Yuan Wanzhong, Zhang Jianlin, Huo Ran, Hou Chaofan, Yang Jun, Wang Tao
Department of Neurosurgery, Peking University Third Hospital, Beijing 100191, China.
Center of Basic Medical Research, Institute of Medical Innovation and Research, Peking University Third Hospital, Beijing 100191, China.
J Cardiovasc Dev Dis. 2023 Jul 19;10(7):309. doi: 10.3390/jcdd10070309.
Iron overload can accelerate the accumulation of lipid oxides and contribute to the progression of atherosclerosis. Ferritin heavy chain (FT-H) exhibits oxidase activity, which inhibits the toxicity of ferrous ions and reduces oxidative damage. We investigated the effect of the intraperitoneal injection of FT-H on the progression of atherosclerosis in APOE-knockout mice (Apo-E mice). All mice were fed on a high-fat diet. After 10 weeks, the mice were divided into an injection group ( = 4) and a control group ( = 4). The injection group was injected intraperitoneally with FT-H (50 mg/kg, once a week), and the control group was treated with PBS buffer (at an equal volume to the injection group, once a week). After 10 weeks of intervention, MRI of the aortas was performed. Then, the animals were sacrificed, and tissues were taken. Hematoxylin-eosin (HE) staining was used for histomorphometry, Masson staining was used to quantify the collagen content in the arteries, Prussian blue staining was used to visualize iron deposition in the arteries, and MRI was used to analyze the structure of the aorta in vivo. Immunohistochemistry was performed to detect the expression of MCP-1, MMP-2, MMP-9, FT-H, FT-L, TfR1, NRF-2 and GPX-4. The serological results showed that the injection group had lower levels of glucose (Glu), triacylglycerol (TG), cholesterol (CHO), low-density lipoprotein-C (LDL-C) and malondialdehyde (MDA) ( = 0.0058, = 0.0098, = 0.0019, = 0.0368 and = 0.0025, respectively), and their serum ferritin (SF) and superoxide dismutase (SOD) levels were higher ( = 0.0004 and < 0.0001). The Masson staining and MRI results showed that the injection group had less collagen deposition ( = 0.0226), a larger arterial lumen area and arterial volume ( = 0.0006 and = 0.0005), thinner arterial wall thickness ( = 0.0013) and a more stable arterial plaque structure ( < 0.0001). The immunohistochemical results showed reduced expression of FT-H, FT-L, TfR1, MMP-2, MMP-9, MCP-1 and NRF-2 in the injection group ( = 0.0054, = 0.0242, = 0.0221, = 0.0477, = 0.0131, = 0.0435 and = 0.0179). Prussian blue staining showed that the area of iron-positive areas in the aortic plaques of the control group was larger than that of injected group. The expression of GPX-4 was lower in the control group than in the injection group ( = 0.016). The intraperitoneal administration of FT-H to Apo-E mice resulted in lower blood glucose and lipid levels; reduced iron and iron metabolism protein deposition in the aorta; reduced indices of their ferroptosis, oxidation and inflammatory aggregation; and reduced collagen deposition in the aorta, which delayed the process of aortic atherosclerosis in mice.
铁过载可加速脂质氧化物的积累并促进动脉粥样硬化的进展。铁蛋白重链（FT-H）具有氧化酶活性，可抑制亚铁离子的毒性并减少氧化损伤。我们研究了腹腔注射FT-H对载脂蛋白E基因敲除小鼠（Apo-E小鼠）动脉粥样硬化进展的影响。所有小鼠均喂食高脂饮食。10周后，将小鼠分为注射组（n = 4）和对照组（n = 4）。注射组腹腔注射FT-H（50 mg/kg，每周一次），对照组用PBS缓冲液处理（体积与注射组相同，每周一次）。干预10周后，对主动脉进行磁共振成像（MRI）检查。然后，处死动物并取材。苏木精-伊红（HE）染色用于组织形态计量学分析，Masson染色用于定量动脉中的胶原含量，普鲁士蓝染色用于观察动脉中的铁沉积，MRI用于在体分析主动脉结构。进行免疫组织化学检测单核细胞趋化蛋白-1（MCP-1）、基质金属蛋白酶-2（MMP-2）、基质金属蛋白酶-9（MMP-9）、FT-H、铁蛋白轻链（FT-L）、转铁蛋白受体1（TfR1）、核因子E2相关因子2（NRF-2）和谷胱甘肽过氧化物酶4（GPX-4）的表达。血清学结果显示，注射组的葡萄糖（Glu）、三酰甘油（TG）、胆固醇（CHO）、低密度脂蛋白胆固醇（LDL-C）和丙二醛（MDA）水平较低（分别为P = 0.0058、P = 0.0098、P = 0.0019、P = 0.0368和P = 0.0025），其血清铁蛋白（SF）和超氧化物歧化酶（SOD）水平较高（P = 0.0004和P < 0.0001）。Masson染色和MRI结果显示，注射组的胶原沉积较少（P = 0.0226），动脉管腔面积和动脉体积较大（P = 0.0006和P = 0.0005），动脉壁厚度较薄（P = 0.0013），动脉斑块结构更稳定（P < 0.0001）。免疫组织化学结果显示，注射组中FT-H、FT-L、TfR1、MMP-2、MMP-9、MCP-1和NRF-2的表达降低（P = 0.0054、P = 0.0242、P = 0.0221、P = 0.0477、P = 0.0131、P = 0.0435和P = 0.0179）。普鲁士蓝染色显示，对照组主动脉斑块中铁阳性区域的面积大于注射组。对照组中GPX-4的表达低于注射组（P = 0.016）。对Apo-E小鼠腹腔注射FT-H可降低血糖和血脂水平；减少主动脉中铁和铁代谢蛋白的沉积；降低其铁死亡、氧化和炎症聚集指标；减少主动脉中的胶原沉积，从而延缓小鼠主动脉粥样硬化进程。
