Comparative proteomic analysis of membrane microdomains isolated from two hyperlipidemic animal models

Uyy, Elena; Boteanu, Raluca Maria; Ivan, Luminiţa; Antohe, Felicia

doi:10.1016/j.bbapap.2016.05.009

Cited by 7 publications

(5 citation statements)

References 42 publications

(54 reference statements)

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“…The cells were washed twice with PBS, centrifuged and the pellet was solubilized in a denaturant buffer containing 8 M urea, 1% DOC, 0.1% Tris-HCl (pH 8.8) and protease inhibitors, using sonication pulses for 30 s, on ice (UP50H from Hielscher, Teltow, Germany). Protein purification, carbamydomethylation, proteolysis and solid-phase extraction were performed as previously described (Suica et al 2016).…”

Section: Liquid Chromatography-mass Spectrometry (Lc-ms) Analysis Of Mscmentioning

confidence: 99%

Dihydrotestosterone induces pro-angiogenic factors and assists homing of MSC into the cardiac tissue

Popa

Mihai

Constantin

et al. 2018

Journal of Molecular Endocrinology

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The use of mesenchymal stem cells (MSC) as a therapeutic tool in cardiovascular diseases is promising. Since androgens exert some beneficial actions on the cardiovascular system, we tested our hypothesis that this hormone could promote MSC-mediated repair processes, also. Cultured MSCs isolated from Wharton's jelly were exposed to 30 nM dihydrotestosterone (DHT) for 1 or 4 days and the effects of the hormone on their growth/migration/adhesion and the underlying mechanisms were assessed. Results were obtained by real-time cell impedance measurements, and DNA quantification showed that DHT increased MSC proliferation by ~30%. As determined by xCELLigence system, DHT augmented (~2 folds) the migration of MSC toward cardiac tissue slices (at 12 h), and this effect was blocked by flutamide, an androgen receptor (AR) antagonist. Exposure of cells to DHT, upregulated the gene and protein expression of , and and downregulated the expression of DHT significantly induced the release of nitric oxide by MSC (≥2-fold) and flutamide blocked this effect. When MSCs were co-cultured with cardiac slices, immunohistochemical analysis and qRT-PCR showed that the integration of DHT-stimulated MSC was significantly higher than that of in controls. In conclusion, our findings provide the first evidence that DHT promotes MSC growth, migration and integration into the cardiac slices. The modulating effects of DHT were associated with upregulation of ARs and of key molecules known to promote tissue remodeling and angiogenesis. Our findings suggest that priming of MSC with DHT may potentially increase their capability to regenerate cardiac tissue; studies are needed to confirm our findings.

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Section: Liquid Chromatography-mass Spectrometry (Lc-ms) Analysis Of Mscmentioning

confidence: 99%

Dihydrotestosterone induces pro-angiogenic factors and assists homing of MSC into the cardiac tissue

Popa

Mihai

Constantin

et al. 2018

Journal of Molecular Endocrinology

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“…Lipid metabolism of hamsters have been reported that closely resemble to human beings. Thus, hamsters were usually considered as the first and appropriate animal model for estimating hypolipidemic effects ( Suica et al, 2016 ). Male Golden Syrian hamsters (6 weeks old) were purchased from the National Laboratory Animal Center (NLAC), Taipei City, Taiwan.…”

Section: Methodsmentioning

confidence: 99%

Lipid Metabolism and its Mechanism Triggered by Supercritical CO2 Extract of Adlay (Coix lacryma-jobi var. ma-yuen (Rom. Caill.) Stapf) Bran in High-Fat Diet Induced Hyperlipidemic Hamsters

