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1.
Chem Commun (Camb) ; 59(44): 6609-6626, 2023 May 30.
Artículo en Inglés | MEDLINE | ID: mdl-37161668

RESUMEN

Extracellular vesicles (EVs) are nanosized vesicles enclosed in a lipid membrane that are sustainably released by nearly all cell types. EVs have been deemed as valuable biomarkers for diagnostics and effective drug carriers, owing to the physiological function of transporting biomolecules for intercellular communication. To investigate their biological properties, efficient labeling strategies have been constructed for EV research, among which fluorescence labeling exerts a powerful function due to the capability of visualizing the nanovesicles with high sensitivity both in vitro and in vivo. In one aspect, with the help of functional fluorescence tags, EVs could be differentiated and categorized in vitro by various analytical techniques, which exert vital roles in disease diagnosis, prognosis, and treatment monitoring. Additionally, innovative EV reporters have been utilized for visualizing EVs, in combination with powerful microscopy techniques, which provide potential tools for investigating the dynamic events of EV release and intercellular communication in suitable animal models. In this feature article, we survey the latest advances regarding EV fluorescence labeling strategies and their application in biomedical application and in vivo biology investigation, highlighting the progresses in individual EV imaging. Finally, the challenges and future perspectives in unravelling EV physiological properties and further biomedical application are discussed.


Asunto(s)
Vesículas Extracelulares , Lípidos/química , Colorantes Fluorescentes/química , Vesículas Extracelulares/química , Humanos , Animales , Citometría de Flujo , Microscopía Fluorescente , Comunicación Celular , Transporte Biológico
2.
Artículo en Inglés | MEDLINE | ID: mdl-36636608

RESUMEN

Background: Clinacanthus nutans (Burm.f.) Lindau (C. nutans) has been used in the therapy of hepatitis B (HB) and is effective; however, the mechanism of action has not been elucidated. Objective: To investigate the protective effects of C. nutans aqueous extract on the hepatitis B virus (HBV) mouse model based on correlation analysis between gut microbiota and liver metabolomics. Materials and Methods: We firstly constructed the animal model by high-pressure injection of pcDNA3.1(+)/HBV plasmid into the tail vein and treated it with C. nutans. The biomarkers and inflammatory cytokines of HB were detected by enzyme-linked immunosorbent assay and quantitative PCR; the Illumina-MiSeq platform was used for investigating gut microbiota; the LC-MS/MS method was utilized on screening liver tissue metabolites; multiomics joint analysis was performed using the R program. Results: Compared with the modeling group, C. nutans significantly decreased the expression levels of HBsAg, IL-1ß, TNF-α(P < 0.05) in the serum, and cccDNA (P < 0.05) in the liver tissues of mice. C. nutans dramatically reduced the ratio of Firmicutes and Bacteroidetes (P < 0.05) and significantly declined the proportion of Lactobacillaceae and Lactobacillus(P < 0.05), dramatically increasing the relative abundance of Bacteroidales_S24-7_group, Rikenellaceae, and Alistipes(P < 0.05); LC-MS/MS analysis results showed that C. nutans dramatically upregulate hippuric acid, L-histidine, trehalose, D-threitol, and stachyose and downregulate uridine 5'-diphosphate, cholic acid, trimethylamine N-oxide, CDP-ethanolamine, and phosphorylcholine (P < 0.05). The correlation analysis revealed that C. nutans affects the related metabolite levels of hippuric acid and cholic acid through the modulation of crucial bacteria (Alistipes) (P < 0.01), exerting specific anti-inflammatory effects. Conclusion: These results suggest that C. nutans exerts protective effects in HBV model mice, showing the therapeutic potential for anti-HBV infection.

3.
J Diabetes Res ; 2018: 4230590, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-29967794

RESUMEN

Recently, the role of gut microbiota in the development of obesity and type 2 diabetes mellitus (T2DM) has been highlighted. We performed an 8-week administration protocol on T2DM (C57BL/6J db-/db-) mice and fecal samples were collected. Comparisons of fecal bacterial communities were performed between db-/db- mice and normal mice (DB/DB) and between the db-/db mice treated and untreated with AOE using next-generation sequencing technology. Our results showed that the db-/db-AOE group had improved glycemic control and renal function compared with the db-/db-H2O group. Compared with the db-/db-H2O group, AOE administration resulted in significantly increased ratio of Bacteroidetes-to-Firmicutes in db-/db- mice. In addition, the abundance of Akkermansia was significantly increased, while Helicobacter was significantly suppressed in the db-/db-AOE group compared with the db-/db-H2O group. Our data suggest that AOE treatment decreased blood glucose levels and significantly reduced damage of renal pathology in the T2DM mice by modulating gut microbiota composition.


