Exosomal miRNA-215-5p Derived from Adipose-Derived Stem Cells Attenuates Epithelial-Mesenchymal Transition of Podocytes by Inhibiting ZEB2
Exosomal miRNA-215-5p Derived from Adipose-Derived Stem Cells Attenuates Epithelial-Mesenchymal Transition of Podocytes by Inhibiting ZEB2
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DOI：
10.1155/2020/2685305
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发表时间：
2020-02-22
影响因子：
--
通讯作者：
He, Qiang
中科院分区：
文献类型：
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作者：
Jin, Juan;Wang, Yunguang;He, Qiang
背景足细胞迁移参与足细胞丢失和蛋白尿的产生，与糖尿病肾病（DN）的发生发展密切相关。来自脂肪来源干细胞的外泌体（ADSCs-Exos）在治疗DN中有效地抑制足细胞凋亡。然而，ADSCs-Exos如何影响足细胞的迁移尚不清楚。本研究旨在探讨ADSCs-Exos对细胞迁移的调控作用及其机制。方法.通过透射电子显微镜（TEM）、蛋白质印迹和流式细胞术鉴定ADSCs-Exo。分别通过CCK 8和Transwell测定法测量足细胞的细胞活力和迁移能力。通过qRT-PCR测定miRNA和mRNA的相对表达。IF法检测PKH 26标记的exosome与足细胞之间的传递。采用双荧光素酶报告基因检测miR-215- 5 p与ZEB 2的关系。结果暴露于来自DN患者的血清（hDN-血清）显著抑制足细胞的细胞活力，但ADSCs-Exo添加显著减弱由hDN-血清的瞬时刺激诱导的细胞毒性。此外，ADSCs-Exo给药可有效抑制高糖诱导的足细胞迁移和损伤。在足细胞功能障碍的情况下，HG处理下的几种miRNA呈现出显著下降，包括miR-251- 5 p、miR-879- 5 p、miR-3066- 5 p和miR-7a-5 p，所有这些都被添加ADSCs-Exo所拯救。然而，只有miR-251- 5 p是ADSCs-Exo介导的足细胞损伤保护作用过程中的关键决定因素。miR-251- 5 p抑制剂抵消了ADSCs-Exo制剂对HG诱导的增殖抑制和迁移促进的改善。此外，单独的miR-215- 5 p模拟物显著逆转了HG诱导的足细胞EMT过程。从机制上讲，我们证实ADSCs-Exos介导miR-215- 5 p穿梭至足细胞，从而保护免受HG诱导的转移，这可能是通过抑制ZEB 2的转录。结论ADSCs-Exo通过涉及ZEB 2的机制对HG诱发的足细胞EMT进展具有保护作用。ADSCs-Exo制剂可能是临床上改善足细胞功能障碍和DN症状的有用治疗策略。
Background. Podocyte migration is actively involved in the process of podocyte loss and proteinuria production, which is closely associated with the development of diabetic nephropathy (DN). Exosomes from adipose-derived stem cells (ADSCs-Exos) effectively inhibit podocyte apoptosis in the treatment of DN. However, how ADSCs-Exos affect the migration of podocytes is obscure. This study is aimed at exploring the regulatory role of ADSCs-Exos on cell migration and the underlying mechanism. Methods. ADSCs-Exo was authenticated by transmission electron microscopy (TEM), western blotting, and flow cytometry. Cell viability and migration ability of podocytes were measured by CCK8 and Transwell assays, respectively. Relative expressions of miRNAs and mRNAs were determined by qRT-PCR. The transmitting between PKH26-labeled exosome and podocytes was evaluated by IF assay. Dual luciferase reporter assay was employed to detect the relationship between miR-215-5p and ZEB2. Results. The exposure to serum from DN patient (hDN-serum) significantly inhibited cell viability of podocytes, but ADSCs-Exo addition notably blunts cytotoxicity induced by the transient stimulus of hDN-serum. Besides, ADSCs-Exo administration powerfully impeded high glucose- (HG-) induced migration and injury of podocyte. With the podocyte dysfunction, several miRNAs presented a significant decline under the treatment of HG including miR-251-5p, miR-879-5p, miR-3066-5p, and miR-7a-5p, all of which were rescued by the addition of ADSCs-Exo. However, only miR-251-5p was a key determinant in the process of ADSCs-Exo-mediated protective role on podocyte damage. The miR-251-5p inhibitor counteracted the improvement from the ADSCs-Exo preparation on HG-induced proliferation inhibition and migration promotion. Additionally, miR-215-5p mimics alone remarkably reversed HG-induced EMT process of podocyte. Mechanistically, we confirmed that ADSCs-Exos mediated the shuttling of miR-215-5p to podocyte, thereby protecting against HG-induced metastasis, possibly through inhibiting the transcription of ZEB2. Conclusion. ADSCs-Exo has the protective effect on HG-evoked EMT progression of podocytes thru a mechanism involving ZEB2. Potentially, the ADSCs-Exo preparation is a useful therapeutic strategy for improving podocyte dysfunction and DN symptoms clinically.