scholarly journals WNT11-Conditioned Medium Promotes Angiogenesis through the Activation of Non-Canonical WNT-PKC-JNK Signaling Pathway

Genes ◽  
2020 ◽  
Vol 11 (11) ◽  
pp. 1277
Author(s):  
Jingcai Wang ◽  
Min Gong ◽  
Shi Zuo ◽  
Jie Xu ◽  
Chris Paul ◽  
...  
Keyword(s):  
Cardiac Function ◽  
Conditioned Medium ◽  
Umbilical Vein ◽  
Neutralizing Antibody ◽  
Jnk Signaling ◽  
Paracrine Effects ◽  
Calphostin C ◽  
Jnk Signaling Pathway ◽  
Umbilical Vein Endothelial Cells ◽  
Canonical Wnt

Background: We demonstrated that the transduction of Wnt11 into mesenchymal stem cells (MSCs) (MSCWnt11) promotes these cells differentiation into cardiac phenotypes. In the present study, we investigated the paracrine effects of MSCWnt11 on cardiac function and angiogenesis. Methods and Results: Conditioned medium was collected from MSCWnt11 (CdMWnt11) and their control cells (CdMGFP). CdMWnt11, especially obtained from MSCWnt11 exposed to hypoxia, significantly promoted human umbilical vein endothelial cells (HUVECs) migration and increased capillary-like tube (CLT) formation, which was blocked by Wnt11 neutralizing antibody. Wnt11 protein was significantly higher in CdMWnt11 compared to that in CdMGFP. Directly treating HUVECs with recombinant Wnt11 protein significantly increased CLT formation, which was abrogated by treating cells with the JNK inhibitor SP600125, as well as the PKC inhibitor Calphostin-C. Moreover, the transfection of Wnt11 to HUVECs (HWnt11) significantly increased CLT formation and HUVEC migration, as well as upregulated p-pan-PKC and p-JNK expression. Injection of CdMWnt11 into the peri-infarct region in a rat acute myocardial infarction (AMI) model significantly improved cardiac function, reduced infarct size, and increased myocardial blood flow and blood vessel density in the ischemic area. Conclusion: Wnt11 released from MSCWnt11 increased angiogenesis and improved cardiac function via non-canonical Wnt-PKC-JNK dependent pathways.

Abstract 15272: Wnt11 Delivered in Mesenchymal Stem Cell-Derived Media Promotes Angiogenesis Through Non-canonical Wnt-PKC-JNK Pathway

Circulation ◽  
2014 ◽  
Vol 130 (suppl_2) ◽  
Author(s):  
Jingcai Wang ◽  
Bin Yu ◽  
Min Gong ◽  
Honghai Liu ◽  
Yigang Wang ◽  
...  
Keyword(s):  
Blood Vessel ◽  
Cardiac Function ◽  
Regional Blood Flow ◽  
Umbilical Vein ◽  
Neutralizing Antibody ◽  
Ischemic Myocardium ◽  
Pcr Analysis ◽  
Jnk Pathway ◽  
Umbilical Vein Endothelial Cells

Background: Wnt11 is involved in many developmental and cellular processes. We have previously reported that Wnt11 overexpressing (MSC Wnt11 ) significantly improved cardiac function and increased angiogenesis. Here, we aimed to elucidate how Wnt11 delivered by MSC increase blood vessel formation. Methods and Results: An acute myocardial infarction (MI) model in SD rats was developed by ligation of the left anterior descending (LAD) coronary artery. Conditioned medium (CdM) collected from MSC Wnt11 (CdM Wnt11 ) was injected immediately into the ischemic border area. Rats treated with CdM Wnt11 showed a significantly improved cardiac function. Furthermore, CdM Wnt11 increased blood vessel density and regional blood flow in ischemic myocardium. Fluorescent immunostaining showed that both vWF-positive single endothelial cell numbers and microvessel numbers were significantly increased in CdM Wnt11 treated myocardium. In vitro studies showed that CdM Wnt11 significantly increased number and length of capillary-like structure (CLS) formation and promoted human umbilical vein endothelial cells (HUVECs) migration. These results were confirmed by directly treating HUVECs with recombinant Wnt11 proteins (5μg/ml) or over-expressing Wnt11 in HUVECs (H Wnt11 ). However, these effects could be abolished by using a Wnt11 neutralizing antibody. Immunostaining and Western blotting results revealed upregulation of p-pan-PKC and p-JNK in H Wnt11 . Real-time PCR analysis indicated that the expression of novel PKCs including PKCδ, PKCε, PKCη, and PKCθ was significantly upregulated in H Wnt11 . Moreover, the effect of Wnt11 on CLT formation and HUVECs migration were abrogated when the JNK inhibitor SP600125 at 5 μM and the PKC inhibitor Calphostin-C at 0.1 μM were added to the culture system. Conclusion: Our data suggests, for the first time, that Wnt11 in MSC-derived media improved cardiac function and promoted angiogenesis in ischemic myocardium. The effect of Wnt11 in angiogenesis is mediated by novel PKCs, with JNK as its downstream mediator.

