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1.
Sci Rep ; 11(1): 8789, 2021 04 22.
Article in English | MEDLINE | ID: mdl-33888814

ABSTRACT

To explore the pathogenesis of high myopia (HM) using quantitative proteomics. The aqueous humor of patients with simple nuclear cataract and nuclear cataract complicated with HM (hereinafter referred to as "C" and "HM" groups, respectively) were collected. The isobaric tags for relative and absolute quantitation (iTRAQ)-based liquid chromatography-tandem mass spectrometry (LC-MS/MS) proteomics approach was employed to explore differentially expressed proteins (DEPs). Bioinformatics was used to interpret the proteomic results. Furthermore, the plasminogen (PLG) protein was confirmed by enzyme-linked immunosorbent assay (ELISA) as the candidate biomarker for HM through a receiver operating characteristic curve analysis. The study showed 32 upregulated and 26 downregulated proteins. The gene ontology analysis demonstrated that 58 DEPs corresponded to 325 biological processes, 33 cell components, and 45 molecular functional annotations. The Kyoto Encyclopedia of Genes and Genomes analysis showed that the upregulated DEPs were highly enriched in the coagulation and complement cascades, consistent with the gene set enrichment analysis. Our data suggested that some DEPs might be hallmarks of the development of HM. ELISA confirmed that the PLG expression levels were significantly upregulated in HM. This was a new study investigating alterations in protein levels and affected pathways in HM using iTRAQ-based quantitative proteomics. Our study provided a comprehensive dataset on overall protein changes and shed light on its potential molecular mechanism in human HM.


Subject(s)
Aqueous Humor/metabolism , Myopia/metabolism , Plasminogen/metabolism , Proteomics/methods , Aged , Biomarkers/metabolism , Chromatography, Liquid/methods , Enzyme-Linked Immunosorbent Assay/methods , Female , Humans , Male , Middle Aged , Tandem Mass Spectrometry/methods
2.
Curr Eye Res ; 44(10): 1112-1120, 2019 10.
Article in English | MEDLINE | ID: mdl-31188648

ABSTRACT

Purpose: To observe the protective effects and underlying mechanisms of dl-3-n-butylphthalide (NBP) against H2O2-induced oxidative damage in retinal Müller cells. Methods: Cultured human Müller cell line (MIO-M1line) were exposed to H2O2 for 2 hours. Cell survival was evaluated by Calcein AM cell viability assay. Dichlorofluorescein diacetate (DCFDA) + endoplasmic reticulum (ER) red fluorescent probe (ER-Tracker Red) staining was used to observe the expression level of reactive oxygen species (ROS) in ER of cells. Mitochondrial membrane potential detection (JC-1) was used to observe cell membrane potential change and early apoptosis. Cell apoptosis was detected by Hoechst33258 staining. The expressions of Nrf2, HO-1 were documented by cell Immunofluorescence staining and Western blot analysis. Results: NBP effectively improved the survival ability of Müller cells shown by MTT assay. NBP effectively alleviated the morphological and apoptotic changes induced by H2O2 stimulation by Calcein AM assay, HE staining, Hoechst 33258, JC-1 staining. H2O2 induction increased the expression level of ROS, whereas, the treatment with NBP could remarkably lower the expression level of ROS. Cell immunofluorescence staining indicated that the fluorescence staining intensity of HO-1 in the NBP group was significantly higher than that in the control group. While the western blotting results showed that the expression level of HO-1 could be increased by NBP in a time-dependent manner. The translocation of Nrf2 in nuclei was observed within 2 h and Nrf2 was identified in nuclei for up to 48 h. Conclusions: Our study demonstrated that NBP had a protective effect on H2O2-induced cytotoxicity in retinal Müller cells in vitro and that it was a potent activator of Nrf2 and HO-1signaling.


Subject(s)
Benzofurans/pharmacology , Ependymoglial Cells/drug effects , Neuroprotective Agents/pharmacology , Oxidative Stress , Retinal Diseases/prevention & control , Apoptosis/drug effects , Blotting, Western , Cell Line , Cell Survival/drug effects , Endoplasmic Reticulum/metabolism , Ependymoglial Cells/metabolism , Fluoresceins/metabolism , Fluorescent Antibody Technique , Heme Oxygenase-1/metabolism , Humans , Hydrogen Peroxide/toxicity , Membrane Potential, Mitochondrial/physiology , Microscopy, Fluorescence , NF-E2-Related Factor 2/metabolism , Oxidants , Reactive Oxygen Species/metabolism , Signal Transduction
3.
Biomed Res Int ; 2019: 1907906, 2019.
Article in English | MEDLINE | ID: mdl-30809532

