Carbohydrate Polymer Technologies and Applications
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2025
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IF 6.5
Hurong Ge,Peipei Ma,Qing Huang,Shijie Wei,Zhizhong Wang
Objectives: To study the encapsulation of resveratrol by three cyclodextrin derivatives, investigate the impact of encapsulation technology on its anti-fatigue effect, and explore the anti-fatigue mechanism of the encapsulated resveratrol via transcriptomics analysis of exosomal miRNAs. Methods (1) UV spectroscopy and nuclear magnetic resonance (NMR) were used to investigate the encapsulation and encapsulation modes of resveratrol by β -CD, HP- β -CD and CM- β -CD; and the phase solubility method was used to calculate the solubilisation multiplicity of resveratrol by the three cyclodextrin derivatives. (2) Male C57BL/6 J mice were randomly divided into: fatigue model group (FM), FM+resveratrol group (SF+Res), FM+ β -CD, FM+HP- β -CD, FM+CM- β -CD, FM+ β -CD+Res, FM+HP- β -CD+Res, FM+CM- β -CD +Res, and after 30 days of rearing, the mice were subjected to exhaustion swimming experiment to detect fatigue-related indexes and observe the effects of the three cyclodextrin derivatives on the anti-fatigue effects of resveratrol. (3) Male C57BL/6 J mice were randomly divided into Control, FM, and FM+HP- β -CD+Res. After 30 days of rearing, except for the Control group, the remaining mice were subjected to exhaustion swimming experiments, and exosomes were extracted from cardiac tissue, skeletal muscle tissue, and liver tissue of the three groups of mice, and the transcriptomics technology was applied to sequence the exosomal miRNAs and to explore the mechanism of the effect of the HP- β -CD+Res envelope on fatigue. Results (1) The inclusion ratios of β -CD, HP- β -CD, and CM- β -CD encapsulated resveratrol were 1:1, 1:2, and 1:1, and the solubilisation multiplicities of resveratrol were 23, 39, and 21 fold, respectively; β -CD and CM- β -CD encapsulated Res, which was the benzene ring with two hydroxyl‑terminal ends of Res entered the cavity from the small-ported end of the cyclodextrins; HP- β -CD encapsulated Res, which was the Res containing one hydroxyl‑terminated benzene ring enters the cavity from the wide-mouth end of HP-β-CD. (2) Compared with the FM+Res group, all three inclusions were able to prolong the exhaustion swimming time of mice, with HP- β -CD+Res being the most effective. (3) After HP- β -CD+Res acted on the fatigued mouse model, cardiac tissue showed 1726 differentially expressed exosomal miRNAs, of which 840 were up-regulated and 886 were down-regulated, reversing the fatigue-induced changes in 64.96 % of the exosomal miRNAs (1170 shared differential exosomal miRNAs); Skeletal muscle tissue showed 1948 differentially expressed exosomal miRNAs, of which 718 were up-regulated and 1230 were down-regulated, reversing 80.57 % of the fatigue-induced changes in exosomal miRNAs (1576 shared differential exosomal miRNAs); Liver tissue showed 1442 differentially expressed exosomal miRNAs, of which 451 were up-regulated and 991 were down-regulated, reversing 84.31 % of fatigue-induced changes in exosomal miRNAs (1214 shared differential exosomal miRNAs). Target gene enrichment analysis of differentially expressed exosomal miRNAs in cardiac tissue, skeletal muscle tissue, and liver tissue had similar results, and the biological processes involved in HP- β -CD+Res were related to biological_process, signal transduction, G protein-coupled receptor signaling pathway, etc., and cellular components such as membrane, cytoplasm, integral component of membrane, etc., and molecular functions are closely related to protein binding, metal ion binding, molecular_function, etc.; mainly enriched in Axon guidance, MicroRNAs in cancer, Apoptosis and other signalling pathways. Conclusions (1) All three cyclodextrin derivatives encapsulated increased the solubility of resveratrol, with HP- β -CD having the best effect. (2) All three cyclodextrin derivative encapsulation techniques improved the anti-fatigue effect of resveratrol, with HP- β -CD being the most effective. (3) HP- β -CD+Res was able to induce differential expression of exosomal miRNAs in cardiac tissue, skeletal muscle tissue and liver tissue of fatigued mice.