Asunto(s)
Alpinia , Diabetes Mellitus Tipo 2/prevención & control , Microbioma Gastrointestinal/efectos de los fármacos , Extractos Vegetales/uso terapéutico , Animales , Glucemia/metabolismo , Diabetes Mellitus Tipo 2/sangre , Diabetes Mellitus Tipo 2/microbiología , Heces/microbiología , Tracto Gastrointestinal/microbiología , Prueba de Tolerancia a la Glucosa , Masculino , Ratones , Extractos Vegetales/administración & dosificación
4.
Oncol Lett ; 15(4): 4797-4804, 2018 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-29552119

RESUMEN

Programmed cell death-1 (PD-1) is an oncogene associated with suppressing proliferation and cytokine production of T cells in the progression of liver cancer. microRNAs (miRs) regulate gene expression via specific binding to the target 3'untranslated region of mRNA. In the present study, miR-374b was indicated to interact with PD-1 and affect the tumor-targeting capacity of cytokine-induced killer (CIK) cells. miR-374b inhibitor significantly increased PD-1 expression in CIK cells. A synthetic small interfering (si)RNA targeting PD-1 was employed to silence the expression level of PD-1 in CIK cells. Then, the antitumor effect of siPD-1 in CIK cells was investigated. In vitro study demonstrated that IFN-γ secretion and the concentration of lactate dehydrogenase were significantly increased in the PD-1 knockdown group; however, the viability of HepG2 cells in the PD-1 knockdown group had significantly decreased, compared with the HepG2 cells in the negative control group. In vivo study indicated that mice inoculated with HepG2 cells and CIK cells with PD-1 knocked down had a significantly smaller tumor volume, compared with the control group. To conclude, human CIK cells transfected with siPD-1 can target liver cancer cells and enhance immunotherapy efficacy, and therefore they have potential in the immunotherapy of liver cancer.

5.
Exp Ther Med ; 13(4): 1321-1328, 2017 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-28413472

RESUMEN

Diabetes mellitus is characterized by high blood glucose levels. Increased levels of reactive oxygen species (ROS) may disrupt insulin signaling and result in insulin resistance. The Alpinia oxyphylla extract (AOE) possesses powerful antioxidant activity and may therefore inhibit the development of insulin resistance. The objective of the present study was to determine the effects of AOE on blood glucose, insulin and lipid levels in a type II diabetic nephropathy animal model (C57BIKsj db-/db-). All experiments were performed on male C57BL/Ks DB/DB and db-/db- mice that were left to acclimatize for 1 week prior to the experimental period. AOE was administered to these mice at different dosages (100, 300 and 500 mg/kg) for 8 weeks. The results demonstrated that AOE did not affect mouse weight, while blood glucose concentrations were found to significantly decrease in a dose-dependent manner (P<0.05). The effect of administering 500 mg/kg AOE (AOE500) to db-/db- mice was tested further. Treatment with AOE500 for 8 weeks led to improved glucose tolerance and reduced plasma insulin concentrations (P<0.05), as well as a significant decrease in triglyceride concentrations (P<0.05) and levels of total cholesterol (P<0.05) in db-/db- mice. Furthermore, treatment with AOE500 decreased the concentration of malondialdehyde, elevated the concentration of glutathione and increased the activities of the antioxidant enzymes superoxide dismutase and peroxidase (P<0.05) in the livers of db-/db- mice. Meanwhile, AOE-treated mice exhibited significantly reduced urine albumin, creatinine and blood urea nitrogen excretion (P<0.05). In parallel, the upregulated expression of phosphatase and tensin homolog (PTEN) in the liver and kidneys of db-/db- mice was impaired following AOE500 treatment. The results of the present study suggest that AOE regulates blood glucose and lipid levels and improves renal function by mediating oxidative stress and PTEN expression at the onset of type II diabetes mellitus.

6.
Biol Res ; 50(1): 9, 2017 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-28249617

RESUMEN

BACKGROUND: A number of dysregulated miRNAs have been identified and are proposed to have significant roles in the pathogenesis of type 2 diabetes mellitus or renal pathology. Alpinia oxyphylla has shown significant anti-inflammatory properties and play an anti-diabetes role. The objective of this study was to detect the alteration of miRNAs underlying the anti-diabetes effects of A. oxyphylla extract (AOE) in a type II diabetic animal model (C57BIKsj db-/db-). RESULTS: Treatment with AOE for 8 weeks led to lower concentrations of blood glucose, urine albumin, and urine creatinine. 17 and 13 miRNAs were statistically identified as differentially regulated in the DB/DB and db-/db- AOE mice, respectively, compared to the untreated db-/db- mice. Of these, 7 miRNAs were identified in both comparison groups, and these 7 miRNAs were verified by quantitative real-time PCR. Functional bioinformatics showed that the putative target genes of 7 miRNAs were associated with several diabetes effects and signaling pathways. CONCLUSIONS: These founding suggest that the potential of AOE as a medicinal anti-diabetes treatment through changes in the expressions of specific miRNAs. The results provide a useful resource for future investigation of the role of AOE-regulated miRNAs in diabetes mellitus.


