Pre-incubation with hucMSC-exosomes prevents cisplatin-induced nephrotoxicity by activating autophagy.
Pre-incubation with hucMSC-exosomes prevents cisplatin-induced nephrotoxicity by activating autophagy.
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DOI：
10.1186/s13287-016-0463-4
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发表时间：
2017-04-08
影响因子：
7.5
通讯作者：
Xu W
中科院分区：
文献类型：
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作者：
Wang B;Jia H;Zhang B;Wang J;Ji C;Zhu X;Yan Y;Yin L;Yu J;Qian H;Xu W
由于顺铂的肾毒性副作用，给药是有限的，而且很难预防顺铂的这种肾毒性。间充质干细胞(MSC)来源的外切体已被认为是一种治疗组织损伤的新方法。在本研究中，我们在体内外证实了人脐血间充质干细胞来源的外切体(hucMSC-Ex)的预处理可以通过激活自噬来预防顺铂引起的肾毒性。体外实验中，大鼠肾小管上皮细胞(NRK-52E)分别与人肺成纤维细胞(HucMSC)或人肺成纤维细胞(HFL1)的外切体预温育30min，3-甲基腺嘌呤(自噬抑制剂)和雷帕霉素(自噬诱导剂)分别孵育1h，然后再用顺铂处理8h。收集细胞进行细胞凋亡检测、酶联免疫吸附试验(ELISA)、Western印迹和实时定量聚合酶链式反应(qRT-PCR)。在体内，我们建立了顺铂诱导的大鼠急性肾损伤模型。在顺铂治疗0.5h前，将hucMSC-Ex或HFl1-Ex通过肾被膜注入肾脏。HucMSC-Ex前肾被膜下注射3-甲基腺嘌呤和雷帕霉素。顺铂注射后3d处死动物。观察肾功能、Luminex测定、肾小管细胞凋亡和增殖及自噬反应。HucMSC-Ex体外抑制顺铂诱导的肾小管上皮细胞线粒体凋亡和炎性细胞因子的分泌。HucMSC-Ex可上调NRK-52E细胞自噬标志蛋白LC3B和自噬相关基因ATG5、ATG7的表达。雷帕霉素可模拟hucMSC-Ex对顺铂诱导的肾损伤的保护作用，而这种作用可被自噬抑制剂3-甲基腺嘌呤所消除。我们的研究结果表明，hucMSC-Ex诱导的自噬激活能有效减轻顺铂的肾毒性。因此，hucMSC-Ex的前处理可能是提高顺铂疗效的一种新方法。本文的在线版本(doi：10.1186/s13287-0160463-4)包含补充材料，授权用户可以使用。
The administration of cisplatin is limited due to its nephrotoxic side effects, and prevention of this nephrotoxicity of cisplatin is difficult. Mesenchymal stem cell (MSC)-derived exosomes have been implicated as a novel therapeutic approach for tissue injury. In this study, we demonstrated that the pretreatment of human umbilical cord MSC-derived exosomes (hucMSC-Ex) can prevent the development of cisplatin-induced renal toxicity by activation of autophagy in vitro and in vivo. In vitro, rat renal tubular epithelial (NRK-52E) cells were pre-incubated with exosomes from hucMSC or HFL1 (human lung fibroblast cells; as control) for 30 min, and 3-methyladenine (an autophagic inhibitor) and rapamycin (an autophagic inducer) for 1 h before cisplatin treatment for 8 h, respectively. Cells were harvested for apoptosis assay, enzyme-linked immunosorbent assay (ELISA), Western blot, and quantitative real-time polymerase chain reaction (qRT-PCR). In vivo, we constructed cisplatin-induced acute kidney injury rat models. Prior to treatment with cisplatin for 0.5 h, hucMSC-Ex or HFL1-Ex were injected into the kidneys via the renal capsule. 3-methyladenine and rapamycin were injected under the kidney capsule before hucMSC-Ex. All animals were sacrificed at 3 days after cisplatin injection. Renal function, Luminex assay, tubular apoptosis and proliferation, and autophagy response were evaluated. hucMSC-Ex inhibited cisplatin-induced mitochondrial apoptosis and secretion of inflammatory cytokines in renal tubular epithelial cells in vitro. hucMSC-Ex increased the expression of the autophagic marker protein LC3B and the autophagy-related genes ATG5 and ATG7 in NRK-52E cells. Rapamycin mimicked the effects of hucMSC-Ex in protecting against cisplatin-induced renal injury, while the effects were abrogated by the autophagy inhibitor 3-methyladenine in the animals. Our findings indicate that the activation of autophagy induced by hucMSC-Ex can effectively relieve the nephrotoxicity of cisplatin. Therefore, pre-treatment of hucMSC-Ex may be a new method to improve the therapeutic effect of cisplatin. The online version of this article (doi:10.1186/s13287-016-0463-4) contains supplementary material, which is available to authorized users.