Anti-inflammatory Role of Carotenoids in Endothelial Cells Derived from Umbilical Cord of Women Affected by Gestational Diabetes Mellitus

Diabetes is associated with vascular inflammation, endothelial dysfunction, and oxidative stress, promoting the development of cardiovascular diseases (CVD). Several studies showed that a carotenoid-rich diet is associated to a reduced cardiovascular risk in healthy and diabetic subjects, although the mechanisms of action are still unknown. Here, the potential role of β-carotene (BC) and lycopene (Lyc) in human endothelial cells isolated from human umbilical cord vein (HUVECs) of women with gestational diabetes (GD) and respective controls (C) has been investigated. Results showed that BC and Lyc reduced the tumor necrosis factor alpha- (TNF-α-) stimulated monocyte-endothelium interaction (adhesion assay), membrane exposure (flow cytometry), and total expression levels (Western blot) of VCAM-1 and ICAM-1 in both cell types. Moreover, the treatment with BC and Lyc reduced the TNF-α-induced nuclear translocation of NF-κB (image flow cytometry) by preserving bioavailability of nitric oxide (NO, flow cytometry, and cGMP EIA kit assay), a key vasoactive molecule. Notably, BC and Lyc pretreatment significantly reduced peroxynitrite levels (flow cytometry), contributing to the redox balance protection. These results suggest a new mechanism of action of carotenoids which exert vascular protective action in diabetic condition, thus reinforcing the importance of a carotenoid-rich diet in the prevention of diabetes cardiovascular complications.

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Overexpression of Pygo2 Increases Differentiation of Human Umbilical Cord Mesenchymal Stem Cells into Cardiomyocyte-like Cells

Current Molecular Medicine ◽

10.2174/1566524019666191017150416 ◽

2020 ◽

Vol 20 (4) ◽

pp. 318-324 ◽

Cited By ~ 3

Author(s):

Lei Yang ◽

Shuoji Zhu ◽

Yongqing Li ◽

Jian Zhuang ◽

Jimei Chen ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Umbilical Cord ◽

Heart Function ◽

Critical Role ◽

Human Umbilical Cord ◽

Stem Cell Markers ◽

Quantitative Real Time Pcr ◽

Qrt Pcr

Background: Our previous studies have shown that Pygo (Pygopus) in Drosophila plays a critical role in adult heart function that is likely conserved in mammals. However, its role in the differentiation of human umbilical cord mesenchymal stem cells (hUC-MSCs) into cardiomyocytes remains unknown. Objective: To investigate the role of pygo2 in the differentiation of hUC-MSCs into cardiomyocytes. Methods: Third passage hUC-MSCs were divided into two groups: a p+ group infected with the GV492-pygo2 virus and a p− group infected with the GV492 virus. After infection and 3 or 21 days of incubation, Quantitative real-time PCR (qRT-PCR) was performed to detect pluripotency markers, including OCT-4 and SOX2. Nkx2.5, Gata-4 and cTnT were detected by immunofluorescence at 7, 14 and 21 days post-infection, respectively. Expression of cardiac-related genes—including Nkx2.5, Gata-4, TNNT2, MEF2c, ISL-1, FOXH1, KDR, αMHC and α-Actin—were analyzed by qRT-PCR following transfection with the virus at one, two and three weeks. Results : After three days of incubation, there were no significant changes in the expression of the pluripotency stem cell markers OCT-4 and SOX2 in the p+ group hUC-MSCs relative to controls (OCT-4: 1.03 ± 0.096 VS 1, P > 0.05, SOX2: 1.071 ± 0.189 VS 1, P > 0.05); however, after 21 days, significant decreases were observed (OCT-4: 0.164 ± 0.098 VS 1, P < 0.01, SOX2: 0.209 ± 0.109 VS 1, P < 0.001). Seven days following incubation, expression of mesoderm specialisation markers, such as Nkx2.5, Gata-4, MEF2c and KDR, were increased; at 14 days following incubation, expression of cardiac genes, such as Nkx2.5, Gata-4, TNNT2, MEF2c, ISL-1, FOXH1, KDR, αMHC and α-Actin, were significantly upregulated in the p+ group relative to the p− group (P < 0.05). Taken together, these findings suggest that overexpression of pygo2 results in more hUCMSCs gradually differentiating into cardiomyocyte-like cells. Conclusion: We are the first to show that overexpression of pygo2 significantly enhances the expression of cardiac-genic genes, including Nkx2.5 and Gata-4, and promotes the differentiation of hUC-MSCs into cardiomyocyte-like cells.