Abstract 346: Involvement of Non-canonical Wnt-PKC-JNK Pathway in Angiogenesis Enhanced by MSC Overexpressing Wnt11

2014 ◽  
Vol 115 (suppl_1) ◽  
Author(s):  
Meifeng Xu ◽  
Jingcai Wang ◽  
Bin Yu
Keyword(s):  
Cardiac Function ◽  
Heart Development ◽  
Umbilical Vein ◽  
Neutralizing Antibody ◽  
Ischemic Myocardium ◽  
Pcr Analysis ◽  
Jnk Pathway ◽  
Protein Levels ◽  
Canonical Wnt

Wnt11 is a key regulator of cardiac muscle cell proliferation and differentiation during heart development. Here, we hypothesized that preprogramming mesenchymal stem cells (MSC) with Wnt11 enhanced angiogenesis in ischemic myocardium. Methods and Results: An acute myocardial infarction model in SD rats was developed by ligation of the left anterior descending (LAD) coronary artery. Rat bone marrow derived MSC were transduced with Wnt11 gene (MSC-Wnt11) and transplanted into ischemic border area. The animals treated with MSC-Wnt11 showed a significantly improved cardiac function. Furthermore, fluorescent microsphere and histological studies revealed an increased blood flow and blood vessel density in MSC-Wnt11 transplanted animals. In vitro studies using conditioned medium (CdM) obtained from MSC-Wnt11 (CdM-Wnt11) showed an increased the length and number of capillary-like structure (CLS) formation and promoted migration of human umbilical vein endothelial cells (HUVECs), which was similar to that HUVECs were directly treated with recombinant Wnt11 proteins. However, these effects could be abolished by using a Wnt11 neutralizing antibody. Real-time PCR analysis indicated that the expression of novel PKCs including PKCδ, PKCε, PKCη, and PKCθ was significantly upregulated in HUVECs transduced with Wnt11 (H-Wnt11). Immunostaining and western blotting results showed that the protein levels of p-pan-PKC and p-JNK as well as VEGFA in H-Wnt11 were significantly higher than that in control HUVECs. However, no difference was detected in the expression of either p-p38 or p-ERK between H-Wnt11 and its control. Furthermore, the enhanced CLS formation and migration of HUVECs mediated by CdM-Wnt11 were partially abolished by using JNK inhibitor, SP600125 (5 μM) and PKC inhibitor, Calphostin-C (0.1 μM). Conclusion: Our results demonstrated, for the first time, that Wnt11 delivered by MSC improved cardiac function and promotes angiogenesis in ischemic myocardium, which may be associated with the activation of the non-canonical Wnt-PKC-JNK pathway.

scholarly journals Preeclampsia Status Controls Interleukin-6 and Soluble IL-6 Receptor Release from Neutrophils and Endothelial Cells: Relevance to Increased Inflammatory Responses

2021 ◽  
Vol 28 (2) ◽  
pp. 202-211
Author(s):  
Yuping Wang ◽  
Yang Gu ◽  
J. Steven Alexander ◽  
David F. Lewis
Keyword(s):  
Endothelial Cells ◽  
Conditioned Medium ◽  
Interleukin 6 ◽  
Inflammatory Responses ◽  
Vascular System ◽  
Umbilical Vein ◽  
Normal Pregnancy ◽  
Hypertensive Disorder ◽  
Umbilical Vein Endothelial Cells

