Mesenchymal stem cells–microvesicle-miR-451a ameliorate early diabetic kidney injury by negative regulation of P15 and P19
Mesenchymal stem cells–microvesicle-miR-451a ameliorate early diabetic kidney injury by negative regulation of P15 and P19
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间充质干细胞微泡-miR-451a通过负调节P15和P19改善早期糖尿病肾损伤
DOI：
10.1177/1535370218819726
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发表时间：
2018-11
影响因子：
3.2
通讯作者：
Lu Yanrong
中科院分区：
文献类型：
--
作者：
Zhong Lin;Liao Guangneng;Wang Xiaojiao;Li Lan;Zhang Jie;Chen Younan;Liu Jingping;Liu Shuyun;Wei Lingling;Zhang Wengen;Lu Yanrong
骨髓间充质干细胞（mesenchymalstemcells，MSCs）微泡（Microvesicles，MV）是近年来发现的一种细胞间通讯的新途径。本研究旨在探讨MVs穿梭的miR-451 a对糖尿病肾病（DN）伴高尿酸血症患者肾纤维化和上皮间质转化（EMT）的影响及其机制。采用梯度离心法从MSC培养液中分离MVs。通过qPCR分析MSC和MV中miR-451 a的水平。在高糖高尿酸血症诱导的细胞（HK-2）和小鼠模型中检测miR-451 a、E-cadherin、α-SMA、P15 INK 4 b（P15）和P19 INK 4d（P19）的变化。流式细胞仪分析细胞周期的变化。BrdU法检测细胞增殖能力，CCK 8法检测细胞活力。进行双荧光素酶报告基因测定以确定靶结合位点。观察小鼠肾功能及肾组织学变化。MV显示与MSC相同的表面标志物，但miR-451 a表达高得多（比MSC高4.87 ± 2.03倍）。miR-451 a在损伤的HK-2细胞和肾脏中分别下降至26% ± 11%和6.7% ± 0.82%。MV-miR-451 a在体外可促进HK 2细胞增殖和存活，在体内可减轻肾脏形态和功能损伤。此外，MV-miR-451 a的输注降低了α-SMA的水平并提高了E-cadherin的表达。这些作用是通过miR-451 a靶向P15和P19的3′-UTR位点来改善细胞周期阻滞和下调P15和P19的作用。本研究表明，MSC-MV-miR-451 a在体内外均能抑制细胞周期抑制因子P15和P19，使细胞周期阻滞重新启动，逆转EMT，因此miR-451 a有望成为DN治疗的新靶点。影响声明MSCs修复糖尿病肾病损伤肾脏的机制尚不清楚。在研究中，MV显示出与MSC相同的表面标记，但MiR-451 a表达要高得多。miR-451 a在受损的HK-2细胞和肾脏中均减少。MV-miR-451 a在体外刺激细胞增殖和活力，并在体内促进损伤肾脏的结构和功能改善。输注MV-miR-451 a通过减少α-SMA和增加E-钙粘蛋白改善EMT。这些作用依赖于改善细胞周期阻滞以及通过miR-451 a结合其3′-UTR区域下调P15和P19。本研究表明，MSC-MV-miR-451 a在体内和体外均能特异性抑制细胞周期抑制剂，使阻滞的细胞周期重新启动，逆转EMT。因此，miR-451 a可能成为DN治疗的新靶点。
Microvesicles (MVs) from mesenchymal stem cells (MSCs) have been reported as a new communicated way between cells. This study evaluated the influence and underlying mechanism of MVs-shuttled miR-451a on renal fibrosis and epithelial mesenchymal transformation (EMT) in diabetic nephropathy (DN) with hyperuricemia. MVs were isolated from MSCs-cultured medium by gradient ultracentrifugation. The level of miR-451a in MSCs and MVs was analyzed by qPCR. The changes of miR-451a, E-cadherin, α-SMA, P15INK4b (P15), and P19INK4d (P19) were measured in hyperglycosis and hyperuricemia-induced cell (HK-2) and mouse models. The changes of cell cycle were analyzed by flow cytometry. The ability of proliferation and viability was measured by BrdU and CCK8, respectively. Dual-luciferase reporter assays were conducted to determine the target binding sites. The renal function and histological changes of mice were analyzed. MVs showed the same surface markers as MSCs but much higher miR-451a expression (4.87 ± 2.03 fold higher than MSCs). miR-451a was decreased to 26% ± 11% and 6.7% ± 0.82% in injured HK-2 cells and kidney, respectively. MV-miR-451a enhanced the HK2 cells proliferation and viability in vitro, and decreased the morphologic and functional injury of kidney in vivo. Moreover, infusion of MV-miR-451a reduced the level of α-SMA and raised E-cadherin expression. These effects were responsible for the improved arrested cell cycle and down-regulation of P15 and P19 via miR-451a targeting their 3′-UTR sites. This study demonstrated that MSC–MV-miR-451a could inhibit cell cycle inhibitors P15 and P19 to restart the blocked cell cycle and reverse EMT in vivo and in vitro, and thus miR-451a is potentially a new target for DN therapy. Impact statement The mechanism of MSCs repairing the injured kidney in diabetic nephropathy is not yet clear. In the research, MVs showed the same surface markers as MSCs but much higher MiR-451a expression. miR-451a was decreased in both injured HK-2 cells and kidneys. MV-miR-451a stimulated the cell proliferation and viability in vitro and promoted structural and functional improvements of injured kidney in vivo. Infusion of MV-miR-451a ameliorated EMT by reducing α-SMA and increasing E-cadherin. These effects relied on the improved cell cycle arrest and the down-regulation of P15 and P19 via miR-451a binding to their 3′-UTR region. This study demonstrated that MSC–MV-miR-451a could specifically inhibit cell cycle inhibitors to restart the blocked cell cycle and reverse EMT in vivo and in vitro. Therefore, miR-451a may be a new target for DN therapy.
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影响因子：
3.7
作者：
Collino F;Deregibus MC;Bruno S;Sterpone L;Aghemo G;Viltono L;Tetta C;Camussi G
通讯作者：
Camussi G
影响因子：
--
作者：
S. Yamagishi;T. Matsui
通讯作者：
S. Yamagishi;T. Matsui
影响因子：
2.2
作者：
Aiqun Zhu;T. Zou;Guangzhong Xiong;Jingping Zhang
通讯作者：
Aiqun Zhu;T. Zou;Guangzhong Xiong;Jingping Zhang
影响因子：
--
作者：
P. Romagnani
通讯作者：
P. Romagnani
DOI：
10.1073/pnas.0608249103
发表时间：
2006-11-14
影响因子：
11.1
作者：
Lee, Ryang Hwa;Seo, Min Jeong;Prockop, Darwin J.
通讯作者：
Prockop, Darwin J.