising drug for COVID-19 and deserves our further efforts to determine its effect in a larger cohort.


Sujet(s)
Traitements médicamenteux de la COVID-19 , Acide hyaluronique , Animaux , Humains , Hymécromone/métabolisme , Hymécromone/pharmacologie , Souris , Ordonnances , SARS-CoV-2
SÉLECTION CITATIONS
DÉTAIL DE RECHERCHE
...