Increased neutrophil–endothelial binding and inflammatory responses are significant pathophysiological events in the maternal vascular system in preeclampsia, a hypertensive disorder in human pregnancy. Interleukin 6 (IL-6) and its soluble receptors (soluble IL-6R (sIL-6R) and soluble gp130 (sgp130)) are critical inflammatory mediators. During pregnancy, maternal IL-6 and sgp130 levels were increased, but sIL-6R levels were decreased, in women with preeclampsia compared to normotensive pregnant women. However, little is known about differences in IL-6, sIL-6R, and sgp130 production by neutrophils and endothelial cells between normal pregnancy and preeclampsia. To study this, we isolated neutrophils and cultured human umbilical vein endothelial cells (HUVECs) from normal and preeclamptic pregnancies. Production of IL-6, sIL-6R, and sgp130 was measured. The role of placental factor(s)-mediated neutrophil production of IL-6, sIL-6R, and sgp130 was also determined by pretreating neutrophils with placental conditioned medium generated from placental villous cultures. We found that IL-6 and sgp130 were mainly produced by endothelial cells, while sIL-6R was mainly produced by neutrophils. Endothelial cells from preeclampsia produced significantly more IL-6 and sgp130, and neutrophils from preeclampsia produced significantly less sIL-6R than normal pregnancy cells. Interestingly, production of IL-6, sIL-6R, and sgp130 were time-dependently increased when neutrophils and endothelial cells were co-cultured. We also found that neutrophils from normal pregnancies produced more IL-6, but less sIL-6R, after being primed by preeclamptic-placental conditioned medium. These results demonstrated that neutrophils and endothelial cells have different capacities in producing IL-6, sIL-6R, and sgp130 between normal pregnancy and preeclampsia. These results also provide evidence that the placenta plays a role in inducing neutrophil activation in preeclampsia.

scholarly journals Prostate-Specific Membrane Antigen (PSMA) Promotes Angiogenesis of Glioblastoma Through Interacting With ITGB4 and Regulating NF-κB Signaling Pathway

2021 ◽  
Vol 9 ◽  
Author(s):  
Yang Gao ◽  
Hui Zheng ◽  
Liangdong Li ◽  
Mingtao Feng ◽  
Xin Chen ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Conditioned Medium ◽  
Poor Prognosis ◽  
Umbilical Vein ◽  
Tube Formation ◽  
Prostate Specific Membrane Antigen ◽  
Umbilical Vein Endothelial Cells ◽  
Specific Membrane

BackgroundGlioblastoma multiforme (GBM) is the most common primary malignant tumor in the central nervous system (CNS), causing the extremely poor prognosis. Combining the role of angiogenesis in tumor progression and the role of prostate-specific membrane antigen (PSMA) in angiogenesis, this study aims to explore the functions of PSMA in GBM.MethodsClinical GBM specimens were collected from 60 patients who accepted surgical treatment in Fudan University Shanghai Cancer Center between January 2018 and June 2019. Immunohistochemical staining was used to detect PSMA and CD31 expression in GBM tissues. Prognostic significance of PSMA was evaluated by bioinformatics. Human umbilical vein endothelial cells (HUVECs) transfected with PSMA overexpression plasmids or cultured with conditioned medium collected based on GBM cells, were used for CCK8, Transwell and tube formation assays. High-throughput sequencing and immunoprecipitation were used to explore the underlying mechanism. Furthermore, the in vivo experiment had been also conducted.ResultsWe demonstrated that PSMA was abundantly expressed in endothelium of vessels of GBM tissues but not in vessels of normal tissues, which was significantly correlated with poor prognosis. Overexpression of PSMA could promotes proliferation, invasion and tube formation ability of human umbilical vein endothelial cells (HUVECs). Moreover, U87 or U251 conditioned medium could upregulated PSMA expression and induce similar effects on phenotypes of HUVECs, all of which could be partially attenuated by 2-PMPA treatment. The mechanistic study revealed that PSMA might promote angiogenesis of GBM through interacting with Integrin β4 (ITGB4) and activating NF-κB signaling pathway. The in vivo growth of GBM could be alleviated by the treatment of 2-PMPA.ConclusionThis study identified PSMA as a critical regulator in angiogenesis and progression of GBM, which might be a promising therapeutic target for GBM treatment.