et al. 2021

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Adlay (Coix lacryma-jobi var. ma-yuen (Rom. Caill.) Stapf) seeds are edible crop classified as Traditional Chinese Medicine (TCM). Adlay bran (AB) is one of the wastes generated during adlay refining processes. In this work, supercritical fluid extract of AB (AB-SCF) was investigated to reveal its lipid regulating potential and decode its bifunctional ingredients. AB-SCF×0.5 (30.84 mg/kg/body weight), AB-SCF×1 (61.67 mg/kg/BW), AB-SCF×5 (308.35 mg/kg/BW) and AB-SCF×10 (616.70 mg/kg/BW) were administrated to high fat-diet (HFD) induced hyperglycemic hamsters for 8 weeks. The results indicates that AB-SCF displays a prevention of dramatic body weight gains, lower levels of serum TG, TC, LDL-C and higher in HDL-C, amelioration of cardiovascular risk, alleviation of hepatic TG, TC and lipid peroxidation, and enhancement on cholesterol metabolism with higher bile acid excretion. Investigations on energy metabolic mechanism demonstrates that the hyperlipidemia mitigating capacities of AB-SCF are up-regulated on lipoprotein lipase, AMPK, p-AMPK and down-regulated at fatty acid synthase. Major bio-functional lipid compositions are identified as linoleic acid (28.59%) and oleic acid (56.95%). Non-lipid chemical and active markers are confirmed as 3-O-(trans-4-feruloyl)-β-sitostanol (1463.42 ppm), 3-O-(cis-4-feruloyl)-β-sitostanol (162.60 ppm), and β-sitosterol (4117.72 ppm). These compositions might synergistically responsible for the mentioned activities and can be regarded as analytical targets in quality control. AB-SCF may be considered as a promising complementary supplement, and developed as a functional food or new botanical drug in the future.

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“…We rigorously followed protocols as previously described [ 15 ]. The LC–MS analysis was performed using EASY n-LC II system (Thermo Fisher Scientific, Carlsbad, CA, USA) while mass spectra were obtained with a 60k-LTQ mass spectrometer—Orbitrap Velos Pro (Thermo Fisher Scientific, Carlsbad, CA, USA) [ 24 ].…”

Section: Methodsmentioning

confidence: 99%

Proteomic Analysis of Estrogen-Mediated Enhancement of Mesenchymal Stem Cell-Induced Angiogenesis In Vivo

Mihai

Popa

Suica

et al. 2021

Cells

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Therapeutic use of mesenchymal stem cells (MSCs) for tissue repair has great potential. MSCs from multiple sources, including those derived from human umbilical matrix, namely Wharton’s jelly, may serve as a resource for obtaining MSCs. However, low in vivo engraftment efficacy of MSCs remains a challenging limitation. To improve clinical outcomes using MSCs, an in-depth understanding of the mechanisms and factors involved in successful engraftment is required. We recently demonstrated that 17β-estradiol (E2) improves MSCs in vitro proliferation, directed migration and engraftment in murine heart slices. Here, using a proteomics approach, we investigated the angiogenic potential of MSCs in vivo and the modulatory actions of E2 on mechanisms involved in tissue repair. Specifically, using a Matrigel® plug assay, we evaluated the effects of E2 on MSCs-induced angiogenesis in ovariectomized (OVX) mice. Moreover, using proteomics we investigated the potential pro-repair processes, pathways, and co-mechanisms possibly modified by the treatment of MSCs with E2. Using RT-qPCR, we evaluated mRNA expression of pro-angiogenic molecules, including endoglin, Tie-2, ANG, and VEGF. Hemoglobin levels, a marker for blood vessel formation, were increased in plugs treated with E2 + MSCs, suggesting increased capillary formation. This conclusion was confirmed by the histological analysis of capillary numbers in the Matrigel® plugs treated with E2 + MSC. The LC-MS screening of proteins obtained from the excised Matrigel® plugs revealed 71 proteins that were significantly altered following E2 exposure, 57 up-regulated proteins and 14 down-regulated proteins. A major result was the association of over 100 microRNA molecules (miRNAs) involved in cellular communication, vesicle transport, and metabolic and energy processes, and the high percentage of approximately 25% of genes involved in unknown biological processes. Together, these data provide evidence for increased angiogenesis by MSCs treated with the sex hormone E2. In conclusion, E2 treatment may increase the engraftment and repair potential of MSCs into tissue, and may promote MSC-induced angiogenesis after tissue injury.

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Comparative proteomic analysis of membrane microdomains isolated from two hyperlipidemic animal models

Cited by 7 publications

References 42 publications

Dihydrotestosterone induces pro-angiogenic factors and assists homing of MSC into the cardiac tissue

Dihydrotestosterone induces pro-angiogenic factors and assists homing of MSC into the cardiac tissue

Lipid Metabolism and its Mechanism Triggered by Supercritical CO2 Extract of Adlay (Coix lacryma-jobi var. ma-yuen (Rom. Caill.) Stapf) Bran in High-Fat Diet Induced Hyperlipidemic Hamsters

Proteomic Analysis of Estrogen-Mediated Enhancement of Mesenchymal Stem Cell-Induced Angiogenesis In Vivo

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