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Exosomes from human umbilical cord mesenchymal stem cells attenuate the inflammation of severe steroid-resistant asthma by reshaping macrophage polarization

Stem Cell Research & Therapy ◽

10.1186/s13287-021-02244-6 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Bing Dong ◽

Chao Wang ◽

Jing Zhang ◽

Jinrong Zhang ◽

Yinuo Gu ◽

...

Keyword(s):

Flow Cytometry ◽

Stem Cell ◽

Mesenchymal Stem Cell ◽

Quantitative Proteomics ◽

Macrophage Polarization ◽

Human Umbilical Cord ◽

Raw 264.7 ◽

Steroid Resistant ◽

Central Protein

Abstract Background Severe, steroid-resistant asthma (SSRA) is a serious clinical problem in asthma management. Affected patients have severe clinical symptoms, worsened quality of life, and do not respond to steroid, a mainstay steroid treatment of asthma. Thus, effective therapies are urgently needed. Exosomes derived from mesenchymal stem cell (MSC-Exo) has become attractive candidates for the lung inflammatory diseases through its immunomodulatory effects. In this study, we explored the therapeutic effects of MSC-Exo in SSRA and identified the therapeutic mechanism of MSC-Exo. Method Exosomes from human umbilical cord mesenchymal stem cell (hUCMSC) were isolated and characterized by transmission electron microscopy, nanoparticle tracking analysis and flow cytometry analysis. Effects of MSC-Exo on airway hyper responsiveness (AHR), inflammation, histopathology, and macrophage polarization in SSRA in mice were evaluated. Systematic depletion of macrophages determined the role of macrophages in the therapeutic effect of SSRA in mice. LPS-stimulated RAW 264.7 cell model was constructed to determine the underlying mechanism of MSC-Exo on macrophage polarization. qRT-PCR, Western blotting, immunofluorescence, and flow cytometry were performed to evaluate the expression of M1 or M2 markers. Tandem mass tags (TMT)-labeled quantitative proteomics were applied to explore the central protein during the regulation effect of MSC-Exo on macrophage polarization. Knockdown and overexpression of TRAF1 were used to further clarify the role of the central protein on macrophage polarization. Result We successfully isolated and characterized exosomes from hUCMSCs. We verified that the intratracheal administration of MSC-Exo reversed AHR, histopathology changes, and inflammation in SSRA mice. Systematic depletion of macrophages weakened the therapeutic effect of MSC-Exo. We found that MSC-Exo treatment inhibited M1 polarization and promoted M2 polarization in LPS-stimulated RAW 264.7 cells. Subsequently, tumor necrosis factor receptor-associated factor 1 (TRAF1) was determined as the central protein which may be closely related to the regulation of macrophage polarization from TMT-labeled quantitative proteomics analysis. Knockdown and overexpression of TRAF1 demonstrated that the effect of MSC-Exo treatment on macrophage polarization, NF-κB and PI3K/AKT signaling was dependent on TRAF1. Conclusion MSC-Exo can ameliorate SSRA by moderating inflammation, which is achieved by reshaping macrophage polarization via inhibition of TRAF1.

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Venous and Arterial Endothelial Cells from Human Umbilical Cords: Potential Cell Sources for Cardiovascular Research

International Journal of Molecular Sciences ◽

10.3390/ijms22020978 ◽

2021 ◽

Vol 22 (2) ◽

pp. 978

Author(s):

Skadi Lau ◽

Manfred Gossen ◽

Andreas Lendlein ◽

Friedrich Jung

Keyword(s):