scholarly journals Advanced Glycation End Products Inhibit Production and Activity of Matrix Metalloproteinase-2 in Human Umbilical Vein Endothelial Cells

2007 ◽  
Vol 35 (5) ◽  
pp. 709-715 ◽  
Author(s):  
L Gao ◽  
L Kang ◽  
Q Chen ◽  
C Chen ◽  
B Xu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Endothelial Cells ◽  
Conditioned Medium ◽  
Advanced Glycation End Products ◽  
Umbilical Vein ◽  
Advanced Glycation ◽  
Glycation End Products ◽  
End Products ◽  
Umbilical Vein Endothelial Cells ◽  
Expression And Activity

The aim of this study was to investigate the effects of advanced glycation end products (AGEs) on the expression and activity of matrix metalloproteinases-2 (MMP-2) in human umbilical vein endothelial cells (HUVECs). Cultured HUVECs were incubated with various concentrations of AGEs-modified albumin or unmodified albumin for different time periods. Protein and gene expression of MMP-2 and the receptor for AGEs (RAGE) were measured by Western blot and reverse transcription-polymerase chain reaction, respectively. The activity of MMP-2 in the conditioned medium was measured by gelatin zymography. The AGE-modified albumin inhibited MMP-2 but increased RAGE protein and gene expression in HUVECs in a concentration- and time-dependent manner. An inhibition of MMP-2 activity was also detected in the conditioned medium of HUVECs incubated with AGEs-modified albumin. In conclusion, AGEs inhibited the expression and activity of MMP-2 in HUVECs; this may be mediated through upregulation of RAGE.

scholarly journals The Stemness-High Human Colorectal Cancer Cells Promote Angiogenesis by Producing Higher Amounts of Angiogenic Cytokines via Activation of the Egfr/Akt/Nf-κB Pathway

2021 ◽  
Vol 22 (3) ◽  
pp. 1355
Author(s):  
Shin-Yi Chung ◽  
Ta-Chung Chao ◽  
Yeu Su
Keyword(s):  
Colorectal Cancer ◽  
Cancer Metastasis ◽  
Umbilical Vein ◽  
Neutralizing Antibody ◽  
Tube Formation ◽  
Conditioned Media ◽  
Vascular Density ◽  
Human Colorectal Cancer ◽  
Colorectal Cancer Cells ◽  
Umbilical Vein Endothelial Cells

Purpose: Cancer stem cells (CSCs) are responsible for cancer metastasis by stimulating tumor angiogenesis via various mechanisms. To elucidate the potential of the stemness-high human colorectal cancer (CRC) cells (i.e., CRCSCs) in activating angiogenesis, effects of the GATA6-overexpressing HCT-116 and HT-29 human CRC clones established previously by us in promoting the angiogenesis of human umbilical vein endothelial cells (HUVECs) were examined. Methods: Angiogenesis-promoting effects (i.e., migration, invasion, DNA synthesis, and tube formation) in HUVECs of the conditioned media (CM) from various human CRC clones were analyzed. MMP activities were assessed using a zymography assay. Western blotting and selective inhibitors were used to dissect the signaling pathway involved. IHC was used to examine the vascular density in tumor xenografts. Results: We found that the conditioned media (CM) collected from the GATA6-overexpressing clones enhanced angiogenesis of HUVECs more effectively which might be attributed partly to a higher MMP-9 production by HUVECs. Subsequently, elevated levels of IL-8 and VEGF-A were detected in the CM whose tube formation-enhancing activities were abolished by the co-treatment with either a VEGFR2 inhibitor or an IL-8 neutralizing antibody. Interestingly, increased production of these cytokines in the GATA6-overexpressing clones was due to an EGFR/AKT-mediated activation of NF-κB. Furthermore, not only were the levels of CD31 and endomucin but also the blood vessel density was much higher in the xenograft tumors grown from these clones. Conclusion: Our findings demonstrate that human CRCSCs promote a stronger angiogenesis by producing higher amounts of angiogenic factors through activation of the EGFR/AKT/NF-κB pathway.

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