胆固醇外排增效型盘状高密度脂蛋白的构建及其抗动脉粥样硬化疗效评价

Construction and Efficacy Evaluation of Cholesterol-efflux Enhanced Discoidal High-density Lipoprotein for Anti-atherosclerosis

关键词:  动脉粥样硬化  逆转动脉粥样硬化斑块  高密度脂蛋白  鞘磷脂  胆固醇逆转运  心血管疾病

Key words:  atherosclerosis  atherosclerosis plaque reversal  high-density lipoprotein  Sphingomyelin  reverse cholesterol transport  cardiovascular disease

摘要:

目的 围绕动脉粥样硬化(Atherosclerosis,AS)斑块中过量的胆固醇蓄积问题,以新生盘状高密度脂蛋白(pre-βHDL)为基材,引入高胆固醇亲和力鞘磷脂(sphingomyelin,SM),构建HDL模拟物SM/βHDL,并探究其促胆固醇逆转运能力和抗AS疗效。方法 首先通过薄膜分散法制备SM/βHDL制剂并表征。利用细胞模型体外模拟AS斑块与肝脏代谢以考察SM/βHDL的胆固醇逆转运效率。以apoE-/-雄鼠为动物模型,联合高胆固醇高脂饮食建立AS模型小鼠。通过血清脂质检测、油红O染色以及切片染色实验考察SM/βHDL对AS斑块消除的疗效。结果 制备的SM/βHDL粒径为(23.42 ± 1.46) nm,呈圆盘状；与未引入SM的βHDL相比,SM/βHDL具有更高的胆固醇亲和力,亲和力KD值为423 nmol.L-1。SM/βHDL促进了(9.88 ± 0.66)%的胆固醇逆转运,相较于βHDL提升了27.5%,相较于临床III期HDL产品CSL-112提升了56.6%。SM/βHDL显著改善血清脂质成分、降低AS斑块面积、降低血管平滑肌细胞数量以提高斑块稳定性。结论 本研究构建的胆固醇外排增效型SM/βHDL能有效逆转并消退AS斑块,从而为βHDL治疗AS提供了新剂型和新思路。

Abstract:

OBJECTIVE In this study, based on the excessive cholesterol accumulation in atherosclerosis (AS) plaques, sphingomyelin (SM) with high cholesterol affinity was introduced on the substrate of the nascent discoidal high-density lipoprotein (pre-β HDL) to construct the HDL analog SM/βHDL. The research aims to investigate SM/βHDL's cholesterol reverse transport influence and anti-AS therapeutic potential. METHODS Firstly, SM/βHDL was prepared by thin film dispersion method and characterized. Its cholesterol reversal efficiency was examined using an in vitro cell model simulating AS plaque and liver metabolism. ApoE-/- male mice combined with high cholesterol and fat diet were used to establish AS model. The therapeutic effect of SM/βHDL on AS plaque elimination was investigated by photoacoustic imaging, serum lipid detection, oil red O staining and section staining, respectively. RESULTS SM/βHDL diameter was (23.42 ± 1.46) nm, exhibiting discoid morphologies. Compared with unincorporated SM, SM/βHDL had a higher cholesterol affinity with an affinity KD value of 423 nmol·L-1. SM/βHDL enhanced the cholesterol reversal efficiency by (9.88 ± 0.66) %, a 27.5% rise compared to βHDL, and a 56.6% increase over the clinical phase III HDL product CSL-112. The findings indicated that SM/βHDL primarily targeted the lesion site, leading to a significant improvement in serum lipid composition, a reduction in AS plaque area, and a decrease in the number of macrophages and vascular smooth muscle cells, thereby enhancing plaque stability. CONCLUSION This research proposes a cholesterol efflux-enhancing SM/βHDL, which effectively reverses and attenuates AS plaques, thus providing a novel and promising therapeutic strategy for treating AS with βHDL.