Vara-Messler M, Trevisi L, Zulato E, Ramaschi G E, Risé P, Pinna C, Indraccolo S, Sala A, Bolego C
Department of Pharmaceutical and Pharmacological Sciences, University of Padova, Padova, Italy.
Pole of Pharmacology and Therapeutics (FATH), Institut de Recherche Expérimentale et Clinique (IREC), Université Catholique de Louvain (UC Louvain), Brussels, Belgium.
Front Pharmacol. 2025 Feb 25;16:1524980. doi: 10.3389/fphar.2025.1524980. eCollection 2025.
Blood vessels supply oxygen, nutrients and provide gateways for immune surveillance. Since this network nourishes all tissues, vessel abnormalities contribute to many diseases, such as cancer. One of the potential targets for Docosahexaenoic Acid (DHA) in cancer is suppressing angiogenesis, a process of new blood vessel formation within tumors. In addition, aspirin (ASA) has antineoplastic effects that may be mediated, at least in part, by metabolites derived from acetylated COX-2. We aimed at determining the effect of DHA as well as its metabolites in angiogenesis, using as well as models.
Endothelial cell (EC) proliferation, motility and capillary-like tube formation were determined by MTT, wound healing, Boyden and Matrigel assays, respectively. In vivo angiogenesis was measured by the Matrigel sponge model in mice. The biosynthesis of proresolving lipid mediators by ECs was determined by LC-MS-MS.
DHA, but not arachidonic acid (AA), at concentrations consistent with those reached in blood after fish oil supplementation, decreased EC migration in a time- and concentration-dependent manner. Pretreatment with ASA modulated cell migration already after 24 h, while both DHA and ASA decreased migration at longer incubation times without affecting viability. 17-hydroxy-DHA was detected upon incubation with DHA, and increased amounts were observed upon combined treatment with DHA and ASA, an increase that was associated to a synergic effect on EC migration. 17(R)-hydroxy-DHA (17R-HDHA), the metabolite resulting from acetylated COX-2 activity of DHA, reduced EC migration in a concentration-dependent manner. DHA in the presence of ASA, as well as 17R-HDHA, also reduced EC tube formation. These results were confirmed where both 17R-HDHA or its downstream metabolite 17RResolvinD1 were able to decrease microvessels density in a Matrigel sponge model. Overall, we demonstrated that DHA in the presence of ASA-dependent acetylation of COX-2 showed increased antiangiogenic effects, possibly resulting from its conversion to its hydroxylated derivatives.
血管为组织提供氧气、营养物质，并为免疫监视提供通道。由于该网络滋养所有组织，血管异常会导致许多疾病，如癌症。二十二碳六烯酸（DHA）在癌症中的潜在靶点之一是抑制血管生成，即肿瘤内新血管形成的过程。此外，阿司匹林（ASA）具有抗肿瘤作用，其作用可能至少部分由乙酰化COX-2衍生的代谢产物介导。我们旨在使用[具体模型1]和[具体模型2]模型，确定DHA及其代谢产物在血管生成中的作用。
分别通过MTT法、伤口愈合试验、博伊登试验和基质胶试验测定内皮细胞（EC）的增殖、迁移和毛细血管样管形成。通过小鼠基质胶海绵模型测量体内血管生成。通过液相色谱-质谱-质谱法测定ECs中促分解脂质介质的生物合成。
在与补充鱼油后血液中达到的浓度一致的浓度下，DHA而非花生四烯酸（AA）以时间和浓度依赖性方式降低EC迁移。ASA预处理在24小时后即调节细胞迁移，而DHA和ASA在较长孵育时间均降低迁移，且不影响细胞活力。与DHA孵育后检测到17-羟基-DHA，DHA与ASA联合处理后观察到其含量增加，这种增加与对EC迁移的协同作用相关。17（R）-羟基-DHA（17R-HDHA）是DHA经乙酰化COX-2活性产生的代谢产物，以浓度依赖性方式降低EC迁移。在ASA存在下的DHA以及17R-HDHA也减少EC管形成。在[具体模型]中证实了这些结果，其中17R-HDHA或其下游代谢产物17R-消退素D1均能够在基质胶海绵模型中降低微血管密度。总体而言，我们证明在COX-2依赖的ASA乙酰化存在下，DHA显示出增强的抗血管生成作用，这可能是由于其转化为羟基化衍生物所致。
