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首页> 美国卫生研究院文献>Journal of Cellular and Molecular Medicine >Overexpression of miR‐19a and miR‐20a in iPS‐MSCs preserves renal function of chronic kidney disease with acute ischaemia‐reperfusion injury in rat
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Overexpression of miR‐19a and miR‐20a in iPS‐MSCs preserves renal function of chronic kidney disease with acute ischaemia‐reperfusion injury in rat

机译：IPS-MSCs中miR-19a和miR-20a的过度表达在大鼠中保留了慢性肾疾病的肾功能急性肾脏血液再灌注损伤

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This study tested the hypothesis that therapy with double overexpression of miR‐19a‐3p and miR‐20a‐5p (miRDOE) to human inducible pluripotent stem cell–derived mesenchymal stem cells (iPS‐MSCs) was superior to iPS‐MSCs alone for preserving renal function in rat with pre‐existing chronic kidney disease (CKD), followed by ischaemia‐reperfusion (IR) injury. In vitro study demonstrated that the protein expressions of oxidative stress (NOX‐1/NOX‐2/NOX4/oxidized protein/p22phox), inflammatory downstream signalling (TLR2&4/MyD88/TRAF6/IKK‐ß/p‐NFκB/IL‐1ß/IL‐6/MMP‐9) and cell apoptosis/death signalling (cleaved caspase‐3/mitochondrial Bax/p‐ERKs/p‐JNK/p‐p38) at time‐points of 24‐hour/48‐hour cell cultures were significantly increased in p‐Cresol‐treated NRK‐52E cells than in the control that was significantly reversed by miR‐19a‐3p‐transfected iPS‐MSC (all P < .001). Animals were categorized into group 1 (sham‐operated control), group 2 (CKD‐IR), group 3 (CKD‐IR + oligo‐miRDOE of iPS‐MSCs/6.0 ×105/intra‐renal artery transfusion/3 hours after IR procedure), group 4 (CKD‐IR + iPS‐MSCs) and group 5 (CKD‐IR + miRDOE of iPS‐MSCs/6.0 ×105/intra‐renal artery transfusion/3 hour after IR procedure). By day 35, the creatinine/BUN levels were lowest in group 1, highest in group 2 and significantly lower in group 5 than in groups 3 and 4 (all P < .0001) but they showed no difference between the latter two groups. The protein expressions of oxidative stress, inflammatory downstream signalling and cell apoptosis/death signalling exhibited an identical pattern of creatinine level among the five groups (all P < .00001). Also, the microscopic findings demonstrated that the kidney injury score/fibrotic areaumber of inflammatory cells (CD14+/CD68+) exhibited an identical pattern of creatine level (all P < .0001). The miRDOE of iPS‐MSCs was superior to iPS‐MSCs for preserving the residual kidney function and architecture in CKD‐IR rat.
机译：该研究测试了用miR-19a-3p和miR-20a-5p（miRdoe）对人诱导多能干细胞衍生的间充质干细胞（ips-mscs）的双重过度表达治疗的假设优于IPS-MSCs以进行保留具有预先存在的慢性肾病（CKD）大鼠肾功能，其次是缺血再灌注（IR）损伤。体外研究表明，氧化应激的蛋白质表达（NOX-1 / NOX-2 / NOX4 /氧化蛋白/ p22phox），炎症下游信号（TLR2和4 / MYD88 / TRAF6 / IKK-β/ P-NFκB/ IL-1ß/在24小时/ 48小时细胞培养物的时间点，IL-6 / MMP-9）和细胞凋亡/死导（切割的Caspase-3 /线粒体Bax / P-Erks / P-P38）是在p折叠治疗的NRK-52e细胞中显着增加，比MIR-19A-3P转染的IPS-MSC显着逆转（所有P <.001）。将动物分为第1组（假手术控制），第2组（CKD-IR），第3组（IPS-MSCs / 6.0×105 /内部动脉内动脉输出/ 3小时后的CKD-IR + Oligo-MideOre / 3小时程序），第4组（CKD-IR + IPS-MSCs）和第5组（IPS-MSCs / 6.0×10 5 /内部动脉输出的CKD-IR + MIDOE / IR肾内动脉输出/ IR手术后3小时）。在第35天，在第1组中，群体/面包水平最低，第2组中最高，比第3组群体显着降低（所有P <.0001），但它们在后两组之间没有差异。氧化应激，炎症下游信号和细胞凋亡/死导/死导的蛋白质表达在五组（所有P <.00001）中表现出相同的肌酐水平模式。此外，微观研究结果表明，肾脏损伤得分/纤维化面积/炎性细胞（CD14 + / CD68 +）的数量表现出相同的肌酸水平模式（所有P <.0001）。 IPS-MSCs的MIDEOES优于IPS-MSCs，用于保留CKD-IR RAT中的残留肾功能和结构。

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