摘要
目的探讨丹参注射液(Salvia Miltiorrhiza Injection,SMN)对SD大鼠来源的神经干细胞分化细胞神经突起发生的影响。方法取1日龄大鼠神经干细胞原代培养后传代至第4代,经鉴定符合神经干细胞特征,然后加入不同浓度的SMN进行干预后,分别为SMN 10μg/ml、SMN 20μg/ml、SMN 50μg/ml对神经干细胞进行体外诱导。显微图像Olympus Cell Sens Dimension软件实时采集分析测定细胞图像,测量不同浓度丹参注射液对突起发生的影响;用免疫组织化学方法检测突起微管结构标志物βⅢ-tubulin微管蛋白的表达,采用Western blot法检测βⅢ-tubulin在神经突起中特异性蛋白的表达以及细胞外调节蛋白激酶ERK磷酸化程度。结果 SMN诱导后突起发生率随着浓度的增加而增加,呈现明显的浓度依赖性,较空白对照组分别增加3.6%(10μg/ml),18.5%(20μg/ml)和26.0%(50μg/ml),βⅢ-tubulin表达增加(P<0.05),调节蛋白激酶ERK磷酸化水平明显升高(P<0.05)。结论丹参注射液体外可以诱导神经干细胞来源的细胞突起发生,在突触建立过程中发挥重要作用,其作用机制可能与上调蛋白激酶ERK有关。
Objective To investigate the induction of neural protrusions from neural stem cells derived from Salvia Miltiorrhiza Injection in rats. Methods From the 1 day old SD rat neural stem cells cultured from generation to the fourth generation,of Salvia Miltiorrhiza Injection interfered with different concentrations of Olympus CellSens Dimension is the real-time analysis software under the microscope to determine cell image,measuring different concentrations of Salvia miltiorrhiza SMI on neurite occurred;marker expression of βⅢ-tubulin the microtubule structure was detected by immunohistochemical method and projection neurons,Tuj1 antibodies were detected by Western blot method （beta III-tubulin） specific protein expression in neurons,extracellular regulated ERK phosphorylation level. Results Neural stem cells after induced by SMI after projection incidence increases with increasing concentration,obvious concentration dependence,increased by 3.6% compared with the control group （10 μg/ml）tested,18.5% （20 μg/ml） of Salvia miltiorrhiza injection tested and 26% （40 μg/ml）;Salvia Miltiorrhiza Injection may promote βⅢ-tubulin （P〈0.05）,the expression of ERK increased significantly （P〈0.05）. Conclusion Salvia miltiorrhiza injection can induce the occurrence of cell protrusion,may take a crucial role in development of synapse. The mechanism of its action may be associated with up- regulation of protein kinase ERK.
作者
王光辉
钟鸣
张敏娜
WANG Guanghui;ZHONG Ming;ZHANG Minna(College of Pharmacy Jining Medical University,Rizhao 276826,Chin)
出处
《中风与神经疾病杂志》
CAS
2018年第7期580-584,共5页
Journal of Apoplexy and Nervous Diseases
基金
国家自然科学基金项目(61473043)
济宁市科技发展计划项目(201557127)
济宁医学院青年科学基金项目(JY2015KJ005)
济宁医学院教师科研扶持基金项目(JY2017KJ050)
