Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2021 Oct 27;30(12):1543-1553.
doi: 10.1007/s10068-021-00990-0. eCollection 2021 Nov.

DHA-enriched phosphatidylcholine suppressed angiogenesis by activating PPARγ and modulating the VEGFR2/Ras/ERK pathway in human umbilical vein endothelial cells

Yuanyuan Liu  1 , Yingying Tian  1 , Yao Guo  1 , Ziyi Yan  1 , Changhu Xue  1   2 , Jingfeng Wang  1
Affiliations

DHA-enriched phosphatidylcholine suppressed angiogenesis by activating PPARγ and modulating the VEGFR2/Ras/ERK pathway in human umbilical vein endothelial cells

Yuanyuan Liu et al. Food Sci Biotechnol. .

Abstract

Docosahexaenoic acid-enriched phosphatidylcholine (DHA-PC) is a new generation of omega-3 lipids, which contains an ester bond linking DHA at the sn-2 position of phospholipid. DHA-PC has become the interest recently as its better bioavailability and anti-oxidation capacity. In this study, the anti-angiogenic effect of DHA-PC was evaluated. The capacities of proliferation, migration, tube formation of human umbilical vein endothelial cells were significantly declined after DHA-PC treatment. Furthermore, DHA-PC inhibited the neovascularization of the chick chorioallantoic membrane in vivo. Mechanism results indicated that DHA-PC enhances the expression of peroxisome proliferator-activated receptor γ (PPARγ) at transcriptional and translational level, subsequently down-regulates the VEGFR2 expression and VEGFR2-mediated downstream Ras/ERK pathway, resulting in significant reduction in proliferation and differentiation. Additionally, PPARγ-specific antagonist GW9662 partly reversed the inhibition effects of DHA-PC on tube formation and neovascularization, suggesting that DHA-PC exerts anti-angiogenesis effect through activating PPARγ. These findings indicated that DHA-PC has a great prospect of anti-tumor angiogenesis therapy.

Keywords: Angiogenesis; DHA-PC; Neovascularization; PPARγ; Tube formation.

PubMed Disclaimer

Conflict of interest statement

Conflict of interestNo potential conflicts of interest.

Figures

Fig. 1
Fig. 1
(A) The chemical structure of DHA-PC. DHA esterified at the sn-2 position of glycerol, the medium-chain fatty acids (MCFAs) esterified at the sn-1 position, and the polar group choline boned at the sn-3 position of glycerol. (B) Gas chromatographic analysis of phospholipids isolated from squid roe. (C) The MTT assay. (D) The content of LDH was measured by a LDH Kit. Results were presented as mean ± SD. Tukey test was used to assess comparisons between individual groups. *P < 0.05, **P < 0.01
Fig. 2
Fig. 2
Effects of DHA-PC on angiogenesis. (A) The wound healing assay. (B) The quantification of the area healing. (C) The Trans-well co-culture model. (D) The Trans-well migration assay. (E) The quantification of the Trans-well migration assay. (F) The networks of tubes were captured under inverted microscope. Scale bar, 100 μm. (G) The disc and surrounding CAMs were photographed under a stereomicroscope. Scale bar, 1 mm. Tukey test was adopted to assess the comparisons between individual groups. *P < 0.05, **P < 0.01
Fig. 3
Fig. 3
Effects of DHA-PC on PPARγ level and key factors level in VEGFR2/Ras/ERK pathway. (A) The mRNA expression of PPARγ. (B) The protein expression of PPARγ. (C) Quantification of protein of PPARγ. (D) The mRNA expression of VEGFR2, Ras, Raf, MEK, and ERK. (E) The protein expression of ERK and p-ERK. (F) Quantification of protein of ERK and p-ERK. Tukey test was adopted to assess the comparisons between individual groups in (A) (D), Student's t test was adopted to assess the comparisons between individual groups in (C) (F). *P < 0.05, **P < 0.01
Fig. 4
Fig. 4
Effects of DHA-PC on HUVECs mRNA and/or protein levels of VE-cadherin and β-catenin. (A) The mRNA expression of VE-cadherin and β-catenin. Tukey test was used to access comparison between individual groups. *P < 0.05, **P < 0.01. (B) The protein level of β-catenin. (C) Quantification of protein of β-catenin. Student's t test was adopted to access the comparison between individual groups. *P < 0.05, **P < 0.01
Fig. 5
Fig. 5
PPARγ antagonist GW9662 reversed the effects that DHA-PC inhibited tubes formation of HUVECs and neovascularization of CAM. (A) The disc and surrounding CAMs were photographed under a stereomicroscope. Scale bar, 1 mm. (B) The networks of tubes were photographed using an inverted microscope. Scale bar, 100 μm. (C) Summary for the mechanisms by DHA-PC mediated anti-angiogenic effects in HUVECs
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Aljada A, O'Connor L, Fu YY, Mousa SA. PPAR gamma ligands, rosiglitazone and pioglitazone, inhibit bFGF- and VEGF-mediated angiogenesis. Angiogenesis. 2008;11:361–367. - PubMed
    1. Berger J, Moller DE. The mechanisms of action of PPARs. Annual Review of Medicine. 2002;53:409–435. - PubMed
    1. Carmeliet P, Jain RK. Molecular mechanisms and clinical applications of angiogenesis. Nature. 2011;473:298–307. - PMC - PubMed
    1. Corton JC, Anderson SP, Stauber A. Central role of peroxisome proliferator-activated receptors in the actions of peroxisome proliferators. Annual Review of Pharmacology and Toxicology. 2000;40:491–518. - PubMed
    1. Ding S, Merkulova-Rainon T, Han ZC, Tobelem G. HGF receptor up-regulation contributes to the angiogenic phenotype of human endothelial cells and promotes angiogenesis in vitro. Blood. 2003;101:4816–4822. - PubMed

LinkOut - more resources