Chen Kuan-Hung, Chen Chih-Hung, Wallace Christopher Glenn, Yuen Chun-Man, Kao Gour-Shenq, Chen Yi-Ling, Shao Pei-Lin, Chen Yung-Lung, Chai Han-Tan, Lin Kun-Chen, Liu Chu-Feng, Chang Hsueh-Wen, Lee Mel S, Yip Hon-Kan
Department of Anesthesiology, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, Taiwan.
Department of Biological Sciences, National Sun Yat-Sen University, Kaohsiung, Taiwan.
Oncotarget. 2016 Nov 15;7(46):74537-74556. doi: 10.18632/oncotarget.12902.
We tested the hypothesis that combined xenogenic (from mini-pig) adipose-derived mesenchymal stem cell (ADMSC) and ADMSC-derived exosome therapy could reduce brain-infarct zone (BIZ) and enhance neurological recovery in rat after acute ischemic stroke (AIS) induced by 50-min left middle cerebral artery occlusion. Adult-male Sprague-Dawley rats (n = 60) were divided equally into group 1 (sham-control), group 2 (AIS), group 3 [AIS-ADMSC (1.2×106 cells)], group 4 [AIS-exosome (100μg)], and group 5 (AIS-exosome-ADMSC). All therapies were provided intravenously at 3h after AIS procedure. BIZ determined by histopathology (by day-60) and brain MRI (by day-28) were highest in group 2, lowest in group 1, higher in groups 3 and 4 than in group 5, but they showed no difference between groups 3 and 4 (all p < 0.0001). By day-28, sensorimotor functional results exhibited an opposite pattern to BIZ among the five groups (p < 0.005). Protein expressions of inflammatory (inducible nitric oxide synthase/tumor necrosis factor-α/nuclear factor-κB/interleukin-1β/matrix metalloproteinase-9/plasminogen activator inhibitor-1/RANTES), oxidative-stress (NOX-1/NOX-2/oxidized protein), apoptotic (caspase-3/ Poly-ADP-ribose polymerase), and fibrotic (Smad3/transforming growth factor-β) biomarkers, and cellular expressions of brain-damaged (γ-H2AX+/ XRCC1-CD90+/p53BP1-CD90+), inflammatory (CD11+/CD68+/glial fibrillary acid protein+) and brain-edema (aquaporin-4+) markers showed a similar pattern of BIZ among the groups (all n < 0.0001). In conclusion, xenogenic ADMSC/ADMSC-derived exosome therapy was safe and offered the additional benefit of reducing BIZ and improving neurological function in rat AIS.
联合异种(来自小型猪)脂肪间充质干细胞(ADMSC)和ADMSC来源的外泌体疗法可减少大鼠急性缺血性卒中(AIS)后梗死灶面积(BIZ)并促进神经功能恢复。AIS通过左侧大脑中动脉闭塞50分钟诱导。将成年雄性Sprague-Dawley大鼠(n = 60)平均分为1组(假手术对照组)、2组(AIS组)、3组[AIS-ADMSC组(1.2×106个细胞)]、4组[AIS-外泌体组(100μg)]和5组(AIS-外泌体-ADMSC组)。所有治疗均在AIS手术3小时后静脉给予。通过组织病理学(第60天)和脑MRI(第28天)测定的BIZ在2组中最高,在1组中最低,3组和4组高于5组,但3组和4组之间无差异(所有p < 0.0001)。到第28天,感觉运动功能结果在五组中与BIZ呈现相反模式(p < 0.005)。炎症(诱导型一氧化氮合酶/肿瘤坏死因子-α/核因子-κB/白细胞介素-1β/基质金属蛋白酶-9/纤溶酶原激活物抑制剂-1/调节激活正常T细胞表达和分泌因子)、氧化应激(NOX-1/NOX-2/氧化蛋白)、凋亡(半胱天冬酶-3/聚ADP核糖聚合酶)和纤维化(Smad3/转化生长因子-β)生物标志物的蛋白表达,以及脑损伤(γ-H2AX+/XRCC1-CD90+/p53BP1-CD90+)、炎症(CD11+/CD68+/胶质纤维酸性蛋白+)和脑水肿(水通道蛋白-4+)标志物的细胞表达在各组中呈现与BIZ相似的模式(所有n < 0.0001)。总之,异种ADMSC/ADMSC来源的外泌体疗法是安全的,并且在大鼠AIS中具有减少BIZ和改善神经功能的额外益处。