诱导多能干细胞源性间充质干细胞中 miR-19a 和 miR-20a 的双重过表达可有效保护扩张型心肌病大鼠的左心室功能。
Double overexpression of miR-19a and miR-20a in induced pluripotent stem cell-derived mesenchymal stem cells effectively preserves the left ventricular function in dilated cardiomyopathic rat.
机构信息
Division of Thoracic and Cardiovascular Surgery, Department of Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, 83301, Taiwan.
Division of Cardiology, Department of Internal Medicine, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University, College of Medicine, 123, Dapi Road, Niaosung Dist, Kaohsiung, 83301, Taiwan.
出版信息
Stem Cell Res Ther. 2021 Jun 29;12(1):371. doi: 10.1186/s13287-021-02440-4.
BACKGROUND
This study tested the hypothesis that double overexpression of miR-19a and miR-20a (dOex-mIRs) in human induced pluripotent stem cell (iPS)-derived mesenchymal stem cells (MSCs) effectively preserved left ventricular ejection fraction (LVEF) in dilated cardiomyopathy (DCM) (i.e., induced by doxorubicin) rat.
METHODS AND RESULTS
In vitro study was categorized into groups G1 (iPS-MSC), G2 (iPS-MSC), G3 (iPS-MSC + HO/100uM), and G4 (iPS-MSC + HO/100uM). The in vitro results showed the cell viability was significantly lower in G3 than in G1 and G2, and that was reversed in G4 but it showed no difference between G1/G2 at time points of 6 h/24 h/48 h, whereas the flow cytometry of intra-cellular/mitochondrial oxidative stress (DCFA/mitoSOX) and protein expressions of mitochondrial-damaged (cytosolic-cytochrome-C/DRP1/Cyclophilin-D), oxidative-stress (NOX-1/NOX2), apoptotic (cleaved-caspase-3/PARP), fibrotic (p-Smad3/TGF-ß), and autophagic (ratio of LC3B-II/LC3BI) biomarkers exhibited an opposite pattern of cell-proliferation rate (all p< 0.001). Adult-male SD rats (n=32) were equally divided into groups 1 (sham-operated control), 2 (DCM), 3 (DCM + iPS-MSCs/1.2 × 10 cells/administered by post-28 day's DCM induction), and 4 (DCM + iPS-MSC/1.2 × 10 cells/administered by post-28 day's DCM induction) and euthanized by day 60 after DCM induction. LV myocardium protein expressions of oxidative-stress signaling (p22-phox/NOX-1/NOX-2/ASK1/p-MMK4,7/p-JNK1,2/p-cJUN), upstream (TLR-4/MAL/MyD88/TRIF/TRAM/ TFRA6/IKK/NF-κB) and downstream (TNF-α/IL-1ß/MMP-9) inflammatory signalings, apoptotic (cleaved-PARP/mitochondrial-Bax), fibrotic (Smad3/TGF-ß), mitochondrial-damaged (cytosolic-cytochrome-C/DRP1/cyclophilin-D), and autophagic (beclin1/Atg5) biomarkers were highest in group 2, lowest in group 1 and significantly lower in group 4 than in group 3, whereas the LVEF exhibited an opposite pattern of oxidative stress (all p< 0.0001).
CONCLUSION
iPS-MSC therapy was superior to iPS-MSC therapy for preserving LV function in DCM rat.
背景
本研究旨在检验 miR-19a 和 miR-20a 双重过表达（dOex-mIRs）在人诱导多能干细胞（iPS）衍生的间充质干细胞（MSCs）中是否能有效保留阿霉素诱导的扩张型心肌病（DCM）大鼠的左心室射血分数（LVEF）。
方法和结果
体外研究分为 G1（iPS-MSC）、G2（iPS-MSC）、G3（iPS-MSC+HO/100μM）和 G4（iPS-MSC+HO/100μM）组。体外结果显示，G3 组细胞活力明显低于 G1 组和 G2 组，而 G4 组则逆转，但在 6 h/24 h/48 h 时，G1/G2 之间无差异，而细胞内/线粒体氧化应激（DCFA/mitoSOX）的流式细胞术和线粒体损伤（胞质细胞色素-C/DRP1/环胞素-D）、氧化应激（NOX-1/NOX2）、凋亡（cleaved-caspase-3/PARP）、纤维化（p-Smad3/TGF-β）和自噬（LC3B-II/LC3BI 比值）生物标志物的蛋白表达呈现出与细胞增殖率相反的模式（均 p<0.001）。成年雄性 SD 大鼠（n=32）等分为 4 组：1 组（假手术对照）、2 组（DCM）、3 组（DCM+iPS-MSCs/1.2×106 个细胞/在 DCM 诱导后 28 天给药）和 4 组（DCM+iPS-MSCs/1.2×106 个细胞/在 DCM 诱导后 28 天给药），并在 DCM 诱导后 60 天处死。LV 心肌氧化应激信号（p22-phox/NOX-1/NOX-2/ASK1/p-MMK4,7/p-JNK1,2/p-cJUN）、上游（TLR-4/MAL/MyD88/TRIF/TRAM/TFRA6/IKK/NF-κB）和下游（TNF-α/IL-1β/MMP-9）炎症信号、凋亡（cleaved-PARP/线粒体-Bax）、纤维化（Smad3/TGF-β）、线粒体损伤（胞质细胞色素-C/DRP1/环胞素-D）和自噬（beclin1/Atg5）生物标志物在 2 组中最高，在 1 组中最低，在 4 组中明显低于 3 组，而左心室射血分数（LVEF）则呈现出与氧化应激相反的模式（均 p<0.0001）。
结论
iPS-MSC 治疗优于 iPS-MSC 治疗，可保留 DCM 大鼠的 LV 功能。
Zotero 插件