Department of Biological Repositories, Zhongnan Hospital of Wuhan University, Wuhan, China; Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA.
Department of Ophthalmology, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA.
Metabolism. 2022 Sep;134:155266. doi: 10.1016/j.metabol.2022.155266. Epub 2022 Jul 19.
Choroidal neovascularization (CNV) in age-related macular degeneration (AMD) leads to blindness. It has been widely reported that increased intake of ω-3 long-chain polyunsaturated fatty acids (LCPUFA) diets reduce CNV. Of the three major pathways metabolizing ω-3 (and ω-6 LCPUFA), the cyclooxygenase and lipoxygenase pathways generally produce pro-angiogenic metabolites from ω-6 LCPUFA and anti-angiogenic ones from ω-3 LCPUFA. Howevehr, cytochrome P450 oxidase (CPY) 2C produces pro-angiogenic metabolites from both ω-6 and ω-3 LCPUFA. The effects of CYP2J2 products on ocular neovascularization are still unknown. Understanding how each metabolic pathway affects the protective effect of ω-3 LCPUFA on retinal neovascularization may lead to therapeutic interventions.
To investigate the effects of LCPUFA metabolites through CYP2J2 pathway and CYP2J2 regulation on CNV both in vivo and ex vivo.
The impact of CYP2J2 overexpression and inhibition on neovascularization in the laser-induced CNV mouse model was assessed. The plasma levels of CYP2J2 metabolites were measured by liquid chromatography and tandem mass spectroscopy. The choroidal explant sprouting assay was used to investigate the effects of CYP2J2 inhibition and specific LCPUFA CYP2J2 metabolites on angiogenesis ex vivo.
CNV was exacerbated in Tie2-Cre CYP2J2-overexpressing mice and was associated with increased levels of plasma docosahexaenoic acids. Inhibiting CYP2J2 activity with flunarizine decreased CNV in both ω-6 and ω-3 LCPUFA-fed wild-type mice. In Tie2-Cre CYP2J2-overexpressing mice, flunarizine suppressed CNV by 33 % and 36 % in ω-6, ω-3 LCPUFA diets, respectively, and reduced plasma levels of CYP2J2 metabolites. The pro-angiogenic role of CYP2J2 was corroborated in the choroidal explant sprouting assay. Flunarizine attenuated ex vivo choroidal sprouting, and 19,20-EDP, a ω-3 LCPUFA CYP2J2 metabolite, increased sprouting. The combined inhibition of CYP2J2 with flunarizine and CYP2C8 with montelukast further enhanced CNV suppression via tumor necrosis factor-α suppression.
CYP2J2 inhibition augmented the inhibitory effect of ω-3 LCPUFA on CNV. Flunarizine suppressed pathological choroidal angiogenesis, and co-treatment with montelukast inhibiting CYP2C8 further enhanced the effect. CYP2 inhibition might be a viable approach to suppress CNV in AMD.
年龄相关性黄斑变性（AMD）中的脉络膜新生血管（CNV）可导致失明。已有大量研究报道表明，增加 ω-3 长链多不饱和脂肪酸（LCPUFA）的摄入可减少 CNV。在代谢 ω-3（和 ω-6 LCPUFA）的三大途径中，环加氧酶和脂加氧酶途径通常从 ω-6 LCPUFA 产生促血管生成代谢物，从 ω-3 LCPUFA 产生抗血管生成代谢物。然而，细胞色素 P450 氧化酶（CPY）2C 可从 ω-6 和 ω-3 LCPUFA 产生促血管生成代谢物。CYP2J2 产物对眼部新生血管的影响尚不清楚。了解每种代谢途径如何影响 ω-3 LCPUFA 对视网膜新生血管的保护作用，可能会导致治疗干预。
研究 CYP2J2 途径和 CYP2J2 对体内和体外 CNV 的 LCPUFA 代谢物的影响。
评估 CYP2J2 过表达和抑制对激光诱导的 CNV 小鼠模型中新生血管的影响。通过液相色谱-串联质谱法测量 CYP2J2 代谢物的血浆水平。使用脉络膜外植体发芽试验研究 CYP2J2 抑制和特定的 CYP2J2 ω-3 LCPUFA 代谢物对体外血管生成的影响。
Tie2-Cre CYP2J2 过表达小鼠的 CNV 加重，且与血浆二十二碳六烯酸水平升高有关。用氟桂利嗪抑制 CYP2J2 活性可降低 ω-6 和 ω-3 LCPUFA 喂养的野生型小鼠的 CNV。在 Tie2-Cre CYP2J2 过表达小鼠中，氟桂利嗪分别抑制 ω-6 和 ω-3 LCPUFA 饮食中的 CNV 33%和 36%，并降低 CYP2J2 代谢物的血浆水平。CYP2J2 在脉络膜外植体发芽试验中的促血管生成作用得到了证实。氟桂利嗪减弱了体外脉络膜发芽，而 ω-3 LCPUFA 的 CYP2J2 代谢物 19,20-EDP 增加了发芽。用氟桂利嗪联合 montelukast 联合抑制 CYP2C8 进一步通过抑制肿瘤坏死因子-α 来增强 CNV 抑制，从而增强 CYP2J2 的抑制作用。
CYP2J2 抑制增强了 ω-3 LCPUFA 对 CNV 的抑制作用。氟桂利嗪抑制病理性脉络膜血管生成，并用 montelukast 联合抑制 CYP2C8 进一步增强该作用。CYP2 抑制可能是抑制 AMD 中 CNV 的一种可行方法。