Asunto(s)
Diabetes Mellitus Tipo 2/tratamiento farmacológico , Hipoglucemiantes/farmacología , Riñón/efectos de los fármacos , MicroARNs/efectos de los fármacos , Extractos Vegetales/farmacología , Albuminuria , Alpinia , Animales , Glucemia/análisis , Creatinina/sangre , Diabetes Mellitus Experimental/tratamiento farmacológico , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Nefropatías Diabéticas/tratamiento farmacológico , Nefropatías Diabéticas/metabolismo , Regulación de la Expresión Génica , Riñón/metabolismo , Masculino , Ratones Endogámicos C57BL , MicroARNs/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Reproducibilidad de los Resultados , Análisis de Secuencia de ARN , Factores de Tiempo , Resultado del Tratamiento
7.
Biol. Res ; 50: 9, 2017. tab, graf
Artículo en Inglés | LILACS | ID: biblio-838964

RESUMEN

BACKGROUND: A number of dysregulated miRNAs have been identified and are proposed to have significant roles in the pathogenesis of type 2 diabetes mellitus or renal pathology. Alpinia oxyphylla has shown significant anti-inflammatory properties and play an anti-diabetes role. The objective of this study was to detect the alteration of miRNAs underlying the anti-diabetes effects of A. oxyphylla extract (AOE) in a type II diabetic animal model (C57BIKsj db-/db-). RESULTS: Treatment with AOE for 8 weeks led to lower concentrations of blood glucose, urine albumin, and urine creatinine. 17 and 13 miRNAs were statistically identified as differentially regulated in the DB/DB and db-/db- AOE mice, respectively, compared to the untreated db-/db- mice. Of these, 7 miRNAs were identified in both comparison groups, and these 7 miRNAs were verified by quantitative real-time PCR. Functional bioinformatics showed that the putative target genes of 7 miRNAs were associated with several diabetes effects and signaling pathways. CONCLUSIONS: These founding suggest that the potential of AOE as a medicinal anti-diabetes treatment through changes in the expressions of specific miRNAs. The results provide a useful resource for future investigation of the role of AOE-regulated miRNAs in diabetes mellitus.


Asunto(s)
Animales , Masculino , Ratones , Extractos Vegetales/farmacología , MicroARNs/efectos de los fármacos , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Hipoglucemiantes/farmacología , Riñón/efectos de los fármacos , Factores de Tiempo , Glucemia/análisis , Regulación de la Expresión Génica , Reproducibilidad de los Resultados , Resultado del Tratamiento , Análisis de Secuencia de ARN , Creatinina/sangre , MicroARNs/metabolismo , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Experimental/tratamiento farmacológico , Diabetes Mellitus Tipo 2/metabolismo , Nefropatías Diabéticas/metabolismo , Nefropatías Diabéticas/tratamiento farmacológico , Albuminuria , Reacción en Cadena en Tiempo Real de la Polimerasa , Riñón/metabolismo , Ratones Endogámicos C57BL
8.
Int J Clin Exp Med ; 7(2): 379-83, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24600492

RESUMEN

The purpose of this study was to investigate the influencing mechanism of Porphyromonas gingivalis extracellular vesicles on human periodontal ligament fibroblasts to better understand the pathogenesis of periodontitis, the major cause of adult tooth loss. Human periodontal ligament fibroblasts were cultured and randomly assigned to a control group and an extracellular vesicles (ECV) group. The ECV group was exposed to isolated Porphyromonas gingivalis extracellular vesicles; the control group was not exposed. Western blotting was used to detect expression of matrix metalloproteinase 1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1), and RT-PCR was used to detect mRNA expression of alkaline phosphatase (ALP). When human periodontal ligament fibroblasts were processed by Porphyromonas gingivalis extracellular vesicles (ECV), protein expression levels of both MMP-1 and TIMP-1 were significantly higher than that of the control group (P<0.05). In contrast, ALP mRNA expression in human periodontal ligament fibroblasts processed by ECV was significantly lower than that of the control group (P<0.05). Porphyromonas gingivalis extracellular vesicles can up-regulate expression of MMP-1 and TIMP-1 protein and ALP mRNA of human periodontal fibroblasts.

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