Department of Molecular & Cellular Physiology, Louisiana State University Health Shreveport, Shreveport, LA, 71103, USA.
Department of Molecular & Cellular Physiology, Louisiana State University Health Shreveport, Shreveport, LA, 71103, USA.
Exp Eye Res. 2021 May;206:108540. doi: 10.1016/j.exer.2021.108540. Epub 2021 Mar 15.
Central retinal artery occlusion, retinopathy, and retinal neovascularization have been reported in methamphetamine (METH) abusers. In the current study, we investigated whether METH induces retinal neovascularization in a mouse model, and if so, whether the neovascularization is associated with increased hypoxia, hypoxia-inducible factor 1α (HIF-1α), and vascular endothelial growth factor (VEGF). Mice were administrated METH by intraperitoneal injection over a 26-day period, or injected with saline as a vehicle control. The number of retinal arterioles and venules were counted using in vivo live imaging following infusion with fluorescein isothiocyanate-dextran. Excised retinas were stained with griffonia simplicifolia lectin I and flat mounted for a measurement of vascularity (length of vessels per tissue area) with AngioTool. Retinal hypoxia was examined by formation of pimonidazole adducts with an anti-pimonidazole antibody, and HIF-1α and VEGFa protein levels in the retina were detected by immunoblot. METH administration increased vascularity (including the number of arterioles) measured on Day 26. Retinal VEGFa protein level was not changed in METH-treated mice on Day 5, but was increased on Day 12 and Day 26. Hypoxia (pimonidazole adduct formation) was increased in retinas of METH-treated mice on Day 12 and Day 26, as were HIF-1α protein expression levels. These results indicate that METH administration induces hypoxia, HIF-1α, VEGFa, and angiogenesis in the retina.
美沙酮(METH)滥用者已报告出现中央视网膜动脉阻塞、视网膜病变和视网膜新生血管形成。在目前的研究中,我们调查了 METH 是否会在小鼠模型中诱导视网膜新生血管形成,如果是,新生血管形成是否与缺氧增加、缺氧诱导因子 1α(HIF-1α)和血管内皮生长因子(VEGF)有关。小鼠通过腹腔注射在 26 天内给予 METH,或注射生理盐水作为载体对照。用异硫氰酸荧光素-葡聚糖灌注后,通过体内活体成像计数视网膜小动脉和小静脉的数量。用 Griffonia simplicifolia lectin I 对切除的视网膜进行染色,并进行平置,用 AngioTool 测量血管密度(组织面积内的血管长度)。用抗 pimonidazole 抗体检测视网膜缺氧,用免疫印迹检测视网膜中 HIF-1α 和 VEGFa 蛋白水平。METH 给药增加了第 26 天测量的血管密度(包括小动脉数量)。METH 处理的小鼠在第 5 天视网膜 VEGFa 蛋白水平没有变化,但在第 12 天和第 26 天增加。第 12 天和第 26 天,METH 处理的小鼠视网膜缺氧(pimonidazole 加合物形成)和 HIF-1α 蛋白表达水平增加。这些结果表明,METH 给药诱导视网膜缺氧、HIF-1α、VEGFa 和血管生成。