Yuan Hong, Hu Haiqiang, Sun Jindong, Shi Mingjuan, Yu Huamin, Li Cairong, Sun Y U, Yang Zhijian, Hoffman Robert M
Yuhang District First People's Hospital, Hangzhou, P.R. China
Yuhang District First People's Hospital, Hangzhou, P.R. China.
In Vivo. 2018 Sep-Oct;32(5):1025-1032. doi: 10.21873/invivo.11342.
BACKGROUND/AIM: Intraplaque neovascularization is often associated with plaque formation, development and instability, and clinical symptoms in atherosclerosis. The aim of the present study was to investigate a new strategy for treating athrosclerosis by ultrasound-targeted microbubble delivery (UTMD) targeting intraplaque neovascularization in an APOE-deficient mouse model of atherosclerosis.
A mouse model of atherosclerosis was induced by feeding Apoe mice a hypercholesterolemic diet and was verified with hematoxylin and eosin staining and intercellular adhesion molecule 1 (ICAM-1) expression. Targeted microbubbles (MB) were prepared by conjugating microbubbles with biotinylated antibody to ICAM1 (MBi) or with both biotinylated anti-ICAM1 and the angiogenesis inhibitor Endostar (MBie). The targeted microbubbles were analyzed with epifluorescence microscopy and flow cytometry. The animals with induced atherosclerotic plaques received MBi or MBie followed by UTMD treatment. Endostar treatment alone was given to other animals for comparison. Morphological assessment of atherosclerotic plaques was performed after treatment. The expression of angiogenesis marker CD31 was detected by immunohistochemical analysis.
Atherosclerotic plaques developed in the entire aorta with significant intraplaque ICAM-1 expression in the APOE-deficient mice following a 30-week hypercholesterolemic diet. Microbubbles were successfully conjugated with anti-ICAM-1 and Endostar, with a conjugation rate of 98.3% and 63.5%, respectively. UTMD with MBie significantly reduced the area of atherosclerotic plaque as compared to the model control (p<0.05). Treatment with Endostar and UTMD with MBie significantly reduced CD31 expression compared with the model control group (p<0.01). Greater significant inhibitory effect on CD31 expression was found in the group treated with UTMD and MBie compared to the Endostar- and UTMD with MBi groups (p<0.01).
UTMD targeting intraplaque neovascularization was found to inhibit atherosclerotic plaque in a mouse model of atherosclerosis, suggesting the potential of microbubble-mediated ultrasound technology in aiding drug delivery for atherosclerosis treatment.
背景/目的：斑块内新生血管形成常与动脉粥样硬化中的斑块形成、发展和不稳定性以及临床症状相关。本研究的目的是在载脂蛋白E缺陷型动脉粥样硬化小鼠模型中，研究一种通过超声靶向微泡给药（UTMD）靶向斑块内新生血管形成来治疗动脉粥样硬化的新策略。
通过给载脂蛋白E基因敲除小鼠喂食高胆固醇饮食诱导动脉粥样硬化小鼠模型，并用苏木精-伊红染色和细胞间黏附分子1（ICAM-1）表达进行验证。通过将微泡与抗ICAM1生物素化抗体（MBi）或抗ICAM1生物素化抗体和血管生成抑制剂恩度（Endostar）两者偶联来制备靶向微泡（MB）。用落射荧光显微镜和流式细胞术分析靶向微泡。对诱导出动脉粥样硬化斑块的动物给予MBi或MBie，随后进行UTMD治疗。对其他动物单独给予恩度治疗作为对照。治疗后对动脉粥样硬化斑块进行形态学评估。通过免疫组织化学分析检测血管生成标志物CD31的表达。
在30周高胆固醇饮食后，载脂蛋白E缺陷型小鼠的整个主动脉中均出现动脉粥样硬化斑块，且斑块内ICAM-1表达显著。微泡成功与抗ICAM-1和恩度偶联，偶联率分别为98.3%和63.5%。与模型对照组相比，MBie的UTMD显著减小了动脉粥样硬化斑块面积（p<0.05）。与模型对照组相比，恩度治疗组以及MBie的UTMD治疗组显著降低了CD31表达（p<0.01）。与恩度治疗组以及MBi的UTMD治疗组相比，MBie的UTMD治疗组对CD31表达的抑制作用更显著（p<0.01）。
在动脉粥样硬化小鼠模型中发现，靶向斑块内新生血管形成的UTMD可抑制动脉粥样硬化斑块，提示微泡介导的超声技术在辅助药物递送治疗动脉粥样硬化方面具有潜力。
