Yang Junhong, Gao Feng, Zhang Yunke, Liu Ying, Zhang Di
The First Department of Encephalopathy, The First Affiliated Hospital of Henan University of Traditional Chinese Medicine, Zhengzhou, 450000, China.
Department of Neuroimmunology, Henan Province Academy of Medical and Pharmaceutical, Zhengzhou University, Zhengzhou, 450052, China.
J Mol Neurosci. 2015 Aug;56(4):898-906. doi: 10.1007/s12031-015-0539-0. Epub 2015 Mar 22.
Buyang Huanwu decoction (BYHWD) has been used for centuries to treat paralysis and stroke. Previously, we have demonstrated that BYHWD combined with mesenchymal stem cell (MSC) transplantation attenuates ischemic injury partly by upregulating angiogenesis. However, the mechanisms of this drug for stroke treatment are not completely understood. Here, we aimed to clarify the mechanism of BYHWD on angiogenesis mediated by MSCs. Firstly, we verified microvessels with a size of 50-100 nm produced by either MSCs or MSCs treated by 500 μg/ml BYHWD. These exosomes were purified and found to be able to activate vascular endothelial growth factor (VEGF) expression in endothelial cells (ECs). Moreover, exosomes from MSCs and MSCs treated by BYHWD induced elevated microRNA (miRNA)-126 expression and reduced miR-221 and miR-222 expression. In MSCs, disruption of dicer, an enzyme responsible for miRNA maturation, by dicer small interfering RNA (siRNA), or RNase pretreatment abolished this ability of the exosomes. Additionally, exosomes from MSCs treated by BYHWD promoted VEGF and Ki-67 expression and augmented vascular density in rat brain after bilateral carotid artery ligation. In conclusion, our study revealed that BYHWD exposure augmented angiogenetic miRNA and VEGF expression in exosomes secreted by MSCs and elevated angiogenesis in rat brain.
补阳还五汤（BYHWD）已被用于治疗瘫痪和中风数百年。此前，我们已经证明，补阳还五汤联合间充质干细胞（MSC）移植可通过上调血管生成来部分减轻缺血性损伤。然而，这种药物治疗中风的机制尚未完全明确。在此，我们旨在阐明补阳还五汤对间充质干细胞介导的血管生成的作用机制。首先，我们验证了由间充质干细胞或经500μg/ml补阳还五汤处理的间充质干细胞产生的大小为50-100nm的微血管。这些外泌体被纯化，并发现能够激活内皮细胞（ECs）中血管内皮生长因子（VEGF）的表达。此外，来自间充质干细胞和经补阳还五汤处理的间充质干细胞的外泌体诱导了微小RNA（miRNA）-126表达的升高，并降低了miR-221和miR-222的表达。在间充质干细胞中，通过dicer小干扰RNA（siRNA）破坏负责miRNA成熟的酶dicer，或进行RNase预处理，消除了外泌体的这种能力。此外，经补阳还五汤处理的间充质干细胞的外泌体促进了双侧颈动脉结扎后大鼠脑中VEGF和Ki-67的表达，并增加了血管密度。总之，我们的研究表明，补阳还五汤处理增强了间充质干细胞分泌的外泌体中血管生成相关miRNA和VEGF的表达，并提高了大鼠脑中的血管生成。