研究领域:
药理学
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分子生物学
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生物信息学
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产品关联
DOI
高影响因子
STAR Protocols
·
2023
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IF 0
Kun Li,Chunzi Liang,Yuling Pan,Linghan Mei,Guo-Jun Zhang
Summary Here, we present a protocol for constructing an ultrasensitive biosensor for exosomal-miRNA detection. We describe steps for preparing graphene quantum dot-phosphorodiamidate morpholino oligomer hybrids, depositing them onto the reduced graphene oxide field surface, hybridizing analyte miRNA with the sensor probe, and capturing and calculating electrical signals. We also detail procedures for optimizing biosensor construction and evaluating performance. By quantifying plasma exosomal miRNA21, this protocol can identify cancer patients from healthy individuals. For complete details on the use and execution of this protocol, please refer to Li et al. 1
研究领域:
纳米技术
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生物传感器
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分子生物学
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癌症研究
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DOI
PubMed
Frontiers in Genetics
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2021
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IF 4.599
Zhang Jing,Gao Juan,Lin Dengna,Xiong Jing,Wang Jialei,Chen Junfeng,Lin Bingliang,Gao Zhiliang
Acute-on-chronic liver failure (ACLF) is a severe syndrome associated with high mortality. Alterations in the liver microenvironment are one of the vital causes of immune damage and liver dysfunction. Human bone marrow mesenchymal stem cells (hBMSCs) have been reported to alleviate liver injury via exosome-mediated signaling; of note, miRNAs are one of the most important cargoes in exosomes. Importantly, the miRNAs within exosomes in the hepatic microenvironment may mediate the mesenchymal stem cell (MSC)-derived regulation of liver function. This study investigated the hepatocyte exosomal miRNAs which are regulated by MSCs and the target genes which have potential in the treatment of liver failure. Briefly, ACLF was induced in mice using carbon tetrachloride and primary hepatocytes were isolated and co-cultured (or not) with MSCs under serum-free conditions. Exosomes were then collected, and the expression of exosomal miRNAs was assessed using next-generation sequencing; a comparison was performed between liver cells from healthy versus ACLF animals. Additionally, to identify the intracellular targets of exosomal miRNAs in humans, we focused on previously published data, i.e., microarray data and mass spectrometry data in liver samples from ACLF patients. The biological functions and signaling pathways associated with differentially expressed genes were predicted using gene ontology and Kyoto Encyclopedia of Genes and Genomics enrichment analyses; hub genes were also screened based on pathway analysis and the prediction of protein-protein interaction networks. Finally, we constructed the hub gene-miRNA network and performed correlation analysis and qPCR validation. Importantly, our data revealed that MSCs could regulate the miRNA content within exosomes in the hepatic microenvironment. MiR-20a-5p was down-regulated in ACLF hepatocytes and their exosomes, while the levels of chemokine C-X-C Motif Chemokine Ligand 8 (CXCL8; interleukin 8) were increased in hepatocytes. Importantly, co-culture with hBMSCs resulted in up-regulated expression of miR-20a-5p in exosomes and hepatocytes, and down-regulated expression of CXCL8 in hepatocytes. Altogether, our data suggest that the exosomal miR-20a-5p/intracellular CXCL8 axis may play an important role in the reduction of liver inflammation in ACLF in the context of MSC-based therapies and highlights CXCL8 as a potential target for alleviating liver injury.
研究领域:
肝病学
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干细胞生物学
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分子生物学
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基因组学
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产品关联
DOI
PubMed
高影响因子
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS
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2024
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IF 3.8
Ying Zhang,Jing Xie,Huimin Zhang,Jiacheng Li,Xing Mi,Xuyi Zhou,Zhenhua Ding
Introduction Local exposure to ionizing radiation (IR) can induce changes in biological processes in distant tissues and organs. Exosomes are nanoscale vesicles that transport biomolecules, mediate communication between cells and tissues, and can affect the abscopal effects of radiotherapy. Methods Mice were treated with 8.0 Gy doses of chest and abdomen IR, after which serum samples were taken 24 h after exposure. Their serum exosomes were then isolated via ultracentrifugation and the small RNA portions were extracted for sequencing and bioinformatic analysis. Exosomes were injected intravenously into the mice to assess their ability to cross the blood-brain barrier (BBB). Glioma cells and gliomal stem cells (GSCs) were examined for malignant biological behaviors, stemness, and tumorigenic capacity after co-culturing with different types of exosomes. Results We found that serum exosomes crossed the BBB in mice after local IR exposure—which induced decreases in the expression of BBB tight-junction proteins and increased brain endothelial cell apoptosis. Exosomes from the exposed groups promoted malignant biological behaviors, stemness, and tumorigenic capacity in glioma cells and GSCs by upregulating the expression of SOS1. Phospho-MEK1/2 and Phospho-ERK1/2, of the MAPK signaling pathway, were found to be up-regulated in cells that were co-cultured with the exposing groups of the exosomes. Further analyses demonstrated that differentially expressed levels of miR-93-5p in mouse serum exosomes regulated the cellular expression of SOS1. Conclusion Following local IR exposure, serum exosomes cross the BBB to promote the progression of distant gliomas. Exosomal microRNAs play an important role in this process.
