| 摘要: |
| 目的 探讨低强度脉冲超声（LIPUS）联合脂肪间充质干细胞（ADSCs）源性外泌体（ADSCs-Exos）对内皮细胞血管生成能力的影响。方法 取SD大鼠腹股沟处脂肪，采用酶消化法分离培养得ADSCs，并对ADSCs表面分子进行鉴定。收集第3代ADSCs上清液提取外泌体，通过透射电子显微镜（TEM）、纳米粒子跟踪分析（NTA）、蛋白免疫印迹技术（Western blot）鉴定。将人脐静脉内皮细胞（HUVECs）按照不同处理方式分为：对照组、外泌体组、LIPUS组及LIPUS+外泌体组，采用CCK-8实验、划痕实验及小管形成实验检测不同处理对HUVECs增殖、迁移及成管能力的影响。。结果 ADSCs-Exos为形态一致的茶托样囊泡，表达标志性蛋白CD9和Tsg101，所测样品直径97.8%集中在118.8±80.8nm，证明提取的样品直径符合外泌体大小。（此特征表现为什么？）。CCK-8实验显示LIPUS+外泌体组OD值为1.882±0.138，均明显高于外泌体组、LIPUS组及对照组（0.646±0.034、0.631±0.027、0.462±0.036），差异均有统计学意义（均P＜0.05）。划痕实验显示LIPUS+外泌体组细胞迁移率为（87.7±4.5）%，均明显高于外泌体组（43.0±5.1）%、LIPUS组（34.7±2.3）%及对照组（25.1±2.0）%，差异均有统计学意义（均P＜0.05）。小管形成实验 显示LIPUS+外泌体组HUVECs成管能力均高于其他组，差异均有统计学意义（均P＜0.05）。结论 LIPUS联合ADSCs-Exos可有效促进血管内皮细胞的增殖、迁移及成管，增强血管生成能力。 |
| 关键词: 脂肪间充质干细胞 外泌体 低强度脉冲超声 血管生成 |
| DOI： |
| 投稿时间：2021-07-01修订日期：2021-11-23 |
| 基金项目:国家自然科学基金资助项目(编号：81901759,81971624) |
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| Low-intensity pulsed ultrasound combined with adipose mesenchymal stem cells derived exosomes enhance the angiogenesis of endothelial cells |
| LIU Junbi,JIANG Riyue,Wang Hao,DENG Qing,ZHOU Qing |
| (Department of Ultrasound Imaging, Renmin Hospital of Wuhan University) |
| Abstract: |
| Objective To explore the enhancement effect of low-intensity pulsed ultrasound （LIPUS） combined with adipose mesenchymal stem cells derived exosomes （ADSCs-Exos） on endothelial angiogenesis.Methods Adipose tissue from SD rats’ groins was obtained. The ADSCs were harvested by enzyme digestion and identified by the surface molecule.The ADSC-Exos were extracted from the supernatant of the 3rd generation ADSCs and the exosome was identified by transmission electron microscopy, Western blot and nanoparticle tracking and analysis technology.Human umbilical vein endothelial cells （HUVECs） were divided into control group，exosome group，LIPUS group and LIPUS+ exosome group，the cell counting kit 8 （CCK-8） assay，scratch test and tubule formation test were used to detect the effects of different treatments on the proliferation，migration and tube formation ability of HUVECs.Results ADSC-Exos were saucer shaped vesicle with uniform morphology, and expressed the membrane-labeled proteins CD9 and Tsg101, 97.8 % of the particle size was 118.8±80.8 nm, which proves that the particle diameter was consistent with exosome size.The CCK-8 analysis showed that LIPUS+exosome group had the best enhancement of HUVECs proliferation compared with exosome group，LIPUS group and control group （OD value 1.882±0.138，0.646±0.034，0.631±0.027 and 0.462±0.036，P＜0.05）.The scratch test showed that the cell mobility of the LIPUS+exosome group was the highest compared with exosome group，LIPUS group and control group （migration rate were 87.7±4.5%，43.0±5.1%，34.7±2.3% and 25.1±2.0%，respectively.P＜0.05）.The tube-forming ability of HUVECs in the LIPUS+exosome group was higher than that of other groups，and the difference was statistically significant （P＜0.05）.Conclusion LIPUS combined with ADSCs-Exos could effectively promote the proliferation，migration and tube formation of HUVECs. |
| Key words: Adipose mesenchymal stem cells Exosomes Low-intensity pulsed ultrasound Angiogenesis |
