ICAM-1单抗介导的肺血管内皮细胞靶向辛伐他汀纳米载体对急性肺损伤的保护作用与机制研究

Acute lung injury (ALI) is a disease without safe and effective drug intervention methods till now. Pulmonary vascular endothelial cell injury is the key pathophysiology for ALI. Statins have demonstrated their abilities to reverse the process of vascular endothelial cell injury, but the problems needing to further study are uncertain drug efficacy and adverse drug reactions when ALI treated with statins. In our previous study, ICAM-1 monoclonal antibody (anti-ICAM) nanocarriers which were possibly targeting to ALI vascular endothelial cell were prepared, and demonstrated with high drug entrapment efficiency for stains. From these results，we can infer that stains can be successfully targeted to ALI vascular endothelial cell mediated by anti-ICAM nanocarriers. In present study, anti-ICAM stain nanocarriers were continually prepared, and there physicochemical properties and drug levels in targeting cells and tissue were determined at first. Secondly, ALI vascular endothelial cells and animal models were established, and the effect of anti-ICAM stain nanocarriers on ALI including the factors cell proliferation, inflammatory factors, etc, were evaluated thereafter. Finally, the key mechanism for anti-ICAM-1 nanocarrier transporting statin into target cells, which might be related with CAM- mediated passway, were studied. In all, this study may provide a safe, efficient and new stain targeting therapy method for ALI.

急性肺损伤（ALI）迄今尚无安全有效的药物干预手段。肺血管内皮细胞损伤是ALI的关键病理生理机制，他汀类具有逆转血管内皮细胞损伤的作用，但用于治疗ALI仍受限于疗效不确定、不良反应多等问题，亟待研究进一步解决。课题组前期构建的ICAM-1单抗纳米载体，初步证明具有ALI血管内皮细胞的靶向作用，并可高效包载他汀类，提示通过该纳米载体可将他汀类靶向ALI血管内皮细胞。基于此，课题组继续构建ICAM-1单抗他汀纳米载体，研究其理化性质和药物靶细胞和靶组织分布。进一步建立ALI血管内皮细胞和动物模型，从细胞增殖、炎症因子等方面评价ICAM-1单抗他汀纳米载体对ALI的改善作用。最后通过研究ICAM-1单抗纳米载体转运他汀类进入靶细胞与CAM-介导机制的相关性，明确纳米载体靶向转运药物的关键机制。本课题的开展旨在为建立安全、高效的ALI 他汀靶向治疗新方案提供直接的科学依据。

急性肺损伤（ALI）是一种临床常见危重症，迄今并无有效的治疗手段。本研究采用辛伐他汀（SV）为模型药物，构建了一种针对肺血管内皮细胞靶向的ICAM-1单克隆抗体修饰纳米结构脂质载体（NLCs）。 . 采用水性溶剂扩散法，首先制备不同粒径的空白或载药NLCs，NLCs分散液Zeta电位在-20 mV~-30 mV之间，透射电镜显示载药NLCs呈类球型，分布均一。随着载药NLCs粒径增大，药物包封率由66.7%提高至91.04%，体外释放速率减慢。粒径不同的空白NLCs对EAhy926细胞的半数抑制率IC50值介于400-550 μg/mL之间，表明载体具有较低的细胞毒性。细胞摄取结果表明载药NLCs对EAhy926细胞具有良好的内化能力。体内分布结果表明，大粒径NLCs具有显著的肺分布优势。 . 采用合成单氨基终端的聚乙二醇硬脂酸酯（NH2-PEG-SA）作为脂质材料，制备ICAM-1单抗体修饰的辛伐他汀纳米结构脂质载体（anti-ICAM/SV/NLCs）。Anti-ICAM/SV/NLCs粒径约354.7 nm，电位约-32.1 mV，包封率为96.78%，载药量为4.85%，体外药物释放可持续36 h。经LPS诱导ICAM-1表达的EAhy926细胞对anti-ICAM/SV/NLCs的摄取高于非靶向SV/NLCs-3；阻断ICAM-1受体后，EAhy926对anti-ICAM/SV/NLCs的摄取降低。体内分布结果显示，anti-ICAM/SV/NLCs在模型小鼠肺内分布高于对照小鼠及假手术小鼠；且anti-ICAM/SV/NLCs在模型小鼠肺内分布高于非靶向SV/NLCs-3。. 体内药效结果表明，anti-ICAM/SV/NLCs给药24 h及48 h后，可有效改善模型小鼠肺内炎性因子TNF-α、IL-6水平及炎性细胞的浸润。相对于辛伐他汀及SV/NLCs-3干预治疗组，anti-ICAM/SV/NLCs给药后48 h后表现出更显著的治疗优势。HE染色观察小鼠肺组织病理形态学结果表明，给药24 h后anti-ICAM/SV/NLCs干预组小鼠较其他治疗组肺内炎性细胞浸润明显减少，肺间质充血及肺泡隔增厚等病理特征显著改善。本研究显示，针对ALI病理肺组织具有良好靶向特性及改善功效的药物递送系统anti-ICAM/SV/NLCs被成功构建。

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