5-氨基酮戊酸介导的声动力疗法快速抑制动脉粥样硬化斑块进展

Rapid inhibition of atherosclerotic plaque progression by sonodynamic therapy

目的研究5-氨基酮戊酸(ALA)介导的声动力疗法(SDT)对动脉粥样硬化斑块的影响。方法球囊损伤加高脂喂养建立兔股动脉粥样硬化斑块模型,荧光显微镜及荧光光谱仪检测静脉注射ALA后0、1、2、3、4、5及6 h(n=3),其代谢产物原卟啉Ⅸ(PpⅨ)在斑块中的代谢与分布。动物随机分为对照组及SDT组。SDT治疗后6、12、24及72 h(n=7),TUNEL及免疫组织化学方法检测斑块中细胞凋亡及巨噬细胞含量;治疗后1周,行高频超声、病理及免疫组织化学检测。结果 ALA注射后2 h,PpⅨ在斑块中含量达到高峰并主要分布在巨噬细胞表达阳性区域。与对照组相比,SDT治疗后6、12、24及72 h斑块中细胞凋亡分别增加3.1、3.8、7.5及4.0倍,SDT治疗后24及72 h斑块中巨噬细胞含量减少45%及67%;与治疗前相比,SDT治疗后1周,股动脉直径狭窄率减少14%。与对照组相比,SDT组斑块面积减小38%,管腔面积增加86%,斑块中脂质、巨噬细胞、增殖期细胞、白细胞介素1β(IL-1β)及肿瘤坏死因子α(TNF-α)含量分别减少64%、71%、76%、62%及60%,胶原含量增加117%,差异均有统计学意义(P<0.05)。结论 ALA-PpⅨ主要分布在动脉粥样硬化斑块的巨噬细胞中;SDT诱导斑块内细胞凋亡,改变斑块内成分促进斑块稳定并减小斑块大小。SDT是一种潜在的无创治疗动脉粥样硬化斑块的方法。

Aim To investigate the effect of aminolaevulinic acid (ALA)-mediated sonodynamic therapy (SDT) on atherosclerosis plaque. Methods The rabbit atherosclerotic model was established by balloon denudation and an atherogenic diet. The metabolism and distribution of protoporphyrin Ⅸ(PpⅨ) in the plaque were detected by fluorescence microscopy and fluorescence spectrometry at 0,1, 2,3, 4,5 and 6 hours after intravenous injection of ALA (n=3). Animals were randomly divided into control group and SDT group. Apoptosis and macrophage content were detected by TUNEL and immunohistochemistry at 6,12, 24 and 72 hours (n=7) after SDT treatment, and high-frequency ultrasound, pathology and immunohistochemistry were performed at 1 week after SDT treatment. Results Two hours after ALA injection, the content of PpⅨ in plaques reached a peak and mainly distributed in the areas of macrophages. Compared with the control group, the apoptosis of plaque cells increased by 3.1,3.8,7.5 and 4.0 folds at 6,12, 24 and 72 hours after SDT treatment, and the content of macrophages decreased by 45% and 67% at 24 and 72 hours after SDT treatment, and the stenosis rate of femoral artery diameter decreased by 14% at 1 week after SDT treatment. Compared with the control group, the plaque area of SDT group decreased by 38%;the lumen area increased by 86%;the content of lipid, macrophage, proliferative cells, IL-1 beta and TNF-alpha decreased by 64%, 71%, 76%, 62% and 60% respectively;and the content of collagen increased by 117%. Conclusions ALA-PpⅨ is mainly distributed in intraplaque macrophages, SDT induces macrophages apoptosis, and changes the components of plaque to promote plaque stability and reduce the size of plaque. SDT is a potential noninvasive method for the treatment of atherosclerotic plaques.