环孢菌素A-纳米乳联合干细胞移植治疗急性心肌梗死的疗效

Effecacy of Cyclosporine A-nanoparticles Emulsion Combined with Stem Cell Transplantation Therapy for Acute Myocardial Infarction

目的制备并评估负载环孢菌素A-纳米乳药物(CsA-NP)联合脂肪组织来源干细胞(ASCs)移植治疗小型猪急性心肌梗死(AMI)的疗效并研究其可能的机制。方法 CsA-NP采用高压均质法制备而成。经皮球囊封堵冠状动脉左前降支制备小型猪AMI模型(n=17),随机分为对照组(n=5)、CsA-NP组(n=4)、ASCs组(n=4)和CsA-NP+ASCs组(n=4)4组。AMI后1周经冠状动脉内注入ASCs或生理盐水,分别在干细胞移植前和干细胞移植后8周,以延迟增强核磁共振(DE-MRI)评估心脏结构和功能。病理和免疫组织化学评估移植后心肌和移植细胞情况。结果干细胞移植后8周,DE-MRI显示与ASCs组比较,CsA-NP+ASCs组的左心室射血分数显著提高[(53.6±2.4)%比(48.3±1.8)%,P<0.05],梗死面积显著缩小[(6.2±1.7)cm3比(7.5±0.6)cm3,P<0.05],室壁厚度显著增加(P<0.05)。病理和免疫组织化学结果显示,CsA-NP+ASCs组比ASCs组移植存活细胞增加了近3倍,并表达心肌细胞阳性标记物(cTnT、α-actin阳性);与ASCs组比较,CsA-NP+ASCs组进一步减轻梗死心肌纤维化程度,下调了Caspase-3活性,抑制了心肌细胞凋亡。结论 CsA-NP增强了ASCs移植治疗小型猪AMI疗效,提高了心功能,减轻了左心室重构,可能与CsA-NP抑制细胞凋亡、促进细胞增殖分化有关。

Objective To evaluate the efficacy of cyclosporine A-nanoparticles emulsion （CsA-NP） combined with adipose tissue-derived stern cells （ASCs） transplantation therapy for acute myocardial infarction （AMI） in a miniswine model. Methods CsA-NP emulsion was prepared by the high-pressure homogenization method. Models were performed by coronary angioplasty for percutaneous balloon occlusion of left anterior descend- ing artery （LAD） . A total of 17 miniswines survived after AMI were divided into four groups： control group （n = 5）, CsA-NP group （n=4）, ASCs group （n =4）, and CsA-NP ＋ASCs group （n =4） . ASCs or saline weredelivered by intracoronary injection one week after AMI. Before cell transplantation and 8 weeks after cell transplan- tation, delayed-enhanced magnetic resonance imaging （DE-MRI） was performed to evaluate cardiac function and viability. The infarcted myocardium and implanted ceils were histologically studied. Results Eight weeks after treatment, the left ventricular ejection fraction （LVEF） significantly increased in the CsA-NP ＋ ASCs group when compared with the ASCs group [ （53.6 ±2. 4）% vs. （48.3 ± 1.8）%, P 〈0. 05] ; meanwhile, the infarct size significantly decreased [ （6. 2 ± 1.7） cm3 vs. （7.5 ± 0. 6） cm3, P 〈 O. 05 ] and the thickness of the ven- tricular wall significantly increased （P 〈 0. 05 ） . Histology showed that the number of surviving cells increased nearly by three times in the CsA-NP ＋ ASCs group, and the expressions of the cardiomyocyte specific markers （ cTnT and α-actin） were detected. Histological samples also showed that CsA-NP ＋ ASCs group reduced fibrotic tissue, and down-regnlated the activation of Caspase-3. Conclusion The CsA-NP ＋ ASCs combination therapy can enhance the viability of ASCs by improving LVEF and preventing LV expansion, which may be explained that CsA-NP has the anti-apoptotic effect and can promote the survivals and proliferation of ASCs.