Endothelial Cells ◽

Umbilical Cord ◽

Membrane Integrity ◽

Cell Types ◽

Cell Number ◽

Human Umbilical Cord ◽

Cardiovascular Research ◽

Cardiovascular Devices ◽

Arterial Endothelial Cells

Although cardiovascular devices are mostly implanted in arteries or to replace arteries, in vitro studies on implant endothelialization are commonly performed with human umbilical cord-derived venous endothelial cells (HUVEC). In light of considerable differences, both morphologically and functionally, between arterial and venous endothelial cells, we here compare HUVEC and human umbilical cord-derived arterial endothelial cells (HUAEC) regarding their equivalence as an endothelial cell in vitro model for cardiovascular research. No differences were found in either for the tested parameters. The metabolic activity and lactate dehydrogenase, an indicator for the membrane integrity, slightly decreased over seven days of cultivation upon normalization to the cell number. The amount of secreted nitrite and nitrate, as well as prostacyclin per cell, also decreased slightly over time. Thromboxane B2 was secreted in constant amounts per cell at all time points. The Von Willebrand factor remained mainly intracellularly up to seven days of cultivation. In contrast, collagen and laminin were secreted into the extracellular space with increasing cell density. Based on these results one might argue that both cell types are equally suited for cardiovascular research. However, future studies should investigate further cell functionalities, and whether arterial endothelial cells from implantation-relevant areas, such as coronary arteries in the heart, are superior to umbilical cord-derived endothelial cells.

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Simultaneous isolation of vascular endothelial cells and mesenchymal stem cells from the human umbilical cord

In Vitro Cellular & Developmental Biology - Animal ◽

10.1007/s11626-008-9155-4 ◽

2008 ◽

Vol 45 (1-2) ◽

pp. 23-27 ◽

Cited By ~ 29

Author(s):

Sachin S. Kadam ◽

Shubha Tiwari ◽

Ramesh R. Bhonde

Keyword(s):

Stem Cells ◽

Endothelial Cells ◽

Mesenchymal Stem Cells ◽

Umbilical Cord ◽

Vascular Endothelial Cells ◽

Human Umbilical Cord ◽

Vascular Endothelial

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Lysophospholipids increase IL-8 and MCP-1 expressions in human umbilical cord vein endothelial cells through an IL-1-dependent mechanism

Journal of Cellular Biochemistry ◽

10.1002/jcb.20963 ◽

2006 ◽

Vol 99 (4) ◽

pp. 1216-1232 ◽

Cited By ~ 63

Author(s):

Chi Iou Lin ◽

Chiung-Nien Chen ◽

Jiun Hong Chen ◽

Hsinyu Lee

Keyword(s):

Endothelial Cells ◽

Umbilical Cord ◽

Human Umbilical Cord ◽

Umbilical Cord Vein ◽

Human Umbilical Cord Vein ◽

Dependent Mechanism

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Demonstration of Antithrombin III in Fresh and Cultured Endothelial Cells from Human Umbilical Cord

10.1055/s-0039-1684362 ◽

1979 ◽

Author(s):

Vivian Chan ◽

T.K. Chan

Keyword(s):

Endothelial Cells ◽

Umbilical Cord ◽

Antithrombin Iii ◽

Cultured Cells ◽

Active Role ◽

Human Umbilical Cord ◽

Thrombosis Research ◽

Natural Inhibitor ◽

Immunofluorescent Technique ◽

Radio Immunoassay

We have shown by immunofluorescent technique that the distribution of antithrombin III (ATIII) in human tissues was concentrated around the microvasculature of the lungs and kidneys, as well as veins and small arteries of other organs (liver and spleen). It would seem that ATIII is stored and/or synthesized in the endothelial cells similar to Factor VIII-RAG and Plasminogen Activator. Endothelial cells were isolated from human umbilical cord by collagenase and cultured according to Chemethod described by Shearn etal (1977). In freshly isolated endothelial cells, ATIII could be demonstrated by indirect immunof1uorescent technique and radio immunoassay confirmed the presence of 14.8 ng per 106 cells. After 7 days’ culture, the supernatant from 106 cells contained about 15 ng and the cultured cells (106) contained 16.9 ng ATIII. The presence of ATIII in cultured cells was also confirmed by the positive immunofluorescence. Hence the endothelial cells play an active role in preventing thrombosis by the synthesis and liberation of ATIII, the major natural inhibitor of the intrinsic pathway of Coagulation.Reference: Shearn S.A., Peake I.R., Ciddings J.C., Humphrys J. and Bloom A.L. Thrombosis Research, 11, 43, 1977.

