主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

北京中医药大学学报 ›› 2017, Vol. 40 ›› Issue (6): 483-491.doi: 10.3969/j.issn.1006-2157.2017.06.007

• 科技之窗 • 上一篇    下一篇

脑络欣通对大鼠脑缺血再灌注后海马mRNA的特异性表达及Wnt/β-catenin的影响

邓勇, 王键#, 谭辉, 何玲   

  1. 安徽中医药大学 安徽 230038
  • 收稿日期:2016-12-20 出版日期:2017-06-10 发布日期:2017-06-10
  • 通讯作者: 王键,男,教授,博士生导师,主要研究方向:新安医学及其治则治法研究,E-mail:wangjian6301@163.com
  • 作者简介:邓勇,男,硕士,副研究馆员,在读博士生
  • 基金资助:
    国家中医药管理局“新安王氏内科流派传承工作室”建设项目(No.国中医药人教函{2012}228号),国家自然科学基金青年项目(No.81603523),安徽省高校自然科学研究重点项目(No.KJ2016A404)

Effects of Naoluo Xintong on mRNA, wnt/β-catenin in hippocampus of MCAO/R rats*

DENG Yong, WANG Jian#, TAN Hui, HE Ling   

  1. Anhui University of Traditional Chinese Medicine,Anhui, 230038, China
  • Received:2016-12-20 Online:2017-06-10 Published:2017-06-10

摘要: 目的 研究脑络欣通对大鼠脑缺血再灌注损伤神经保护的作用机制。 方法 雄性SD大鼠144只,随机分为假手术组、模型组、对照组、治疗组,每组36只,各组随机数字表法分成1、3、7 d组,每个时间节点12只。复制脑缺血再灌注(MCAO/R)大鼠模型。治疗组造模后分别于1、3、7 d给予脑络欣通,对照组造模后分别1、3、7 d给予通心络。脑络欣通配制成浓度为0.26 g/mL的溶液,通心络配制成浓度为0.02 g/mL的溶液,均按10 mL/(kgd)灌胃给药。假手术组及模型组分别1、3、7 d予以等量生理盐水灌胃。采用激光多普勒血流仪(LDF)测定大鼠缺血侧局部脑血流量(rCBF)。解剖大鼠取出脑组织并分离海马,采用HE染色法检测大鼠海马DG区;采用实时荧光定量PCR技术(RT-PCR)测定miRNA9a、miRNA15a、miRNA124的表达;采用Western blot法检测Wnt3a、Wnt5a、β-catenin的表达。 结果 与假手术组比较,模型组大鼠脑组织明显缺血,脑血流量显著降低(P<0.01),海马中miRNA9a、miRNA15a表达均显著降低(P<0.01), miRNA124显著增高(P<0.01),Wnt3a、Wnt5a、β-catenin表达显著增高(P<0.01);与模型组比较,脑络欣通1、3、7 d组大鼠脑血流量恢复明显,海马中miRNA9a、miRNA15a表达显著增高(P<0.01),miRNA124表达水平显著降低(P<0.01),Wnt3a、Wnt5a、β-catenin表达显著增高(P<0.01)。 结论 脑络欣通对于MCAO/R大鼠脑损伤有明显疗效,可通过提高局部脑血流量,提高海马miRNA9a、miRNA15a表达,降低海马miRNA124表达,提高Wnt3a、Wnt5a、β-catenin的表达,起到抑制缺血再灌注对大鼠脑组织损伤的作用。

关键词: 脑络欣通, 益气活血通络法, 脑缺血再灌注, mRNA, Wnt/β-catenin, 大鼠

Abstract: Objective To investigate the mechanism of protective effects of Naoluo Xintong (NLXT) on cerebral ischemia-reperfusion injury in rats. Methods 144 male SD rats were randomly divided into 4 groups (n = 36 in each group): sham-operation group, model group, normal control group and treatment group. Each group was randomly further divided into 1/3/7d subgroups (n=12 in each subgroup). Treatment group was administered NLXT after modeling on the 1st, 3rd, and 7th days, and normal control group was given Tongxinluo on the 1st, 3rd, and 7th days. The treatment group and control group was given 10 mL/(kgd) of NLXT liquid (concentration = 0.26 g/mL) and Tongxinluo liquid (concentration = 0.02 g/mL) respectively. The model group and sham-operation group were orally gavaged with isopyknic physiological saline solution on the 1st, 3rd, 7th days. Regional cerebral blood flow (rCBF) of cerebral infarction side was measured through laser Doppler Flowmetry (LDF). All the rats were dissected and hippocampal tissues were removed from their brains. Then DG area of rat’s hippocampus was measured by using HE staining. MiRNA9a, miRNA15a, and miRNA124 in the hippocampus were detected with RT-PCR; Wnt3a, Wnt5a and β-catenin was detected by using Western blot method. Results Compared with the sham-operation group, rats of the model group presented apparent cerebral ischemia; levels of miRNA124, Wnt3a, Wnt5a, and β-catenin expression in hippocampal area significantly increased (P<0.01; Levels of miRNA9a and miRNA15 expression markedly decreased. Compared with the model group, the blood flow in brain increased significantly on the 1st, 3rd, and 7th days (P<0.01; Levels of miRNA9a, miRNA15a, Wnt3a, Wnt5a, and β-catenin expression in hippocampal area increased significantly; miRNA124 expression decreased significantly. Conclusion NLXT had a good effect of treating cerebral injury of MCAO/R rats. This medicine could increase the cerebral blood flow, miRNA9a, miRNA15a, Wnt3a, Wnt5a, and β-catenin expressions, and decrease miRNA124 expression in rat hippocampus, i.e. inhibiting ischemia-reperfusion cerebral impairment in rats.

Key words: Naoluo Xintong granules, supplementing qi and nourishing blood to regulate meridians method, ischemia reperfusion injury, mRNA, wnt/β-catenin, rats

中图分类号: 

  • R285.5