Exosomes secreted by human urine-derived stem cells could prevent kidney complications from type I diabetes in rats.
Exosomes secreted by human urine-derived stem cells could prevent kidney complications from type I diabetes in rats.
复制标题
人类尿液干细胞分泌的外泌体可以预防大鼠 I 型糖尿病的肾脏并发症
DOI：
10.1186/s13287-016-0287-2
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发表时间：
2016-02-06
影响因子：
7.5
通讯作者：
Wang NS
中科院分区：
文献类型：
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作者：
Jiang ZZ;Liu YM;Niu X;Yin JY;Hu B;Guo SC;Fan Y;Wang Y;Wang NS
糖尿病肾病是糖尿病患者最严重的并发症之一。目前，对于糖尿病肾病尚无令人满意的治疗方法。干细胞是目前开发糖尿病肾病新疗法的主要候选对象，因为它们可能主要通过旁分泌机制发挥治疗作用。据报道，源自干细胞的外泌体在肾损伤中起重要作用。在本文中，我们试图研究从尿干细胞中提取的外泌体在体内和体外是否能在早期预防糖尿病肾病。
采用超滤联合纯化方法从尿源性干细胞条件培养基中分离外泌体（USCs - Exo）。然后利用透射电子显微镜、可调电阻脉冲传感分析和蛋白质印迹法通过形态、大小和特异性生物标志物对USCs - Exo进行验证。建立链脲佐菌素诱导的Sprague - Dawley大鼠模型后，通过每周尾静脉注射USCs - Exo或对照物直至12周，观察USCs - Exo对肾损伤和血管生成的影响。在体外，用USCs - Exo处理在高糖培养基中培养的足细胞，以测试USCs - Exo对足细胞凋亡的保护作用。同时，通过酶联免疫吸附试验研究USCs - Exo和尿源性干细胞条件培养基中促进血管再生的潜在因子。
培养尿源性干细胞，并通过CD29、CD73、CD90和CD44抗原的阳性标志物以及CD34、CD45和HLA - DR的阴性标志物进行验证。USCs - Exo是直径约50 - 100nm的球形囊泡，特异性标志物包括CD9、CD63和CD81。在糖尿病大鼠中，静脉注射USCs - Exo可能减少尿量和尿微量白蛋白排泄，防止足细胞和肾小管上皮细胞凋亡，抑制caspase - 3过度表达并增加肾小球内皮细胞增殖。此外，USCs - Exo可减少体外高糖诱导的足细胞凋亡。USCs - Exo含有潜在因子，包括生长因子、转化生长因子 - β1、血管生成素和骨形态发生蛋白 - 7，这些因子可能与血管再生和细胞存活有关。
USCs - Exo可能具有通过抑制足细胞凋亡以及促进血管再生和细胞存活来预防糖尿病肾损伤的潜力。
本文的在线版本（doi:10.1186/s13287 - 016 - 0287 - 2）包含补充材料，授权用户可获取。
BackgroundDiabetic nephropathy is one of the most serious complications in patients with diabetes. At present, there are no satisfactory treatments available for diabetic nephropathy. Stem cells are currently the main candidates for the development of new treatments for diabetic nephropathy, as they may exert their therapeutic effects mainly through paracrine mechanisms. Exosomes derived from stem cells have been reported to play an important role in kidney injury. In this article, we try to investigate whether exosomes retrieved from urine stem cells could itself prevent diabetic nephropathy at an early stage in vivo and in vitro.MethodsExosomes from conditioned medium of urine-derived stem cells (USCs-Exo) were isolated using ultrafiltration-combined purification methods. USCs-Exo were then verified by morphology, size, and specific biomarkers using transmission electron microscopy, tunable resistive pulse sensing analysis, and western blotting. After establishment of the streptozotocin-induced Sprague–Dawley rat model, the effects of USCs-Exo on kidney injury and angiogenesis were observed via weekly tail intravenous injection of USCs-Exo or control until 12 weeks. In vitro, podocytes cultured in high-glucose medium were treated with USCs-Exo to test the protective effect of USCs-Exo on podocytic apoptosis. Meanwhile, the potential factors in promoting vascular regeneration in USCs-Exo and urine-derived stem cell conditioned medium were investigated by enzyme-linked immunosorbent assay.ResultsUrine-derived stem cells were cultured and were verified by positive markers for CD29, CD73, CD90 and CD44 antigens, and negative markers for CD34, CD45 and HLA-DR. USCs-Exo were approximately 50–100 nm spherical vesicles, and the specific markers included CD9, CD63 and CD81. Intravenous injections of USCs-Exo could potentially reduce the urine volume and urinary microalbumin excretion, prevent podocyte and tubular epithelial cell apoptosis, suppress the caspase-3 overexpression and increase glomerular endothelial cell proliferation in diabetic rats. In addition, USCs-Exo could reduce podocytic apoptosis induced by high glucose in vitro. USCs-Exo contained the potential factors, including growth factor, transforming growth factor-β1, angiogenin and bone morphogenetic protein-7, which may be related with vascular regeneration and cell survival.ConclusionUSCs-Exo may have the potential to prevent kidney injury from diabetes by inhibiting podocyte apoptosis and promoting vascular regeneration and cell survival.