Song Shiqing, Lin Faliang, Zhu Pengyan, Wu Changyan, Zhao Shuling, Han Qiao, Li Xiaomei
Yantai Yuhuangding Hospital of Qingdao University Medical College, Yantai, Shandong, China.
Yantai Hospital of Traditional Chinese Medicine, Yantai, Shandong, China.
PLoS One. 2017 Sep 7;12(9):e0184348. doi: 10.1371/journal.pone.0184348. eCollection 2017.
Cerebral stroke is a leading cause of death and permanent disability. The current therapeutic outcome of ischemic stroke (>85% of all strokes) is very poor, thus novel therapeutic drug is urgently needed. In vitro cell model of ischemia was established by oxygen-glucose deprivation (OGD) and in vivo animal model of ischemia was established by middle cerebral artery occlusion (MCAO). The effects of Spatholobus suberctus Dunn extract (SSCE) on OGD-induced cell injury, MCAO-induced neural injury and miR-494 level were all evaluated. The possible target genes were virtually screened utilizing bioinformatics and verified by luciferase assay. Subsequently, the effects of abnormally expressed miR-494 on OGD-induced cell injury and target gene expression were determined. Additionally, whether SSCE affected target gene expression through modulation of miR-494 was studied. Finally, the effects of aberrantly expressed Sox8 on OGD-induced injury and signaling pathways were estimated. SSCE reduced OGD-induced cell injury and ameliorated MCAO-induced neuronal injury, along with down-regulation of miR-494. Then, OGD-induced cell injury was increased by miR-494 overexpression but decreased by miR-494 silence. Sox8 was a target gene of miR-494, and SSCE could up-regulate Sox8 expression via down-regulating miR-494. Afterwards, OGD-induced cell injury was proved to be increased by Sox8 inhibition but reduced by Sox8 overexpression. Finally, OGD-induced inhibition of PI3K/AKT/mTOR and MAPK pathways was further inhibited by Sox8 silence but activated by Sox8 overexpression. SSCE ameliorates ischemia-induced injury both in vitro and in vivo by miR-494-mediated modulation of Sox8, involving activations of PI3K/AKT/mTOR and MAPK pathways.
脑卒是死亡和永久性残疾的主要原因。目前缺血性脑卒中（占所有脑卒中的85%以上）的治疗效果非常差，因此迫切需要新型治疗药物。通过氧糖剥夺（OGD）建立体外缺血细胞模型，通过大脑中动脉闭塞（MCAO）建立体内缺血动物模型。评估了鸡血藤提取物（SSCE）对OGD诱导的细胞损伤、MCAO诱导的神经损伤和miR-494水平的影响。利用生物信息学虚拟筛选可能的靶基因，并通过荧光素酶测定进行验证。随后，确定异常表达的miR-494对OGD诱导的细胞损伤和靶基因表达的影响。此外，研究了SSCE是否通过调节miR-494影响靶基因表达。最后，评估异常表达的Sox8对OGD诱导的损伤和信号通路的影响。SSCE减少了OGD诱导的细胞损伤，改善了MCAO诱导的神经元损伤，同时下调了miR-494。然后，miR-494过表达增加了OGD诱导的细胞损伤，而miR-494沉默则降低了该损伤。Sox8是miR-494的靶基因，SSCE可通过下调miR-494上调Sox8表达。之后，证实Sox8抑制增加了OGD诱导的细胞损伤，而Sox8过表达则降低了该损伤。最后，Sox8沉默进一步抑制了OGD诱导的PI3K/AKT/mTOR和MAPK通路的抑制，但Sox8过表达激活了这些通路。SSCE通过miR-494介导的Sox8调节在体外和体内改善缺血诱导的损伤，涉及PI3K/AKT/mTOR和MAPK通路的激活。
