| 引用本文： |
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金翠,曹永梅,尚嘉伟,等.骨髓间充质干细胞来源外泌体保护脓毒症相关急性肾损伤的体外研究[J].同济大学学报（医学版）,2022,43(2):157-164. [点击复制]
- JIN Cui,CAO Yongmei,SHANG Jiawei,et al.Protective effects of exosomes derived from bone mesenchymal stem cells in sepsis-induced acute kidney injury cell model in vitro[J].同济大学学报（医学版）,2022,43(2):157-164. [点击复制]
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| 摘要: |
| 目的研究骨髓间充质干细胞来源外泌体（exosomes derived from bone mesenchymal stem cells, BMSCs-Exo）对脓毒症相关急性肾损伤体外模型的作用。方法利用超速离心法提取骨髓间充质干细胞分泌的外泌体，通过透射电镜、纳米颗粒跟踪分析及Western印迹法鉴定外泌体。用脂多糖（lipopolysaccharide, LPS）处理人肾小管上皮细胞（HK-2）建立脓毒症相关急性肾损伤（sepsis-induced acute kidney injury, SAKI）细胞模型，将细胞分为对照组（CON组）、脂多糖组（LPS组）和脂多糖+外泌体组（LPS+EXO组）。CCK-8法检测各组细胞活性、流式细胞仪检测细胞凋亡率，应用Western印迹法检测细胞凋亡相关蛋白cleaved-Caspase3、Bax和bcl2水平，采用qRT-PCR检测炎性因子TNF-α、IL-6和IL-1β mRNA水平。结果应用LPS处理HK-2细胞成功构建了SAKI细胞模型。LPS组中，HK-2细胞经LPS（100 μg/mL）处理后细胞活性和抗凋亡蛋白bcl2表达明显下降（P<0.05），而细胞凋亡率和凋亡蛋白cleaved-Caspase3、Bax表达水平显著上升（P<0.01），同时炎性指标TNF-α、IL-6和IL-1β mRNA表达上调（P<0.01）。LPS+EXO组中，BMSCs-Exo逆转了LPS对HK-2细胞的作用，细胞活性升高、凋亡蛋白cleaved-Caspase3以及Bax下降和抗凋亡蛋白bcl2升高（P<0.05），同时还下调了炎性因子（TNF-α、IL-6和IL-1β）mRNA表达水平（P<0.05）。结论BMSCs-Exo在LPS诱导的SAKI体外模型中发挥抗炎和抗凋亡的保护作用，为未来临床利用BMSCs-Exo治疗SAKI提供理论基础。 |
| 关键词: 骨髓间充质干细胞 外泌体 脓毒症 急性肾损伤 |
| DOI：10.12289/j.issn.1008-0392.21396 |
| 投稿时间：2021-09-18 |
| 基金项目:上海市公共卫生体系建设三年行动计划（GWV-10.1-XK23）；上海市2021年度“科技创新行动计划”医学创新研究专项（21Y11902800） |
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| Protective effects of exosomes derived from bone mesenchymal stem cells in sepsis-induced acute kidney injury cell model in vitro |
| JIN Cui,CAO Yongmei,SHANG Jiawei,LI Yingchuan |
| (Department of Critical Care Medicine, Shanghai Jiao Tong University Affiliated Sixth People’s Hospital, Shanghai 200233, China) |
| Abstract: |
| ObjectiveTo investigate the effects of exosomes derived from bone mesenchymal stem cells(BMSCs-Exo) in a sepsis-induced acute kidney injury(SAKI) cell model. MethodsBMSCs-Exo were isolated by differential ultracentrifugation. Transmission electron microscope(TEM), nanoparticle tracking analysis(NTA) and Western blot were used to identify exosomes. The in vitro model of SAKI was constructed by treating HK-2 cells with LPS, then the cells were divided into three groups(CON, LPS and LPS+EXO). Cell viability was assessed by CCK-8. Apoptosis rate was measured by flow cytometry. Protein levels of apoptosis-associated protein cleaved-Caspase3, Bax and bcl2 were tested by Western blot. The mRNA expression levels of TNF-α, IL-6 and IL-1β were detected by qRT-PCR. ResultsLPS treatment decreased the cell viability and the expression of anti-apoptosis protein bcl2（P<0.05）, elevated the apoptosis rate and the expressions of apoptosis-related protein cleaved-Caspase3 and Bax（P<0.01）. Meanwhile, the mRNA expressions of TNF-α, IL-6 and IL-1β were increased（P<0.01）. However, BMSCs-Exo was able to reverse LPS-induced decreased cell viability, increased levels of apoptosis and inflammatory response（P<0.05）. ConclusionBMSCs-Exos can exert anti-apoptosis and anti-inflammatory effects in LPS-treated HK-2 cells, indicating its potential clinical application for treatment of SAKI. |
| Key words: bone mesenchymal stem cells exosomes sepsis acute kidney injury |