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Effect Of Thrombin On Platelet Adherence To Cultured Vascular Cells: Modification By Dapa

10.1055/s-0038-1652167 ◽

1981 ◽

Author(s):

R L Czervionke ◽

J C Hoak ◽

D L Haycraft ◽

G L Fry

Keyword(s):

Endothelial Cells ◽

Platelet Activation ◽

Human Umbilical Cord ◽

Vascular Cells ◽

Aggregate Formation ◽

Platelet Adherence ◽

Vascular Origin ◽

Fast Acting ◽

Positive Controls

The role of thrombin in inducing adherence of platelets to cultured vascular cells is complex and may involve factors of both platelet and vascular origin. Dansylarginine N- (3-ethyl-l,5-pentanediyl)amide, DAPA, a fast-acting inhibitor of thrombin, was used in this study to evaluate platelet activation and endothelial PGI2 release induced by thrombin. Monolayers of endothelial cells and fibroblasts were cultured from human umbilical cord vessels. Empty culture dishes were used as a control. Some vascular cells were treated with aspirin (ASA) to inhibit PGI2 formation before they were incubated with 51Cr-platelets for platelet adherence studies. PGI2 was assayed by radioimmunoassay for 6-keto-PGF1α. In one study, endothelium was incubated with 0.3 U thrombin for 2 min. before 10 μM DAPA was added. Platelets were next mixed with this solution and adherence was determined. DAPA caused a decrease in thrombin-induced platelet adherence to normal endothelium from 8% to 2%. When ASA- treated endothelium was employed, DAPA decreased adherence from 59% to 2%. In another study, 51cr-platelets were first aggregated with 0.1 U thrombin, then 1.3 μM DAPA was added, and these aggregates were incubated with the monolayers. Similar experiments without DAPA served as positive controls. DAPA did not reverse the aggregate formation, but it did reduce platelet adherence to normal endothelium from 38% to 1%, and to ASA-treated endothelium from 68% to 11%. In contrast, DAPA had little effect with fibroblasts (81% to 71%), ASA-treated fibroblasts (82% to 67%), and the empty dish control (86% to 66%). These results suggest that thrombin-induced platelet adherence in this system involves more than just an effect upon platelets. In addition, they provide further evidence that the non-thrombogenic nature of endothelium persists despite the absence of PGI2.

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Human Umbilical Cord Mesenchymal Stem Cells Inhibit the Function of Allogeneic Activated Vγ9Vδ2 T Lymphocytes In Vitro

BioMed Research International ◽

10.1155/2015/317801 ◽

2015 ◽

Vol 2015 ◽

pp. 1-10 ◽

Cited By ~ 4

Author(s):

Xiaohuan Liu ◽

Ting Feng ◽

Tianxiang Gong ◽

Chongyang Shen ◽

Tingting Zhu ◽

...

Keyword(s):

Stem Cells ◽

Flow Cytometry ◽

T Cells ◽

Mesenchymal Stem Cells ◽

T Cell ◽

Umbilical Cord ◽

Interleukin 2 ◽

Cell Contact ◽

Human Umbilical Cord ◽

Fas L

Background. Human umbilical cord mesenchymal stem cells (UC-MSCs) can regulate the function of immune cells. However, whether and how UC-MSCs can modulate the function of Vγ9Vδ2 T cells has not been fully understood. Methods. The PBMCs or Vγ9Vδ2 T cells were activated and expanded with pamidronate (PAM) and interleukin-2 (IL-2) with or without the presence UC-MSCs. The effects of UC-MSCs on the proliferation, cytokine expression, and cytotoxicity of Vγ9Vδ2 T cells were determined by flow cytometry. The effects of UC-MSCs on Fas-L, TRAIL-expressing Vγ9Vδ2 T cells, and Vγ9Vδ2 T cell apoptosis were determined by flow cytometry. Results. UC-MSCs inhibited Vγ9Vδ2 T cell proliferation in a dose-dependent but cell-contact independent manner. Coculture with UC-MSCs reduced the frequency of IFNγ+ but increased granzyme B+ Vγ9Vδ2 T cells. UC-MSCs inhibited the cytotoxicity of Vγ9Vδ2 T cells against influenza virus H1N1 infected A549 cells and also reduced the frequency of Fas-L+, TRAIL+ Vγ9Vδ2 T cells but failed to modulate the apoptosis of Vγ9Vδ2 T cells. Conclusions. These results indicated that UC-MSCs efficiently suppressed the proliferation and cytotoxicity of Vγ9Vδ2 T cells and modulated their cytokine production. Fas-L and TRAIL were involved in the regulation. Cell contact and apoptosis of Vγ9Vδ2 T cells were not necessary for the inhibition.

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