ABSTRACT

Postmenopausal osteoporosis (PMOP), as well as its associated increased risk for fragility fracture, is one of the most disabling consequences of aging in women. This present study aimed to identify candidate genes that involve pathogenesis of PMOP and the therapeutic mechanism of Liuweidihuang (LWDH) pills on PMOP. We integrated microarray datasets of PMOP derived from the Gene Expression Omnibus (GEO) to screen differentially expressed genes (DEGs) between PMOP and normal controls as well as patients with PMOP and patients after treatment of LWDH pills. GO and KEGG enrichment analysis for DEGs were performed. The shared DEGs, associated with both the pathogenesis of PMOP and the therapeutic mechanism of LWDH, were further analyzed by protein-protein interaction (PPI) network. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to verify the DEGs obtained by our integrated analysis. Compared with normal controls, 1732 DEGs in PMOP were obtained with p<0.05. According to the qRT-PCR results, expression of ATF2, FBXW7, RDX, and RBBP4 was consistent with that in our integrated analysis, generally. GO and KEGG enrichment analysis showed that those DEGs were significantly enriched in regulation of transcription, DNA-dependent, cytoplasm, protein binding, and MAPK signaling pathway. A total of 58 shared DEGs in PMOP versus normal control and in patients with PMOP versus patients after LWDH treatment were identified, which had opposite expression trend in these two comparisons. In the PPI network, CSNK2A1, ATF2, and FBXW7 were three hub proteins. Three genes including ATF2, FBXW7, and RDX were speculated to be therapeutic targets of LWDH for PMOP based on BATMAN-TCM database. We speculated that three genes of ATF2, FBXW7, and RDX may play crucial roles in both pathogenesis of PMOP and therapeutic mechanism of LWDH on PMOP. Our results may provide clues for the molecular pathogenesis of PMOP and offer new possibilities for treatment of PMOP.


Subject(s)
Computational Biology , Drugs, Chinese Herbal/administration & dosage , Osteoporosis, Postmenopausal/drug therapy , Activating Transcription Factor 2/genetics , Cytoskeletal Proteins/genetics , Drugs, Chinese Herbal/adverse effects , F-Box-WD Repeat-Containing Protein 7/genetics , Gene Expression Profiling , Gene Expression Regulation/drug effects , Gene Regulatory Networks/drug effects , Humans , Membrane Proteins/genetics , Osteoporosis, Postmenopausal/genetics , Osteoporosis, Postmenopausal/pathology , Protein Binding/drug effects , Protein Interaction Maps/drug effects , Retinoblastoma-Binding Protein 4/genetics , Signal Transduction/drug effects
6.
Environ Sci Pollut Res Int ; 22(18): 14189-99, 2015 Sep.
Article in English | MEDLINE | ID: mdl-25966886

ABSTRACT

In order to understand the sources and potential formation processes of atmospheric carbonaceous aerosols in South China, fine particle samples were collected at a high-elevation mountain site--Mount Lu (29°35' N, 115°59' E, 1165 m A.S.L.) during August-September, 2011. Eight carbonaceous fractions from particles were resolved following the IMPROVE thermal/optical reflectance protocol. During the observation campaign, the daily concentrations of PM2.5 at Mount Lu ranged from 7.69 to 116.39 µg/m(3), with an average of 58.76 µg/m(3). The observed average organic carbon (OC) and elemental carbon (EC) concentrations in PM2.5 were 3.78 and 1.28 µg/m(3), respectively. Secondary organic carbon (SOC) concentration, estimated by EC-tracer method, was 2.07 µg/m(3) on average, accounting for 45.0% of the total OC. The enhancement of secondary organic aerosol (SOA) formation was observed during cloud/fog processing, and heterogeneous acid-catalyzed reactions may have contributed to SOA formation as well. Back trajectory analysis indicated that air masses were mainly sourced from southern China during observation period, and this air mass source was featured by highest values of OC and effective carbon ratio (ECR). Relation of carbonaceous species and principal component analysis indicated that multiple sources contributed to the carbonaceous aerosols at Mount Lu.


Subject(s)
Air Pollutants/analysis , Air Pollutants/chemistry , Carbon/analysis , Carbon/chemistry , Environmental Monitoring , Organic Chemicals/analysis , Organic Chemicals/chemistry , Aerosols , Atmosphere , China , Motion , Particle Size , Particulate Matter/analysis , Particulate Matter/chemistry
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