Protective effect of miRNA-containing extracellular vesicles derived from mesenchymal stromal cells of old rats on renal function in chronic kidney disease
Protective effect of miRNA-containing extracellular vesicles derived from mesenchymal stromal cells of old rats on renal function in chronic kidney disease
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含miRNA的老年大鼠间充质基质细胞外囊泡对慢性肾病肾功能的保护作用
DOI：
10.1186/s13287-020-01792-7
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发表时间：
2020-07-08
影响因子：
7.5
通讯作者：
Shi, Yun Cong
中科院分区：
文献类型：
--
作者：
Wang, Yan;Guo, Yi Fang;Shi, Yun Cong
间充质基质细胞（MSCs）在预防细胞和组织纤维化中起着重要作用。在组织和器官损伤恢复过程中，衰老可能会降低间充质干细胞的功能。间充质干细胞释放的细胞外囊泡（EVs）有助于肾损伤的修复。我们探讨了衰老对来自MSCs的ev （msc - ev）的影响，并检测了来自老年大鼠的表达低水平miR-294/miR-133的msc - ev对慢性肾脏疾病（CKD）的保护作用。方法采用单侧输尿管梗阻（UUO）模型，观察3月龄和18月龄雄性Fisher 344大鼠骨髓间充质干细胞- ev对肾脏纤维化的影响。在UUO前将pLV-miR-294/pLV-miR-133模拟物/抑制剂注射到年轻大鼠和老年大鼠体内，检测miR-294/miR-133对CKD肾功能的影响，miR-294/miR-133在msc - ev和老年大鼠血清中降低。采用转化生长因子- β 1 （tgf - β 1）诱导的人肾近端小管上皮细胞（HK2）体外模拟肾纤维化病理过程。Western blotting检测HK2细胞上皮/间充质标记物的表达和磷酸化蛋白的表达。结果mscs - ev对老年大鼠uuo诱导的CKD的抑制作用弱于年轻大鼠。在老年大鼠骨髓间质干细胞和血清中下调mirna （miR-294和miR-133）可明显减轻uuo诱导的老年大鼠肾损伤。miR-294和miR-133过表达减轻了tgf - β 1介导的HK2细胞上皮间质转化（EMT），并且在miR-294和miR-133过表达的HK2细胞中，tgf - β 1诱导的SMAD2/3和ERK1/2磷酸化的明显增加被阻止。结论msc - ev抑制肾纤维化的能力随年龄的增长而下降。msc - ev和血清中的miR-294/miR-133对老年大鼠肾纤维化和HK2细胞的EMT有重要影响。此外，在tgf - β 1介导的EMT过程中，miR-294/miR-133过表达阻止了HK2细胞中SMAD2/3和ERK1/2的磷酸化。这些发现表明，miR-294/miR-133可能对老年人肾纤维化和相关肾功能障碍具有治疗作用。
Introduction Mesenchymal stromal cells (MSCs) play an important role in the prevention of cell and tissue fibrosis. Senescence may decrease the function of MSCs during recovery from tissue and organ damage. Extracellular vesicles (EVs) released from MSCs contribute to the repair of kidney injury. We explored the influence of senescence on EVs derived from MSCs (MSC-EVs) and detected the protective effects of MSC-EVs expressing low levels of miR-294/miR-133 derived from old rats against chronic kidney disease (CKD). Methods The effects of MSC-EVs derived from 3-month-old and 18-month-old male Fisher 344 rats on renal fibrosis were explored in a unilateral ureteral obstruction (UUO) model. pLV-miR-294/pLV-miR-133 mimic/inhibitor were injected into young and old rats before UUO to detect the effects of miR-294/miR-133, which were decreased in MSC-EVs and sera from old rats, on renal function in CKD. Transforming growth factor-beta 1 (TGF-beta 1)-induced human renal proximal tubular epithelial (HK2) cells were used to imitate the pathological process of renal fibrosis in vitro. Western blotting was used to assess the expression of epithelial/mesenchymal markers and phosphorylation of proteins in HK2 cells. Results The inhibition of UUO-induced CKD by MSC-EVs was weaker in old rats than in young rats. Downregulation of miRNAs (miR-294 and miR-133) in both MSC-EVs and sera from old rats obviously attenuated UUO-induced renal injury in old rats. miR-294 and miR-133 overexpression mitigated TGF-beta 1-mediated epithelial-mesenchymal transition (EMT) in HK2 cells, and the obvious increase in the phosphorylation of both SMAD2/3 and ERK1/2 induced by TGF-beta 1 was prevented in miR-294- and miR-133-overexpressing HK2 cells. Conclusions The ability of MSC-EVs to inhibit renal fibrosis decreased with age. miR-294/miR-133 in MSC-EVs and sera had an important effect on renal fibrosis in old rats and on EMT in HK2 cells. Furthermore, miR-294/miR-133 overexpression prevented SMAD2/3 and ERK1/2 phosphorylation in HK2 cells during TGF-beta 1-mediated EMT. These findings show that miR-294/miR-133 may be therapeutic in renal fibrosis and related renal dysfunction in elderly individuals.