研究领域:
肿瘤学
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分子生物学
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放射生物学
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产品关联
DOI
PubMed
高影响因子
Oxidative Medicine and Cellular Longevity
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2019
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IF 4.868
Lifang Sun,Min Zhu,Wei Feng,Yiping Lin,Jia Yin,Juan Jin,Yunguang Wang
Idiopathic pulmonary fibrosis (IPF) is a prototype of chronic, progressive, and fibrotic lung disease with high morbidity and high mortality. Menstrual blood-derived stem cells (MenSCs) have proven to be an attractive tool for the treatment of acute lung injury and fibrosis-related diseases through immunosuppression and antifibrosis. However, whether MenSC-derived exosomes have the similar function on pulmonary fibrosis remains unclear. In the present study, exosomes secreted from MenSCs (MenSCs-Exo) were verified by transmission electron microscope (TEM), nanoparticle tracking analyzer (NTA), and western blotting. And MenSC-Exo addition significantly improved BLM-induced lung fibrosis and alveolar epithelial cell damage in mice, mainly reflected in BLM-mediated enhancement of the fibrosis score, blue collagen deposition, dry/wet gravity ratio, hydroxyproline and malondialdehyde levels, and downregulation of glutathione peroxidase, which were all robustly reversed by MenSC-Exo management. Additionally, BLM- and TGF-β1-evoked cellular reactive oxygen species (ROS), mitochondrial DNA (mtDNA) damage, and cell apoptosis were rescued by MenSCs-Exo in vivo and in vitro. Further study indicated that the MenSCs-Exo could transport miRNA Let-7 into recipient alveolar epithelial cells. Let-7 inhibitor administration significantly blocked the exosome-mediated improvement role on lung fibrosis in mice. Mechanistically, Let-7 was able to regulate the expression of lectin-like oxidized low-density lipoprotein receptor-1 (LOX1) through binding to its 3-UTR region. Forced expression of LOX1 promoted the expression of apoptosis-related protein and mtDNA damage markers via regulating NLRP3 which was also confirmed in BLM model mice under the combination therapy of the exosome and Let-7 inhibitor. Collectively, this study demonstrates that exosomal Let-7 from MenSCs remits pulmonary fibrosis through regulating ROS, mtDNA damage, and NLRP3 inflammasome activation. This provides a new approach of exocytosis on the treatment of fibrotic lung disease.
研究领域:
干细胞生物学
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肺医学
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分子生物学
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细胞生物学
✓ 已验证
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2024
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IF 6.7
Guohua Qi,Xingkang Diao,Yu Tian,Dan Sun,Yongdong Jin
Exosomal miRNA expression derived from tumor cells provides a valuable and promising noninvasive modality for the early diagnosis and assessment of the efficacy of cancer treatment. However, accurate detection and identification of miRNA within exosomes have been challenging due to its low abundance and the complexity and tedious extraction with large sample volumes in the separation process. Here, we developed an electrically activated nanoplatform for rapid and sensitive detection and identification of exosome miRNA, through triggering miRNA release by opening exosomes that were captured on the electrode surface using a slightly applied electric field (50 mV), and simultaneously detected them with surface-enhanced Raman spectroscopy (SERS) in situ. The method possessed superior specificity and sensitivity for exosomal miRNA detection, with a low detection concentration of 0.5 nM. The SERS sensor chips also showed a superior sensing performance of exosomal miRNA in complex body fluids such as urine and blood. We found that exosomal miRNA contents derived from tumor cells were significantly higher than those in normal cells, and importantly, the concentrations of exosomes secreted from three different cell lines were distinctly augmented after mild electrical stimulation (ES) treatment. Furthermore, the miRNA expression within exosomes was upregulated after the ES treatment of cells. The developed approach and SERS detection platform for exosomal miRNA are promising for noninvasive and precise screening, classification, and monitoring of cancer.
研究领域:
癌症生物学
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纳米技术
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生物医学工程
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分析化学
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分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
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