| 袁 梅，汤永红，袁海军，周成芳.miR-335在急性缺血性脑卒中表达下调及对钙调蛋白的调控作用.[J].中南医学科学杂志.,2015,43(3):276-280. |
| miR-335在急性缺血性脑卒中表达下调及对钙调蛋白的调控作用 |
| Negative Regulation of Calmodulin by miR-335 in Acute Ischemic Stroke |
| 投稿时间：2014-10-20 |
| DOI： |
| 中文关键词: 微小RNA-335 钙调蛋白 急性缺血性脑卒中 |
| 英文关键词:microRNA-335 calmodulin acute ischemic stroke |
| 基金项目:衡阳市科技计划项目（2014KJ40）. |
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| 中文摘要: |
| 目的 检测微小RNA-335在急性缺血性脑卒中（AIS）患者血清中的表达，并探讨其对钙调蛋白（CaM）调控的作用机制。 方法 采用实时荧光定量聚合酶链反应（qRT-PCR）法测定106例AIS患者及98例健康对照组血清中miR-335表达情况；用生物信息学数据库查询分析miR-335对钙调蛋白编码基因CALM1的靶向结合关系；脂质体转染法将miR-335模拟物、抑制物以及相应阴性对照物分别转染至人脐静脉内皮细胞（HUVECs）中并进行细胞培养，采用逆转录PCR（RT-PCR）和Western blot分别检测干预后HUVECs中钙调蛋白mRNA和蛋白的表达水平。 结果 与健康对照组相比较，AIS组患者血清miR-335表达明显下调（P<0.01）；生物信息学软件的查询分析显示CALM1存在miR-335潜在靶向结合位点；RT-PCR和Western blot检测结果显示miR-335模拟物组CaM mRNA和蛋白的表达水平较对照组显著下降（P<0.01），而miR-335抑制物组CaM mRNA和蛋白的表达水平较对照组显著增高（均P<0.05）。 结论 急性缺血性脑卒中患者血清miR-335表达明显下降，表达下调的miR-335可能通过上调钙调蛋白的表达从而发挥重要作用。 |
| 英文摘要: |
| Objective The miRNA dysfunction is suspected to be a contributing factor for many CNS pathologies including ischemic stroke.The present study investigates the expression of serum miR-335 in acute ischemic stroke （AIS）,and explores the underlying mechanisms of miR-335 in acute ischemic stroke （AIS）. Methods Blood samples were obtained from AIS patients （n106） and healthy controls （n 98）.MiR-335 was measured by using a quantitative real-time PCR.Bioinformatics database assay was used to determine whether calmodulin gene （CALM1） was the direct target of miR-335.The miR-335 mimics and inhibitors were transfected into human umbilical vein epithelial cells （HUVECs） with lipofectamine respectively.Calmodulin expression was determined by Western blot and reverse transcription-PCR （RT-PCR） in the cultured HUVECs. Results Compared with the healthy control group,the expression of serum miR-335 was significantly lower in patients with AIS group （P<0.01）.In addition,bioinformatics database assay show that the CALM1 present the binding sites,which was potentially targeted by miR-335.Finally，Transfection of the miR-335 mimic inhibited the expression of CaM mRNA and protein in the HUVEC compared to the control group （P<0.01）,while transfection of the miR-335 inhibitors increased the expression of CaM mRNA and protein （P<0.05）. Conclusion MiR-335 down-regulation may increase the ischemic brain injury through up-regulating the expression of CaM. |
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