| 摘要: |
| ［摘要］ 目的 探索人脂肪间充质干细胞来源外泌体（Exosome-HADSCs）联合黄芪多糖（APS）对心肌损伤的保护作用机制。方法 体外培养人脂肪间充质干细胞（HADSCs），提取外泌体，并通过电镜、Western blot法对外泌体进行鉴定。实验设置对照组（control组）、Exo组、APS组、Exo+APS组，采用血管形成实验和划痕实验分析Exosome-HADSCs联合APS对人脐静脉内皮细胞（HUVECs）血管形成和细胞迁移的影响。在AC16心肌细胞损伤模型中，实验设置对照组（control组）、氯化钴（CoCl2）组、CoCl2+Exo组、CoCl2+APS组、CoCl2+Exo+APS组，比较各组细胞存活率、凋亡率及凋亡相关蛋白的表达水平。结果 与control组相比，Exo+APS组HUVECs形成血管网格的数量显著增多（P<0.05），且Exo+APS组形成血管网格的数量多于Exo组和APS组，差异有统计学意义（P<0.05）。与control组相比，Exo+APS组细胞迁移率增高，且Exo+APS组细胞迁移率高于Exo组和APS组，差异有统计学意义（P<0.05）。Exosome-HADSCs联合APS能够显著提高CoCl2损伤AC16细胞的存活率，降低细胞凋亡率，且作用效果较单独使用Exosome-HADSCs和APS更显著（P<0.05）。与CoCl2组相比，CoCl2+Exo+APS组凋亡相关蛋白B淋巴细胞瘤-2相关x蛋白（Bax）、半胱氨酸天冬氨酸蛋白酶-3（caspase-3）表达水平显著降低（P<0.05），抗凋亡蛋白B淋巴细胞瘤-2（Bcl-2）表达水平显著增高（P<0.05），磷脂酰肌醇-3-激酶（PI3K）、磷酸化磷脂酰肌醇-3-激酶（p-PI3K）、苏氨酸-丝氨酸蛋白激酶（Akt）蛋白表达水平均显著增高（P<0.05）。结论 Exosome-HADSCs联合APS对心肌损伤具有保护作用，其机制可能与促进血管形成和内皮细胞迁移，提高细胞存活率，减少心肌细胞凋亡有关，并通过PI3K/Akt信号通路实现。 |
| 关键词: 人脂肪间充质干细胞来源外泌体 黄芪多糖 氯化钴 心肌细胞 磷脂酰肌醇3-激酶/苏氨酸-丝氨酸蛋白激酶信号通路 |
| DOI：10.3969/j.issn.1674-3806.2023.12.09 |
| 分类号:R 542.2 |
| 基金项目:河南省医学科技攻关计划联合共建项目（编号：LHGJ20210853，LHGJ20220957） |
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| A study on the protective mechanism of human adipose mesenchymal stem cell-derived exosomes combined with astragalus polysaccharides against myocardial injury |
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GUO Cai-ru, LIU Xin-xin, NIU Yu-jie, et al.
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Henan International Joint Laboratory of Tumor Cell Immunity and Regenerative Medicine, Luoyang Central Hospital Affiliated to Zhengzhou University, Luoyang 471009, China
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| Abstract: |
| ［Abstract］ Objective To explore the protective mechanism of human adipose mesenchymal stem cell-derived exosomes(Exosome-HADSCs) combined with astragalus polysaccharides(APS) against myocardial injury. Methods Human adipose mesenchymal stem cells(HADSCs) were cultured in vitro, and exosomes were extracted and identified using electron microscopy and Western blot. Control group, Exo group, APS group and Exo+APS group were set up. The effects of Exosome-HADSCs combined with APS on vascular formation and cell migration of human umbilical vein endothelial cells(HUVECs) were analyzed using the angiogenesis test and the scratch test. In the AC16 cardiomyocyte injury model, control group, cobalt chloride(CoCl2) group, CoCl2+Exo group, CoCl2+APS group and CoCl2+Exo+APS group were set up to compare the cell survival rate, apoptosis rate and the expression levels of apoptosis-related proteins among all the groups. Results Compared with that in the control group, the number of vascular mesh formation in HUVECs in the Exo+APS group was significantly increased(P<0.05), and the number of vascular mesh formation in the Exo+APS group was larger than that in the Exo group and the APS group, and the differences were statistically significant(P<0.05). Compared with that in the control group, the cell migration rate in the Exo+APS group was increased, and the cell migration rate in the Exo+APS group was higher than that in the Exo group and the APS group,and the differences were statistically significant(P<0.05). Exosome-HADSCs combined with APS could significantly improve the survival rate of AC16 cells damaged by CoCl2 and decrease the apoptosis rate of AC16 cells damaged by CoCl2, and the effects of Exosome-HADSCs combined with APS were more significant than the effects of Exosome-HADSCs and the effects of APS alone(P<0.05). Compared with those in the CoCl2 group, the expressions of apoptosis-related proteins B-cell lymphoma-2 associated x protein(Bax) and cysteine aspartate specific protease-3(caspase-3) in the CoCl2+Exo+APS group were significantly decreased(P<0.05), and the expression of anti-apoptotic protein B-cell lymphoma-2(Bcl-2) was significantly increased(P<0.05), and the expression levels of phosphatidylinositol-3-kinase(PI3K), phosphated phosphatidylinositol-3-kinase(p-PI3K) and serine-threonine kinase(Akt) were significantly increased(P<0.05). Conclusion Exosome-HADSCs combined with APS have protective effects on myocardial injury, which may be related to promoting angiogenesis and endothelial cell migration, improving cell survival rate, and reducing myocardial cell apoptosis through PI3K/Akt signaling pathway. |
| Key words: Human adipose mesenchymal stem cell-derived exosomes(Exosome-HADSCs) Astragalus polysaccharides(APS) Cobalt chloride(CoCl2) Myocardial cell Phosphatidylinositol-3-kinase(PI3K)/serine-threonine kinase(Akt) signaling pathway |
